Author’s Accepted Manuscript

A REVIEW ON SPECTROSCOPIC METHODS

FOR DETERMINATION OF NITRITE AND
NITRATE IN ENVIRONMENTAL SAMPLES

Priyanka Singh, Younus Raza Beg, Gokul Ram

Nishad
www.elsevier.com/locate/talanta

PII: S0039-9140(18)30839-7
DOI: https://doi.org/10.1016/j.talanta.2018.08.028
Reference: TAL18949
To appear in: Talanta
Received date: 18 June 2018
Revised date: 6 August 2018
Accepted date: 7 August 2018
Cite this article as: Priyanka Singh, Younus Raza Beg and Gokul Ram Nishad, A
REVIEW ON SPECTROSCOPIC METHODS FOR DETERMINATION OF
NITRITE AND NITRATE IN ENVIRONMENTAL SAMPLES, Talanta,
https://doi.org/10.1016/j.talanta.2018.08.028
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 A REVIEW ON SPECTROSCOPIC METHODS FOR DETERMINATION
OF NITRITE AND NITRATE IN ENVIRONMENTAL SAMPLES

Priyanka Singh*, Younus Raza Beg, Gokul Ram Nishad

Department of Chemistry, Govt. Digvijay PG Autonomous College, Rajnandgaon, 491441,

Chhattisgarh, India

*
Corresponding author. Assistant Professor Dr. Priyanka Singh Department of Chemistry,

Govt. Digvijay PG Autonomous College, Rajnandgaon, Chhattisgarh, India, PIN – 491441.

priyankasingh121@yahoo.com

ABSTRACT

Nitrate is an important pollutant found in environmental samples. Nitrate and nitrite pose various
environmental as well as health hazards. Different methods of determining nitrate in various
environmental samples developed during previous years include spectrohotometric,
chemiluminescence, electrochemical detection, chromatographic, capillary electrophoretic,
spectrofluorimetric methods. Out of these, methods based on spectroscopic detection of nitrate
have been discussed in this review article due to their easy availability, high sensitivity, low
detection limit, economical and facile nature. Methods based on spectrophotometry, Raman
Spectroscopy, IR and FTIR Spectroscopy, atomic absorption spectroscopy (AAS), fluorescence
spectroscopy, chemiluminescence, mass spectroscopy, molecular emission cavity analysis
(MECA), electron paramagnetic resonance spectrometry (EPR) and nuclear magnetic resonance
spectrocopy (NMR) have been reviewed. The basic principle, detection limits, detection range,
RSD%, sample throughput/h, advantages and disadvantages have been discussed.
Keywords: nitrate, spectrophotometric detection, spectroscopy, chemiluminescence and review

1. Introduction

Nitrite and nitrate are of paramount significance in the environmental samples like water, soil,
food and agriculture products. These ions are frequently analyzed to find out the level of
pollution. These chemical species have been used as a fertilizer component, food additives, etc.
Modernization of agriculture has resulted in excessive use of inorganic fertilisers which leads to
deterioration of the ecosystem. Nitrate can be readily reduced to nitrite in-vivo which interferes
with body oxygen transport mechanism by the irreversible conversion of haemoglobin to
methemoglobin. This causes great reduction in the oxygen exchange mechanism of haemoglobin
[1, 2]. Accelerated eutrophication takes place in fresh water and marine ecosystems due to
continuous addition of nitrates in the aquatic systems as a result of anthropogenic activities. [3,
4]. Higher concentrations of nitrate lead to methemoglobinemia in aquatic animals [5] and blue
baby syndrome in human infants [6]. Nitrite produces carcinogenic N-nitrosamines by reacting
with secondary or tertiary amines present in the human body [7, 8]. Nitrite plays a significant
role in diverse aquaculture diseases and is harmful to humans as well, if present in drinking
water. Nitrite ion is an important intermediate of the nitrogen cycle and occurs in soils as well as
surface water. Nitrite salts have numerous applications as corrosion inhibitor, in dye
manufacturing and food industry.

The use of nitrate compounds has been restricted in food processing industries in many countries
due to these toxic effects. Therefore, the determination of nitrite is imperative for environmental
protection and public health. The presence of interfering radicals restricts the detection of nitrate
especially at low concentration. Various techniques like potentiometry, spectrophotometry,
spectrofluorometry, chemiluminescence, fluorimetry, polarography, coulometry, etc. have been
proposed for nitrate determination each having its own merits and demerits. Spectroscopic
methods have excellent detection limits and have facile protocols, hence are the most extensively
used ones for nitrate/nitrite determination.
Moorcroft et al. (2001) reported the strategies employed to facilitate the detection, determination
and monitoring of nitrate and/or nitrite in his review along with the advantages/disadvantages
and limitations of those techniques [9]. The available methods for nitrate determination are
broadly classified into three types on the basis of analytical approach as proposed by Sah (1994)
[10]. The first analytical approach is based on determining the nitrate content directly through
colorimetry, potentiometry, UV radiation absorption, chromatography, etc. The principle of
second approach is reduction of nitrate to ammonium or nitric oxide followed by detection of
product concentration through colorimetry, potentiometry, fluorimetry, coulometry, etc. The
third approach is based on determining the concentration change of a chemical species that reacts
with nitrate to form a complex.

Wang et al. (2017) have also reviewed various techniques developed during the past 15 years for
the determination of nitrite and/or nitrate along with the detection principles and various
parameters [11]. They have discussed the advantages and disadvantages and tabulated the
detection limits and range of those methods. The review concludes that spectrofluorimetric
methods have become more attractive due to its easy availability, high sensitivity and selectivity,
low limits of detection and low-cost.

2. Spectroscopic Techniques Employed for Nitrate and Nitrite Determination

2.1 UV/Visible Spectroscopic methods

UV spectroscopy is widely used method for determination of nitrate in water samples due to its
versatility, simplicity and feasibility. This method of analysis is based on the measurement of
absorbance due to nitrate at 210 nm. Chlorine, nitrite, iron (III) and organic matter produce
interferences in nitrate concentration determination due to absorption in the same region. The
American Public Health Association (1992) protocol measured difference in sample absorbances
at 220 and 275 nm using distilled water as a blank [12]. Since dissolved organic matter can also
absorb UV radiation at 220 nm and nitrate does not absorb at 275 nm. Therefore, former methods
employed for nitrate determination through direct UV spectroscopy dealt with absorbance
measurement at 210 nm and another wavelength in the region where nitrate does not absorb UV
radiation [13, 15]. The most suitable of these methods was proposed by Norman and Stucki
(1981) which was based on the principle of UV absorbance measurement before and after the
destruction of nitrate [16]. Cox (1980) has also classified the spectrophotometric methods of
nitrate determination, based on nitration of a phenolic compound, oxidation of an organic
compound by nitrate or reduction of nitrate to nitrite and determination via the sulfanilamide
method [17]. Whereas, Wang et al. (2017) have classified spectrophotometric method for
detection of nitrite and nitrate into Griess Assay, nitrosation-based and catalytic
spectrophotometric methods [11].

2.1.1 Catalytic-spectrophotometric methods

Various catalytic-spectrophotometric methods based on the catalytic effect of nitrite on oxidation

of some indicative species or organic dyes with suitable oxidizing agents, have been widely used
for the determination of nitrate and nitrite in food, water and biological samples [18-25].
Substituted phenothiazine derivatives, triarylmethane alkaline dyes, triarylmethane acidic dyes,
azo dyes, quinone imine dyes, etc. have been used as indicative species. Whereas, potassium
bromate, potassium chlorate, potassium permanganate, hydrogen peroxide, etc., have been used
as oxidizing agents. These methods are time taking and are susceptible to interferences caused by
Fe2+, Fe3+, Ag+, SO32−, Br− and I− ions [9] and depend much on reaction conditions such as
acidity, temperature and reagent dosage. Application of flow-injection analysis (FIA) to
catalytic-spectrophotometric methods improves their efficiency. FIA has been used for
simultaneous determination of nitrite and nitrate by Ensafi and Kazemzadeh (1999) [26]. Pettas
et al. (1998) applied the reaction rate method for nitrite determination by applying a stopped-
flow technique [27]. Similar studies were performed by Kazemzadeh and Ensafi (2001) and
Mousavi and Shirkhanloo (2009) [22, 28]. They also studied the effects of other cations and
anions on the nitrite determination. Table 1 presents the comparison of various catalytic-
spectrophotometric methods available for nitrite and nitrate detection.

2.1.2 Nitrosation-based spectrophotometric methods

Several indicating species react with nitrite to give coloured products in case of nitrosation
reaction for determination of nitrite. Aydın et al. (2005) developed a spectrophotometric method
for the determination of nitrite in water based on its reaction with barbituric acid in acidic
solution to produce violuric acid, a nitroso derivative [29]. Beer’s law is obeyed at 310 nm, over
the nitrite concentration range of 0.00-3.22 ppm. Burakham et al. (2004) also employed
nitrosation reaction between nitrite ion and phloroglucinol (1,3,5-trihydroxybenzene) and
spectrophotometric detection at 312 nm, for nitrite as well as nitrate quantification in water
samples [30]. The system used a flow-injection analysis having a throughput of 20 samples/hr
with a relative standard deviation of less than 1.5%. The procedure is susceptible to interference
by Fe3+.

2.1.3 Griess Assay-based Methods

The Griess reaction was first reported by Johann Peter Griess in 1879 for nitrate determination in
saliva through diazotisation reaction. The Griess reaction has been used to detect bacterial
infection in the urogenital tract for a considerable period of time. The major nitrogen oxide anion
found in human urine is nitrate which is reduced to nitrite by bacterial reduction. This nitrite is
identified through Griess reaction. This reaction has been continuously modified and automated
in order to improve the assay and to combat interferences. The Griess assay is based on
diazotisation of an appropriate aromatic amine by acidified nitrite followed by coupling reaction.
A highly coloured azo chromophore is thus produced whose concentration is determined
spectroscopically, in the range 500–600 nm, in order to assess the nitrite concentration.

Various modifications have been made to this basic procedure, such as assay conditions, reagents
detection procedure, etc. Sulphanilic acid, nitroaniline and p-aminoacetophenone were the
commonly used target amine whereas phenol, 1-naphthol, 1-naphthol-4-sulphonate, 1-amino
naphthalene and 1,3-diaminobenzene were employed as potential coupling agents. The Griess
reaction is specifically used for nitrite determination. But nitrate can also be determined by this
method through chemical or enzymatic reduction of nitrate to nitrite before diazotization
reaction. This method of nitrate determination is easy, inexpensive and feasible. The detection
limits for Greiss assay ranges between 0.02 - 2 M, depending on the specific reagents chosen.

Fig. 1 : Schematic representation of the basic principle of Griess assay.

Wang et al. (1998a) determined nitrate and nitrite in water and some fruit samples using column
preconcentration based on the diazotization-coupling reaction between sulfanilamide or
sulfamethizole or sulfadimidine and sodium 1-naphthol-4-sulfonate in a hydrochloric acid
medium followed by reduction of nitrate to nitrite using the Cd–Cu reductor column [31].
Miranda et al. (2001) determined nitrate and nitrite concentrations simultaneously through
reduction of nitrate by vanadium (III) combined with detection by the acidic Griess reaction [32].
The sensitivity of the method was reported to decrease in presence of excess nitrite and hence
advocates its removal before nitrate assay. Tsikas (2007) analyzed nitrite and nitrate in biological
fluids through assays based on the Griess reaction and post-column derivatization of
chromatographically separated nitrite and nitrate through HPLC [33]. Badiadka and Kenchaiah
(2009) determined nitrite and nitrate in water, soil and pharmaceutical samples by the reaction of
sulfanilic acid with methyl anthranilate. Nitrate reduction was done in the presence of Zn/NaCl
which was subsequently diazotized with sulfanilic acid followed by coupling with methyl
anthranilate to give an azo dye [34]. Tu et al. (2010) published a simple miniaturised
photometrical method for rapid determination of nitrate and nitrite in freshwater samples [35].
All procedures for water analysis were performed in a 96-well microplate which allowed at least
48 samples to be simultaneously analysed in duplicate. Principle involved the reduction of nitrate
ion by copperised cadmium granules. Robledo et al. (2014) initially determined nitrite
concentration by using Griess reaction. Then, vanadium (III) was used for reduction of the nitrate
ion to nitrite (> 95%) which was also detected by the Griess reagents, that had been added
already [36]. They also compared the obtained results with the established Cd column method.

Gaurav et al. (2015) reported the effect of non-ionic surfactant, TritonX 100 in the range 1 ppm-
20 ppm as nitrate detection is interefered due to the presence of dissolved organic matter, nitrite,
hexavalent chromium and surfactant [37]. They did not consider the effect of cationic and
anionic surfactant because of their null effect on nitrate determination in water. Abbas and
Mostafa (2000) used solid phase spectrophotometry based on rapid sorption of dye formed from
nitrite through the Griess reaction into a thin layer of polyurethane foam (PUF) [38]. Nitrate was
pre-reduced using a cadmium reductor before applying the Griess reaction. Miro et al. (2000)
reported sequential injection spectrophotometric analysis of nitrite in natural waters using an on-
line solid-phase extraction and preconcentration method [39]. Gapper et al. (2004) used an
isocratic, ion-exchange chromatographic method with spectrophotometric detection for
determination of nitrite and nitrate concentration in dairy products [40]. This technique has
enhanced sensitivity and was free from endogenous UV-absorbing matrix.

Bastian et al. (1957) and MacDonald and Haddad (1970) have reported nitrate determination in
five per cent perchloric acid and hydroxide solutions, respectively, based on the ultraviolet
absorption [41, 42]. But the result obtained from these methods is highly affected by high
chloride concentration. Keeney et al. (1970) reported the use of a nitrate-selective ion electrode
method which appears to be the simplest but high concentrations of chloride affects the electrode
response thus reducing the sensitivity [43]. Lambert and DuBois (1971) proposed a
spectrophotometric method for determination of nitrate in the presence of chloride in water
samples [44]. Daniel et al. (1995) and Guerrero et al. (1996) used flow-injection analysis to
improve the precision and sensitivity for in situ nitrate and nitrite determination in sea water and
waste water or coastal marine water samples, respectively [45, 46]. Takeda and Fujiwara (1993)
reported the photo-induced conversion of nitrate without a copperized cadmium column with a
detection limit of 0.05 μM for nitrate [47].

Stanley et al. (1994) compared the analytical capabilities of a flow-injection analysis system with
an amperometric and a fibre optical sensor for nitrate determination as nitrite after reduction in a
cadmium column in river and well water with a detection limit of 50 μgL NO3- [48]. Ahmed et al.
(1996) also used flow-injection analysis for simultaneous determination of nitrite and nitrate
[49], whereas Schnetger and Lehners (2014) and Wang et al. (2016) used vanadium (III) chloride
as reductant instead of the well-known but toxic cadmium column, for environmental monitoring
and assessment [50, 51]. determined ammonia nitrogen in water by flow injection analysis based
on NH3-o-phthalaldehyde-Na2SO3 reaction [52]. Table 1 presents the list of reagents used in
some of the methods based on Griess assay for nitrate and nitrite determination along with the
comparison of various parameters.
 Table 1 : Comparison of various catalytic-spectrophotometric and Griess assay-based methods available for nitrite and/or nitrate detection.
S. Method Indicator Sample λmax Nitrite Nitrate FIA sample Reference
No. (nm) throughput/
Detection Detection RSD Detection Detection RSD h
limit range % limit range %
(μg/mL) (μg/mL) (μg/mL) (μg/mL)
1 Catalytic- Pyrogallol red Water/ 465 1.0 x 10-3 3.0 x 10-3- 1.60 1.0 x 10-2 3.0 x 10-2- 1.64 20 ± 3 Kazemzadeh and Ensafi
spectrophotometric Food 2.00 2.00 (2001) [22]
2 Catalytic- Chlorpromazine Water 525 1.2 0-1.0 x 10-1 1.8 N/A N/A N/A N/A Tomiyasu et al. (2001)
spectrophotometric [19]
3 Catalytic- Pyrogallol Water/ 465 1.6 x 10-3 2.4 x 10-3 2.8 0.003 4.0 x 10-3-1.0 3.7 10 Ensafi et al. (2004) [21]
spectrophotometric sulfonephthalein Food -1.6 x 10-1 x 10-1
4 Catalytic- Crystal violet Water 610 3.0 x 10-4 1.0 x 10-3 1.6 0.001 4.0 x 10-2 1.7 32 Yue et al. (2004) [20]
spectrophotometric - 5.0 x 10-2 - 2.0
5 Catalytic- Perphenazine Water 525 7.0 x 10-5 0 - 4.5 x 10- 1.1 N/A N/A N/A N/A Mubarak et al. (2007)
3
spectrophotometric [18]
6 Catalytic- Sulfonazo III Water/ 570 6.0 x 10-3 8.0 x 10-3 1.25 N/A N/A N/A 60 Nouroozi and Mirshafian
spectrophotometric Food - 0.3 (2009) [25]
7 Catalytic- Nuclear fast red N/A 518 7.0 x 10-1 2.0 1.75 N/A N/A N/A N/A Mousavi and Shirkhanloo
spectrophotometric - 4.5 x 101 (2009) [28]
8 Catalytic- Tropaeolin 00 Water 530 2 x 10-3 6.0 x 10-3 NA N/A N/A N/A N/A Moldovan (2010) [23]
spectrophotometric - 5.0 x 10-1
9 Griess assay Sulfanilamide + N-(l- Water 520 N/A N/A N/A N/A 5 x 10-3 –2.5 2.2 x 27 Henriksen and Selmer-
naphthyl)-ethylenediamine x 10-1 10-3 Olsen (1970) [54]
10 Griess assay Sulfanilic acid + 1- Water 520 N/A N/A N/A 2.0 x 10-2 0.0-5 N/A 30 Al-Wehaid and
Naphthylamine Townshend (1976) [56]
11 Griess assay 4-Aminobenzotri fluoride + N/A 510 N/A 0.8-20 N/A N/A N/A N/A N/A Amin (1986) [53]
1-Naphthol
12 Griess assay Sulfanilamide + N-(l- soil + plant 540 N/A N/A N/A N/A N/A N/A N/A Singh (1988) [55]
naphthyl)-ethylenediamine extracts
13 Griess assay Sulfanilamide + 1- Water + Food 543 1.4 x 10-6 2 x 10-6 0.48 1.1 x 10-6 1.5 x 10-6 N/A N/A Wang et al. (1998b) [58]
naphthol-4-sulfonate - 40 x 10-6 - 30 x 10-6
14 Griess assay Sulfamethizole + 1- Water + Food 532 1.2 x 10-6 2 x 10-6 0.56 0.9 x 10-6 1.5 x 10-6 N/A N/A Wang et al. (1998b) [58]
naphthol-4-sulfonate - 40 x 10-6 - 30 x 10-6
15 Griess assay sulfanilic acid + methyl Water + soil + 493 9.3 x 10-4 2 x 10-4 N/A N/A 2.6 x 10-4 N/A N/A Badiadka and Kenchaiah
anthranilate pharmaceutical - 8 x 10-3 - 1.07 x 10-2 (2009) [34]
preparations
2.1.4 Enzymatic-spectrophotometric Method

Borcherding et al. (2000) reported a microtiter plate-based nitrate detection in environmental

and medical analysis through enzymatic reduction of nitrate by dissimilatory nitrate
reductase from Pseudomonas stutzeri [59]. Sohail and Adeloju (2016) reviewed the available
biosensors for nitrate determination using the enzyme nitrate reductase (NaR) as excellent
non-toxic alternative to all other available analytical methods [60]. Vakilian and Massah
(2018) developed a portable nitrate biosensing device using electrochemistry and
spectroscopy [61]. The device contained plant-based nitrate reductase as biocatalyst,
anthraquinone-2-sulfonate as mediator and support vector machine as machine learning
method. It was usable upto 10 days after the enzyme immobilization and was able to predict
nitrate concentration in more than 200 samples without any enzyme replacement. They used
a machine learning method to learn the features of train samples in order to predict nitrate
concentration in liquid samples, such as drinking water, juice, plant extract and human saliva,
between 3.5 and 8.5 pH.

2.1.5 Chromatographic-spectrophotometric Methods

Reversed-phase high-performance liquid chromatography with UV spectrophotometric

detection of nitrate was used by Schroeder (1987) through an octadecyl column and aqueous
phosphoric acid/dihydrogen phosphate mobile phase with a detection limit of 0.007 mg/L
[62]. Hexavalent chromium and sulfate acted as slight positive interferences, negligible at
typical environmental concentrations. Lookabaugh and Krull (1988) also used similar
technique along with on-line post-column photolysis, ultraviolet absorbance and
electrochemical detection [63]. Flow-injection analysis was used by Devi et al. (1989), with
an on-line anion-exchange column and obtained a detection limit of 50 nM [64]. This method
was based on the principle of displacement of thiocyanate from a column and followed by its
reaction with Fe (III) to form a red complex having absorbance maxima at 480 nm. Everett et
al. (1995) analysed nitric oxide-derived nitrite and nitrate ions in biological fluids and
compared this method with conductimetric, spectrophotometric as well as capillary
electrophoretic techniques. HPLC with UV and electrochemical detection was also used by
Rizzo et al. (1998) to detect nitrite/nitrate balance during photoinduced cerebral ischemia in
rat [66]. Tsikas et al. (1999) analysed nitrite and nitrate in ultrafiltrate samples of human
plasma as S-nitroso-N-acetylcysteine with a detection limit of 50 nM nitrite [67]. They had
validated this method by gas chromatography-mass spectrometry. Papadoyannis et al. (1999)
reported a rapid, accurate, and sensitive method for simultaneous determination of nitrite and
nitrate in drinking water and human serum with detection limits of 0.1 ng and 0.2 ng for
nitrite and nitrate, respectively, per 20 μL injection [68].

Green et al. (1982) analyzed nitrate, nitrite and [15N] nitrate via reduction with a high-
pressure cadmium column in the samples of urine, saliva, deproteinized plasma, gastric juice
and milk through gas chromatography/mass spectrometry with a lower detection limit of 1.0
nmol NO3− or NO2−/mL [69]. This system allowed quantitative reduction of nitrate and
automatic elimination of interference due to the common compounds present in urine and
other biological fluids. Pratt et al. (1995) used multichannel flow injection analysis for nitrate
and nitrite in biological samples with a detection limit of 25 nM for both and compared the
results with that of the HPLC system [70]. Habu et al. (1994) and Yokoi et al. (1996)
analyzed nitrite, nitrate, and nitric oxide synthase activity in brain tissue by automated flow
injection technique [71, 72]. Ridnour et al. (2000) reported a spectrophotometric method for
direct detection and quantitation of nitric oxide, nitrite, and nitrate in cell culture media [73].

2.1.6 Reduction Method

Hilton and Rigg (1983) determined nitrate in lake water by adapting the hydrazine-copper
reduction method for use on a discrete analyser [74]. They also studied the performance
statistics and instrument-induced difference from segmented flow conditions. Zatar et al.
(1999) used phosphomolybdenum blue complex for determination of nitrite and nitrate [75].
This method was based on reduction of phosphomolybdic acid to phosphomolybdenum blue
complex by sodium sulfide which is further oxidized by nitrite causing decrease in intensity
of the blue color at 814 nm. The proposed method is applicable for nitrite and nitrate
concentration determination in water, meat products and vegetables. Several other workers
have also reported spectrophotometric methods based on different principles for assessment
of nitrite and nirate [58, 76-88].
2.2 Raman Spectroscopic Methods

Raman spectroscopy is based on the principle of measuring the inelastic scattering of light
caused by molecular vibrations involving changes in polarization of the molecule. Kneipp et
al. (2002) reported the utility of this technique in finding the molecular structure and
intermolecular interactions of a compound alongwith the fingerprint molecular signals [89].
Raman spectrometry has been employed for water analysis by several researchers [90-97].

Haverbeke and Brown (1978) reported that water is a very weak Raman scatterer [98].
Kowalchyk et al. (1995) used normal Raman spectroscopy as an on-line detector for capillary
electrophoresis to detect sub-ppm mixtures of nitrate and perchlorate in water [99].
Kauffmann et al. (2015) proposed a method for quantitative detection of inorganic salt,
sodium nitrate in aqueous solutions [100]. They used OH stretching band as internal standard
for nitrate determination. Li et al. (2016) presented various methods for nitrate quantification
in his review based on applications of Raman spectroscopy in detection of water quality
[101]. Comparison of various Raman-spectroscopic methods available for nitrite and nitrate
detection has been done in Table 2.

2.2.1 Laser Raman Spectroscopy-based Methods

Cunningham et al. (1977) studied the influence of experimental parameters on detection

sensitivity for laser Raman analysis of dissolved solutes nitrate, sulfate, carbonate,
bicarbonate, monohydrogen phosphate, dihydrogen phosphate, acetate ion and acetic acid in
water [102]. They derived an equation expressing the signal-to-noise ratio in terms of solute
concentration, measurement time, spectral slit width, laser power fluctuations and
background intensity of solvent. Haverbeke and Brown (1978); Thibeau et al. (1978) used
laser excited Raman spectroscopy for detection of water pollutants [98, 103]. The detection
limits ranged between 25-75 mg/L for strongly scattering anions such as NO3-, SO42-, etc.
When they changed the scattering geometry from the normal 90º to 180º, simulating a remote
sensing instrument, their sensitivity for NO3- changed from 25 to 150 mg/L.
2.2.2 Resonance Raman Spectroscopy-based Methods

Furuya et al. (1980) determined nitrite ion in waste and treated waters by resonance Raman
spectrometry for optimum control of waste water treatment [104]. It was based on the
transformation of NO2− into a colored azo dye which was analysed through resonance Raman
spectrometry. Addition of sodium thiocyanate as quencher reduces the background intensities
markedly. Lombardi et al. (1994) employed Raman spectroscopy using a CCD detector for
detection of nitrates, nitrites and sulfates, both in the solid phase as well as aqueous solution
[105]. The position and width of major nitrate peak at 1050 cm-1 (approx.) shifted with
concentration and this feature could be used for quantitative detection of nitrates. Lanoul et
al. (2002) used 204 and 229 nm excited UV Resonance Raman spectroscopic detection as an
excellent tool for on-line monitoring of nitrate and nitrite in wastewater treatment processes
on a real time basis [106]. The presence of other chemical species does not interfere with the
measurements of NO2-/NO3- Raman bands.

2.2.3 Surface-enhanced Raman spectroscopy (SERS)-based Methods

Surface-enhanced Raman spectroscopy (SERS) is rapid, sensitive, less destructive and

noninvasive technique useful for in situ and in vitro analysis of chemicals [107]. Nie and
Emory (1997); Kneipp et al. (1997) and Moskovits (2005) focused on the ability of SERS to
detect samples on nanosubstrates with detection limits as low as parts per billion level or
even single molecule [108-110]. Gajaraj et al. (2013) employed surface-enhanced Raman
spectroscopy (SERS) method coupled with a commercially available gold coated silicon
material nanosubstrate for detection of nitrate and nitrite in water and wastewater due to the
inert nature of gold [111]. Raman signals were enhanced by a factor of about 104 by the
application of this method as compared to the Raman spectroscopy. Phosphate appeared to be
the major interfering anion for nitrate measurement.

Mosier-Boss and Lieberman (2000 and 2003) reported the detection of nitrate and sulfate
anions in aqueous environments by Normal Raman Spectroscopy and Surface-Enhanced
Raman Spectroscopy (SERS) detection of cationic-coated, silver substrates [112, 113]. The
adsorption of anions on cationic-coated SERS substrates is depicted through Frumkin
isotherm. Appreciable detection limits were obtained for the sensors by using lasers
operating in the visible light range at a high power output along with long acquisition times,
anion preconcentration or use of multi-pass cells. Such sensors have limitations such as their
size, lack of durability, longer analysis times, and/or power requirements.

Durickovic and Marchetti (2014) detected nitrate as polyvalent alternative for water pollution
detection [114]. Luo et al. (2014) applied the rGO/TAgA/Rh6G sensor to probe the changes
in Rh6G concentration based on the nitrosation reaction between nitrite and the Rh6G
molecular probe [115]. Chen et al. (2016) developed a highly sensitive and selective method
to detect the nitrite concentration in pond water, a synthetic urine solution and pickle brine,
using Fe3O4@SiO2/Au magnetic nanoparticles (MNPs) by surface-enhanced Raman
scattering (SERS) [116]. 4-aminothiophenol (4-ATP) molecules conjugated on
Fe3O4@SiO2/Au MNPs were triggered by nitrite ions in acidic media, to form azo bonds.

2.2.4 Shell-Isolated Nanoparticle-Enhanced Raman Spectroscopy (SHINERS)-based

Methods

Shell-isolated nanoparticle-enhanced Raman spectroscopy (SHINERS) was developed by Li

et al. (2010 and 2011) [117, 118]. It consisted of a gold nanoparticle with an insulating thin
SiO2 shell (Au/SiO2) as SERS active substrates. SHINERS act as the next generation of
advanced spectroscopy [119], applicable for environmental monitoring, life science, food
safety, etc. But, it cannot be used for detection of atomic cations and anions directly or other
organic molecular species with an extremely low SHINERS cross-section. Zhang et al.
(2013) also reported the determination of trace NO2- in tap and lake water samples using
SHINERS [120]. The substrate contained Au/SiO2 nanoparticles with pinholes. The nitrite
concentration could be detected indirectly on the basis of diazotization-coupling reaction.
The detection limits were 0.07, 0.08 and 0.10 ppm at 1137, 1395 and 1432 cm-1, respectively.
This technique shows great potential as an analytical tool for trace analysis of nitrite in food,
biological and environmental samples.

2.2.5 Morphology-Dependent Stimulated Raman Scattering (MDSRS)-based Methods

MDSRS is a nonlinear spectroscopy which utilizes the set of macroscopic natural

electromagnetic oscillation modes, commonly known as morphology-dependent resonances
(MDRs), of micron-sized axisymmetric particles in order to enhance Raman scattering from
molecules present in the particle. The spatial and spectral positions of MDRs are determined
by particle size, shape and refractive index [121, 122]. Aker et al. (1999) detected nitrate ion
in aerosols using morphology-dependent stimulated Raman scattering (MDSRS) [121]. They
determined nitrate ion concentration of 5x10-5 M in 180 mm diameter aqueous aerosols. The
experimental intensities were well comparable with signal intensities predicted using the
MDSRS gain equation.
 Table 2 : Comparison of various Raman-spectroscopic methods available for nitrite and/or nitrate detection.
S. Principle Sample Nitrate/ Raman Detection limit Detection range RSD Reference
No. Nitrite emission (μg/mL) (μg/mL) %
(cm-1)
1 Laser Raman Spectrometry Water Nitrate N/A N/A N/A N/A Cunningham et al.
(1977) [102]
2 Laser Raman Spectrometry Water Nitrate 1045 2 x 103 6 x 103-105 < 0.06 Furuya et al. (1979)
ppm [123]
3 Rapid Normal Raman Spectroscopy Water Nitrate 1047 N/A 6.2 x 102 N/A Kowalchyk et al.
(1995) [99]
4 Morphology-Dependent Stimulated Raman aerosols Nitrate 1047 3.1 N/A N/A Aker et al. (1999)
Scattering (MDSRS) [111]
5 Cationic thiols coated, silver SERS Water Nitrate 1047 2.6 x 105 104-107 N/A Mosier-Boss and
substrates Lieberman (2000)
cysteamine hydrochloride (CY), [112]
dimethylaminoethanethiolhydrochloride
(DMA), diethylaminoethanethiol
hydrochloride (DEA), L-cysteine ethyl ester
hydrochloride (CYE), 2-mercapto-4-
methylpyrimidine hydrochloride (MMP)
6 UV Resonance Raman Spectroscopy Water Both NO3- - 1044 < 2 x 102 1 x 102-5 x 104 N/A Lanoul et al. (2002)
NO2- - 1325 [106]
7 Shell-Isolated Nanoparticle- Enhanced Food, Nitrite N/A 1 x 10-2 5 x 102-6 x 103 14.5% Zhang et al. (2013)
Raman Spectroscopy (SHINERS) biological and [120]
environmental
samples
8 Surface-Enhanced Raman Spectroscopy Water Nitrate 1056 5 x 10-1 1-104 N/A Gajaraj et al. (2013)
(SERS) [111]
9 Surface-Enhanced Raman Spectroscopy Water Nitrite 1505 9.2 x 10-6 3.22 x 10-5 - 3.31 4.4- Luo et al. (2014)
(SERS) based on nitrosation reaction of x 10-3 7.5 [115]
rhodamine 6G molecular probe
2.3 IR Spectroscopic Methods

Infrared Spectroscopy involves the study of interaction of infrared light with molecules, as
they absorb frequencies, characteristic of their structure. IR absorptions take place at resonant
frequencies that match the vibrational frequency and are affected by shape of the molecular
potential energy surfaces, atomic masses and the associated vibronic coupling. This
technique is mainly used to determine functional groups present in a molecule and has been
used on a large scale for quantification of nitrite and nitrate by various researchers which are
being discussed hereafter.

Magee et al. (1964) and Yang and Low (1973) used infrared spectroscopy and infrared
internal reflectance spectrometry, respectively for quantitative analysis of aqueous
nitrite/nitrate solutions [125]. The former method could detect nitrate even in the presence of
other anions, without prior separation. Falk (1990) determined nitrate, nitrite and ammonium
ions even at nanomole concentrations based on ion exchange and matrix isolation in alkali
metal halide pressed pellets [126]. He also suggested the applicability of the proposed
method to other polyatomic ions. Jiao and Lips (2000) determined nitrate concentration along
with estimation of kinetic parameters of net nitrate uptake by barley (Hordeum vulgare)
seedlings through multiple wavelength spectroscopy [127].

Linker and Shaviv (2006) used transmittance mid-infrared spectroscopy based on adsorption
of nitrate onto anion exchange membrane immersed in the sample solution, followed by
determination of amount of nitrate sorbed on the membrane [128]. Sulfate affects the
spectrum of charged membrane but does not interfere with nitrate determination. Goebbert et
al. (2009) presented an infrared photodissociation spectra of the microhydrated nitrate ions
NO3-(H2O)1-6, between 600 to 1800 cm-1 [129]. Huang et al. (2017) determined total
nitrogen, ammonia and nitrite in river water by near-infrared spectroscopy and chemometrics
[130].

2.4 FTIR Spectroscopic Methods

Fourier-transform is a mathematical process, required to convert raw data into actual

spectrum. The technique based on such transformation is called Fourier-Transform Infrared
spectroscopy. It is used to obtain infrared spectrum of absorption or emission of a solid,
liquid or gas. It has significant advantage over a dispersive spectrometer, viz., higher signal-
to-noise ratio and less sensitivity to stray light and a wide spectral range. In aqueous solution,
various inorganic materials can be brought in a form suitable for infra-red spectral analysis
[131]. Aqueous solution spectra have several advantages over solid state spectra in case of
most of the common polyatomic inorganic ions.

Goulden and Manning (1967) obtained calibration curves for nitrate and phosphate in water
and pointed out the possibility of study of pH-induced structural changes and the formation
of complex ions in aqueous solution [131]. Alijah and Mooney (1969) demonstrated the use
of Attenuated Total Reflection (ATR) method for quantitative detection of polyatomic anions
in aqueous solution and also compared the corresponding frequencies in the solid state and
Raman frequencies [132]. Elzinga et al. (2001) used ATR-FTIR spectroscopy for direct
investigation of the absorption of polyanions on mineral thin surfaces in aqueous systems
placed in contact with the ATR crystals [133]. Strauss et al. (2002) improved the detection
limits of anionic compounds in water significantly by coating common ATR devices with
highly selective ion-exchange compounds [134].

The development of silver halide fibers has promoted the implementation of Fiberoptic
Evanescent Wave Spectroscopy (FEWS) for direct monitoring of nitrate in environmental
systems as these fibres, transmit into the mid-IR with minimal loss [135]. Spectra of
phosphate, carbonate, sulfate, ammonium and soil organic constituents were studied to find
out any possible interference offered to nitrate determination. Direct transmission of MIR
radiation through partially loaded ion-exchange membranes was also found effective. The
fiber could be used for about 10 determinations only due to its low mechanical strength and
deterioration.

Linker et al. (2004) reported Fourier Transform Infrared–Attenuated Total Reflection nitrate
determination of soil pastes [136]. The spectral region around the nitrate band (1300–1550
cm-1) was analyzed by principal component regression (PCR), partial least squares (PLS) and
cross-correlation with reference libraries inclusive of the spectra of pure ions and/or soils.
Jahn et al. (2006) also used FTIR-ATR with mid-infrared (mid-IR) to detect nitrate in various
soil types both in the laboratory and field [137]. Choe et al. (2010) used derivative analysis
and sample treatments for determination of soil nitrate based on similar method [138]. The
key advantages of this method were simple sample preparation, short measurement time and
a wide range of applications. Shao et al. (2014 and 2017) analyzed different nitrogen isotope
labelled nitrates using mid-infrared attenuated total reflectance spectroscopy [139, 140].
Various IR and FTIR spectroscopic methods have been compared in Table 3.
 Table 3 : Comparison of various IR and FTIR spectroscopic methods available for nitrate and/or nitrite detection.
S. Principle/Reagent used IR/ Sample Nitrate/ IR Wave number Detection limit Detection RSD Reference
No FTIR Nitrite (μg/mL) range %
. (μg/mL)
1 Nitron (1,4-diphenyl-3,5- IR Nitrate 730 and 748 nm 103 N/A 5% Magee et al. (1964) [124]
enedianflohydroxytriazole)
2 Infrared Internal Reflectance IR Water Nitrate + NO3- - 1352 cm-1 NO3- - 6.2 x 101 N/A N/A Yang and Low (1973)
Spectrometry Nitrite NO2- - 9.2 x 102 [125]
3 Matrix isolation in pressed alkali IR Water Nitrate + Nitrate-1384.27 cm-1 1.86 x 10-4 4.34 x 10-4 2% Falk (1990) [126]
metal halide pellets Nitrite Nitrite - 1271.5 cm-1 - 3.96 x 10-2
4 Fiberoptic Evanescent Wave FTIR Soil + Water Nitrate 1350 and 1250 cm-1 104 0 - 3 x 105 N/A Shaviv et al. (2003)
Spectroscopy (FEWS) [135]
5 Principal Component Regression, FTIR Soil Nitrate 1370 cm-1 N/A 0-1.094 x 106 N/A Linker et al. (2004) [136]
Partial Least Squares, and Cross-
Correlation Analysis
6 Fourier Transform Infra Red (FTIR) FTIR Soil Nitrate 1370-1380 cm-1 N/A 0-106 N/A Jahn et al. (2006) [137]
Attenuated Total Reflectance (ATR)
technique
7 Anion exchange membrane IR Soil + Water Nitrate 1040 cm-1 < 1.55 N/A N/A Linker and Shaviv
and transmittance mid-infrared (2006) [128]
spectroscopy
8 Comparison with calculated IR Microhydrated Nitrate 600 to 1800 cm-1 N/A N/A N/A Goebbert et al. (2009)
B3LYP/augcc-pVDZ harmonic Nitrate Ions [129]
frequencies NO3-(H2O)1-6
9 Derivative Analysis and Sample FTIR Soil Nitrate 1400-1200 cm−1 N/A N/A N/A Choe et al. (2010) [138]
Treatments
10 Fourier Transform Infrared FTIR Soil + Water Nitrate 1200-1500 cm–1 N/A N/A N/A Shao et al. (2014) [139]
Attenuated Total Reflection
Spectroscopy
Principal Component
Regression (PCR) and Partial Least
Squares Regression (PLSR)
11 Near-Infrared Spectroscopy and IR Water Nitrite 4000-12500 cm-1 N/A 5 x 10-2 N/A Huang et al. (2017) [130]
Chemometrics, Principal component - 3.14 x 101
analysis
2.5 Atomic Absorption Spectroscopic Methods

Atomic absorption spectrometry (AAS) is a spectroanalytical technique that measures the

concentration of elements upto parts per billion of a gram (µg dm–3) in a sample at a
wavelength specifically absorbed by that element. It can be used for estimation of over 70
different elements in solution, or directly in solid samples and is used in pharmacology,
biophysics, toxicology research, etc. Kumamaru et al. (1965) used atomic absorption
spectrophotometry for the determination of nitrate ions based on complexation of the ions
with copper neocuproine followed by extraction with methyl isobutyl ketone (MIBK) and
measured the concentration of complexed copper [141]. The extracted species had the
chemical formula [Cu(Neocup.*)2NO3]. This method was further modified for indirect
determination of nitrate, nitrite and nitro groups by Houser and Fauth (1970) [142]. Nitro and
nitrite groups were oxidized to nitrate by cerium (IV) sulfate or potassium permanganate and
nitrate was determined by following the proposed method. Nitro and nitrite groups could be
reduced to elemental nitrogen by sulfamic acid, thus permitting nitrate determination in the
presence of other two types of compound.

Yamamoto et al. (1969) also used solvent extraction of the ion-pair of bis-
(neocuproine)copper(I) nitrate [143]. The copper concentration was determined by atomic
absorptiometry in an air-acetylene flame at the 3247Å copper line. They also studied the
effect of diverse anions on nitrate extraction and the composition of the extracted species
along with the extraction percentage. Atomic absorption spectroscopy coupled with flow
injection analysis (FIA-AAS) was used by Gallego et al. (1986) [144]. Interferences were
observed due to thiocyanate, perchlorate and chlorate ions. Silva et al. (1986) employed
liquid-liquid extraction and a flow injection system based on the reaction of NO3- with
bis(2,9-dimethyl-l,10-phenanthrolinato)-copper(I) [145]. This method was found unsuitable
for soil and plant extracts due to interference by various ions, especially chloride. Atomic-
absorption spectrometric, potentiometric and polarographic methods were used by Hassan
(1981) for the determination of organic and inorganic nitrates in the presence of various other
ions [146]. This method was based on reduction of nitrate with cadmium metal and 0.1 M
hydrochloric acid through a four-electron reduction process, thus converting it into nitrous
oxide. They did not observe any interference even by 100-fold mass excess of C1-, C2O42,
CO32-, and NH4+ anions and cations.
Alonso et al. (1998) used an on-line cadmium microcolumn for nitrate reduction to nitrite
thus oxidizing metallic cadmium to Cd (II), which is measured by electrothermal atomic
absorption spectrometry (ETAAS) [147]. The sample was passed through an Amberlite
cation-exchange resin in order to reduce the interferences, offered by commonly occurring
ions in water samples, such as Na+, Ca2+ and Mg2+ to a minimum level. The authors used
direction opposite to that of sample loading, for washing the column. Anwar and Haque
(1985) reported a method based on formation of insoluble ternany [Ag(Phen)2]NO3 (1 : 2 : 1)
complex by adding an excess of [Ag(Phen)2] solution in sample containing nitrate [148]
followed by absorbance measurement for unreacted silver ions in air-acetylene flame. This
method was effectively applicable for nitrate determination in fertilizer samples. Anwar et al.
(1986) used mercury-sulphuric acid mixture for the reduction of nitrate followed by atomic
absorption spectrophotometric determination of mercurous ion thus released in air-acetylene
flame [149]. Some of these methods have been compared in Table 4.
 Table 4 : Comparison of various atomic absorption spectroscopic methods available for nitrate and/or nitrite detection.
S.No. Reagent used Sample λmax (nm) Nitrate/ Detection limit Detection range RSD% Reference
Nitrite (μg/mL) (μg/mL)
1 Copper neocuproine N/A 456 Nitrate N/A N/A N/A Kumamaru et al. (1965) [141]
2 Bis-(neocuproine)copper(I) N/A N/A Nitrate N/A N/A N/A Yamamoto et al. (1969) [143]
3 Cerium (IV) sulfate or Inorganic and N/A Both 1 N/A N/A Houser and Fauth (1970) [142]
potassium permanganate organic nitrates
4 Cadmium metal and 0.1 M Inorganic and 228.8 Nitrate N/A 3.1 x 10-2 - 3.1 1 Hassan (1981) [146]
hydrochloric acid organic nitrates
5 [Ag(Phen)2] solution Fertilizer 328.1 Nitrate N/A 3.1 x 10-2 - 3.1 x 10-1 0.45 Anwar and Haque (1985) [148]
6 Mercury-sulphuric acid mixture N/A N/A Nitrate N/A N/A N/A Anwar et al. (1986) [149]
7 Bis(2,9-dimethyl-l,10- Food 328.1 Both 4.0 x 10-2 for 1.3 x 10-1 - 2.20 NO3- - 3.7 Silva et al. (1986) [145]
phenanthrolinato)-copper(I) both NO3- and NO2- - 4.1
NO2-
8 On-line cadmium microcolumn Water 228.8 Nitrate 5.0 x 10-2 5.0 x 10-1 - 1.4 x 101 2.3 Alonso et al. (1998) [147]
2.6 Fluorescence Spectroscopic methods

Fluorescence spectroscopy or spectrofluorometry analyzes fluorescence from a sample using

a beam of light, usually ultraviolet light for exciting the electrons in molecules of certain
compounds leading them to emit light which is typically, but not necessarily, visible light. It
is commonly used in various fields for analyzing organic compounds. Most of the
flurospectrometry based methods for nitrate and nitrite quantification are based on reducing
nitrate to nitrite with a suitable reductant usually copperized reductant, Cu-Cd reductant
column or enzyme systems. Then, the nitrite was determined on the basis of its chemical
reactivity (nitrosation/diazotization) with different non-fluorescent compounds, like indole,
safranin O, acetaminophen, 2,3-diaminonaphthalene, N-methyl-4-hydrazino-7-
nitrobenzofurazan, 5,6-diamino-1,3-naphthalene disulfonic acid (DANDS), etc. to give
fluorescent chemical species, which were easily detected with fluorescence spectroscopy.
Fluorimetric methods are very sensitive having detection limit as low as 1 x 10-8 M NO3-
[150].

The fluorimetric methods proposed by Dombrowsky and Pratt (1972), Hornyak and
Szekelyhidi (1983) and Ohta et al. (1986) involved several steps in preparation of fluorescing
products, rendering these methods unsuitable for routine analysis [151-153]. Lee and Field
(1984) performed postcolumn fluorescence detection of nitrite, nitrate, thiosulfate and iodide
anions in high-performance liquid chromatography based on their reaction with Ce (IV) to
produce the fluorescent Ce (III) [154]. This method is simple but is susceptible to the
presence of other redox species. Motomizu (1987), Gladilovich et al. (1989) and Fernández-
Argüelles et al. (2004) have also used fluorescent spectroscopy for nitrate and/or nitrite
detection [150, 155, 156].

Lumpp et al. (1992) have developed an optical sensor having a poly(vinylchloride) (PVC)
membrane impregnated with a nitrate-selective ionophore as well as a proton-selective
(chromo)-ionophore, for the continuous monitoring of nitrate in aquatic environments [157].
Nitrate concentration is determined spectroscopically by protonation of indicator anion (Ind).
The advantages of the proposed sensor technique over potentiometric are minimal electrical
interference, elimination of baseline variation by using multi-wavelength calibration and
reference material is also not needed. Liu et al. (2013) used fluorescent gold nanoclusters for
selective and ultrasensitive detection of nitrite. Fluorescence spectroscopic methods can be
classified on the basis of mechanism, nitrosation/diazotization involved in detection of nitrate
and nitrite [158].

2.6.1 Nitrosation-Based Fluorescence Spectroscopic Methods

Nakamura (1980) used resorcinol as fluorimetric reagent for the determination of nitrate
whereas Ohta et al. (1986) determined nitrite in saliva by nitrosation of 4-hydroxycoumarin
in acidic medium followed by reduction to fluorescent nitroso derivative, 3-amino-4-
hydroxy-coumarin [159, 153]. The fluorescence intensity may be enhanced or quenched due
to the nitrosation reaction depending linearly on the nitrite concentration. Biswas et al.
(2004) analysed the ultra trace amount of nitrite and nitrate in air, water, soil and forensic
samples based on the ability of nitrite to quench the native fluorescence of murexide
(ammonium purpruate) [160]. Chloride ion interferes with nitrite determination and hence
needs to be removed by precipitating with silver sulphate. Biswas et al. (2004); Liu et al.
(2009), Zhang et al. (2003); Gao et al. (2005) and Wang et al. (2012) also used nitrosation-
based spectrofluorimetric methods for nitrate and/or nitrite determination [160-164].

2.6.2 Diazotization-Based Fluorescence Spectroscopic Methods

The fluorimetric methods developed by Axelrod and Engel (1975) and Gladilovich et al.
(1989) were significant, as fluorescing products were obtained in one step from 5-
aminofluorescein and 2,3- diaminonaphthalene, respectively [165, 155]. Lapat et al. (1997)
have reported the use of two fluorometric methods for nitrite detection based on reaction of
nitrite with either 2-amino-4-chloro-1-hydroxybenzene-6-sulphonic acid or 4-
aminofluorescein to form a diazonium salt which can be manipulated to produce highly
fluorescent complexes [166]. These methods have been successfully applied for detection of
the explosive 1,3,5-trinitro-1,3,5-triazacyclohexane (RDX) after its decomposition to nitrite.

Buldt and Karst (1999) used microplate fluorescence spectroscopy and HPLC for nitrite
determination. Jie et al. (1999) developed a fluorescence quenching method for nitrite
determination in water and food samples based on its reaction with indole to produce a
compound having no fluorescence in acidic medium [168]. Huang et al. (2000) presented the
kinetic spectrofluorimetric determination of nitrite in water samples and nitrogen dioxide in
the atmospheric samples through liquid droplet method [169]. Wang et al. (2000); Helaleh
and Korenaga (2000); Geetha and Balasubramanian (2000) and Guo et al. (2013) also used
spectrofluorimetric method for nitrite determination [170-173]. Li et al. (2000) used
reversed-phase high-performance liquid chromatography (RP-HPLC) along with
fluorescence [174].

Masserini Jr. and Fanning (2000) devised a fluorescence based sensor package for
simultaneous determination of nanomolar concentrations of nitrite, nitrate and ammonia in
seawater [175]. The background fluorescence from leachates and naturally occurring
dissolved organic matter, were corrected through Reverse Flow Injection Analysis (rFIA).
Fujii et al. (2004) used a microfluidic device for determination of sulfite and nitrite in
aqueous samples [176]. Martínez-Tomé et al. (2009) immobilized 2, 3-
diaminonaphthalene/cyclodextrin Complexes in a Sol–Gel Matrix as a new fluorimetric
sensor for nitrite [177]. Huang et al. (2008) determined ultra-trace level of nitrite in human
saliva [178]. Lin et al. (2017) determined nitrate in seawater with valve-free continuous flow
analysis method and established its corresponding instrument with only two multi-channel
pump for delivering the sample and reagent [179]. The proposed method and instrument were
less costly and easy to operate as compared with flow injection analysis. Some of the
spectrofluorimetric methods have been reported in Table 5 [180-185].
 Table 5 : Comparison of various fluorescence-spectroscopic methods available for nitrite and/or nitrate detection.
S. Reagents Used Mechanism Sample Nitrate/ Wavelength Detection Detection RSD% Ref.
No. Nitrite limit range
(μmol/L) (μmol/L)
λex λem
(nm) (nm)
1 Ce( IV) Reduction Water Both N/A 214.9 NO2- - 1.39 x 1-100 <5 Lee and Field (1984)
10-1 [154]
NO3- - 1.46 x
10-1
2 3-amino- N/A Water Nitrate N/A N/A 1 10-300 2 x 10-5 M Motomizu et al.
1,5naphthalenedisulphonic acid (1987) [150]
3 2,3-diaminonaphthalene (DAN) Intra-molecular Water/Cell culture Both 375 415 1.0 x 10-2 1.25 x 10-2 NO2- - 0.42 Li et al. (2000) [173]
diazotization media/Plasma/Urine – 2.0 NO3- - 0.56
4 5,6-diamino-1,3-naphthalene Intra-molecular Water Nitrite 297.6 428.8 2.0 x 10-3 2.0 x 10-2 1.7 Wang et al. (2000)
disulfonic acid (DANDS) diazotization - 2.4 [170]
5 Tetra-substituted amino Diazotization Water Nitrite 610 678 7 2.1 x 101-8.4 3.2 Zhan et al. (2001)
aluminum phthalocyanine x 102 [180]
(TAAlPc)
6 Neutral red Diazotization Food Nitrite 548 609 2.0 x 10-1 9.0 x 10-1-4.3 1.7 Li et al. (2003) [181]
7 Rhodamine 110 Nitrosation Water Nitrite 502.4 521.6 7.0 x 10-4 1.0 x 10-2-3.0 2.14 Zhang et al. (2003)
x 10-1 [162]
8 Murexide (ammonium Nitrosation Water/Soil/Forensic Both 349.0 444.5 1.0 x 10-2 1.0 x 10-1- 0.4878 Biswas et al. (2004)
purpurate) samples 2.17 x 101 [160]
9 Proflavin N/A Water Nitrite 290 519 2.0 x 10-2 0-8.7 0.3 Fernandez-Arguelles
et al. (2004) [156]
10 2,3-diaminonaphthalene (DAN) Intra-molecular Water Nitrite 360 407 4.0 x 10-1 0-3.0 x 101 1.3 Fujii et al. (2004)
diazotization [176]
11 1,3,5,7-tetramethyl-8-(4'- Diazotization Water/Food Nitrite 497 510 6.5 x 10-4 8.0 x 10-3-3.0 4.53 Li et al. (2004) [183]
aminophenyl)-4,4-difluoro-4- x 10-1
bora-3a,4a-diaza-s-indacence
(TMABODIPY)
12 Mono[6-N(2-carboxy-phenyl)]- Nitrosation Water/Soil/ Nitrite 330 440 0.0002 2.0 x 10-2-1.7 1.38 Gao et al. (2005)
β-cyclodextrin (OACCD) Food [163]
13 1,3,5,7-tetramethyl-2,6- Intra-molecular Food Nitrite 500 510 0.00021 9.0 x 10-3-3.0 2.4 Huang et al. (2006)
dicarbethoxy-8-(3',4'- diazotization x 10-1 [184]
diaminophenyl)-
difluoroboradiaza-s-indacence
(TMDCDABODIPY)
14 2-Amino-5, 7-dimethyl-1, 8- Diazotization Water Nitrite 346 403 0.0406 1.0 x 10-1-2.5 0.23 Chen et al. (2007)
naphthyridine (ADMND) [182]
15 1, 3, 5, 7-tetramethyl-8-(3, 4- Diazotization Saliva Nitrite 498 507 0.0001 6.0 x 10-3-5 x 1.12 Huang et al. (2008)
diaminophenyl)- 10-1 [178]
difluoroboradiaza-s-indacene
(DAMBO)
16 Unsymmetrical Nitrosation Water Nitrite 538 561 0.0002 1.0 x 10-2-3.5 2.36 Liu et al. (2009) [161]
rhodamine x 10-1
17 2,3-diaminonaphthalene (2,3- Intra-molecular N/A Nitrite 382 408 0.02 0-10 N/A Martinez-Tome et al.
DAN)/Hydroxypropyl-β- diazotization and (2009) [177]
cyclodextrin (HP-β-CD) 430
18 Rh 6G-SiO2 Nitrosation N/A Nitrite 240- 418 1.2 2-60 N/A Wang et al. (2012)
400 [164]
19 o-phenylenediamine (OPD)/ Intra-molecular Food/Soil Nitrite 420 568 0.3 9.0 x 10-1- 1.38 Guo et al. (2013)
Hydroxypropyl-β-cyclodextrin diazotization 1.74 x 101 [174]
(HP-β-CD)
20 Gold nanoclusters (AuCNs) N/A Water Nitrite 350 670 0.001 2.0 x 10-2-5.0 < 3.5 Liu et al. (2013) [158]
x 101
21 Folic acid Diazotization Water Nitrite 365 370 0.018 5.0 x 10-2-3.2 N/A Lu et al. (2015) [185]
x 101
2.7 Chemiluminescence-based Methods

Chemiluminescence or chemoluminescence is the emission of light or limited emission of

heat, as a result of chemical reactions. The major difference between chemiluminescence and
fluorescence or phosphorescence is that the electronic excited state is the product of a
chemically exothermic reaction rather than that of photon absorption. It is used for the
analysis of inorganic and organic species, biomolecular assay, DNA sequencing, etc. It has
also been utilized for determining small amounts of impurities or poisons present in air, for
example, NO determination with detection limits down to 1 ppb [17, 186].

2.7.1 Gas-phase chemiluminescence-based Methods

Nitrite and nitrate determination through this technique are based on their reduction to nitric
oxide which is further determined from its chemiluminescent reaction with ozone. The
equations for respective half reactions are:

The standard reduction potentials for NO3- and NO2- are similar. Still NO2- can be reduced
under much milder conditions as compared to NO3-. Gaseous NO subsequently reacts with
ozone to produce nitrogen dioxide in an excited state (NO2*) and molecular oxygen.

The excited NO2* then decays giving a weak IR chemiluminescence above 600 nm. This
method can be used for the analysis of nitrite as well as nitrate. The nitrate concentration can
be determined from the difference but stronger reductant is required.
Fig. 2 : Schematic representation of basic chemistry of Gas-Phase Chemiluminescence
technique.

Cox (1980) reported that nitrite is selectively reduced under mild conditions while stronger
reduction conditions are required for the total nitrate-nitrite content determination, except
when vanadium (II) was used as a reducing agent [17]. Interference is not produced by
suspended material and color hence their removal before analysis is not essential. He used
acetic acid–sodium iodide (NaI) mixture as reductant for acid decomposition of NO2- to nitric
oxide. Ammonium molybdate [Mo (VI)] and ferrous ammonium sulfate [Fe (II)] in 50%
concentrated sulfuric acid (hot) was used for reduction of both NO2- and NO3- to NO. NO3-
concentration was then determined from the difference of results obtained by the two
methods. This method was applied to analyses of seawater by Garside [187, 188]. Later
Braman and Hendrix (1989) indicated that V (III) reduces NO3- to NO and not V (II) [189].
Nitric oxide was removed from the solution by scrubbing with helium carrier gas and
followed by detection with chemliumlnescence NO analyzer. This method was advantageous
with nanogram detection limit because highly acidic solutions were not required for nitrate
reduction and is applicable to variety of environmental and biological samples like water,
sediment, plant materials, human urine and blood serum.

Walters et al. (1978) used acid dismutation reaction with acetic acid to produce nitric oxide
[190]. Doerr et al. (1981) used sodium ascorbate and tartaric acid in order to reduce NO2- to
nitric oxide in meat [191]. Nitrostable compounds, such as primary and secondary amines
and some reductants may interfere with the yield of nitric oxide. Yoshizumi and Aoki (1985)
used 40-95 wt % phosphoric acid (H3PO4) or 55-68 wt % sulfuric acid (H2SO4) in flow
injection determination for reduction of NO3- and NO2- to nitric oxide [186]. Yang et al.
(1997) examined the efficacy of five reducing agents [V (III), Mo (VI) + Fe (II), NaI, Ti (III),
and Cr (III)] to reduce nitrite and/or nitrate to nitric oxide along with the effect of
temperature [192]. Lingane and Pecsok (1949) and Cresser (1977) used chromium (II)
[Cr(II)] and titanium (III) [Ti(III)] in acidic medium for nitrate reduction [193, 194]. Garside
(1982) and Cox and Frank (1982) applied this technique to the analysis of nitrite in seawater
and blood/urine, respectively [187, 195].

Chemiluminescence has been used along with FIA assemblies by several researchers [196-
198, 192]. Sen et al. (1994) reduced nitrite with potassium iodide, in foods and biological
materials by flow injection or high-performance liquid chromatography with
chemiluminescence detector [199]. They reported that chemiluminescence methods are much
superior to the colorimetric one due to its speed, versatility, sensitivity and minimum sample
preparation. Aoki et al. (1997) used aqueous sample itself as the carrier stream in which
reagents were injected in a flow injection analysis method [200]. Iodide and titanium (III) in
acidic media were used to reduce nitrite and nitrate, respectively into nitric oxide.

In his previous work Aoki (1990) stripped nitric oxide, reduced by iodine in acidic media,
from aqueous phase through a gas-permeable membrane and the gas-phase
chemiluminescence (CL) was detected after the reaction of NO with ozone [200]. The
microporous poly(tetrafluoroethylene) tube was used as separator to transfer nitric oxide into
gas phase. Aoki and Wakabayashi (1995) and Dunham et al. (1995) also reported a similar
method for simultaneous determination of nitrite and nitrate [201, 202]. Since the detection
occurs in gas-phase, only volatile conversion products swept to the gas phase. Therefore,
sample coloration and turbidity did not interfere. Nagababu et al. (2007) determined plasma
nitrite without interferences of other nitric oxide-related chemical species such as S-
nitrosothiols [196].

2.7.2 Luminol chemiluminescence-based Methods

Reagents like lucigenin, tris(2,2-bipyridyl)ruthenium (II), luminal (5-amino-2,3-

dihydrophthalazine-1,4-dione) and peroxyoxalate derivatives based chemiluminescence
methods are also used for determination of nitrate and nitrite. The most common among them
is the oxidation of luminol in alkaline medium. 3-aminophthalate anion is produced which
gives strong chemiluminescence at 425 nm [203]. Oxidants such as permanganate,
hexacyanoferrate (III) or hydrogen peroxide are generally used for the nitrite determination.
Icardo et al. (2001) determined nitrite indirectly with a flow-injection assembly, based on the
inhibition of oxidation of acriflavine [204]. The acriflavine solution and nitrite mixture was
merged with the oxidant solution, potassium permanganate in sulfuric acid medium. The
resulting chemiluminiscence was inhibited by nitrite after reaction with aminoacridine. This
method was applied to residual waters, industrial formulations and soil samples for nitrite
determination.

Mikuska and Vecera (2003) used a fully automated flow-injection system for the
simultaneous determination of nitrite and nitrate in water [205]. The method is based on
online photolytic conversion of nitrate to nitrite and its detection after oxidation to
peroxynitrous acid by the chemiluminescent reaction with luminol. The cationic interferences
were eliminated by passing the sample through cation-exchange column whereas common
anions did not interfere. Yue et al. (2006) determined femtogram nitrite in human urine
samples using a sensitive flow injection luminol–myoglobin chemiluminescence system
[206]. Nitrite bound myoglobin produced the ferric heme nitrite complex, which further
catalyzed the electron transfer from luminol to myoglobin, thus producing fast
chemiluminescence. The chemiluminescence increase was proportional to the nitrite
concentration.

He et al. (2007) applied microflow injection analysis (μFIA) system on a chip for the nitrite
determination in some vegetables and fruits [207]. The reaction of ferrocyanide and nitrite in
acidic medium produces ferricyanide which further reacts with luminol to produce
chemiluminescence. Yaqoob et al. (2012) followed luminol chemiluminescence based
detection of nitrate and nitrite [208]. Nitrate reduction was done through copperized
cadmium (Cu–Cd) column. The nitrite thus produced reacted with acidic hydrogen peroxide
to form peroxynitrous acid. The oxidation of luminol in an alkaline medium in the presence
of the peroxynitrite anion produced chemiluminescence. The effect of cations such as Ca (II),
Mg (II), Co (II), Fe (II) and Cu (II) was masked online with EDTA. Common anions like
PO43-, SO42- and HCO3- did not interfere. The effect of salinity was also investigated. Table 6
presents the comparison of various chemiluminescence methods available for nitrate and/or
nitrite detection.
 Table 6 : Comparison of various Chemiluminescence methods available for nitrate and/or nitrite detection.
S. No. CL system Reagents Used Nitrate/ Sample Absorbance Detection limit Detection range RSD % FIA sample Reference
Nitrite maxima, (μmol/L) (μmol/L) throughput/
λmax (nm) h
1 NO/O3 NaI and acetic acid Both N/A N/A NO2- - 1.08 x 10-3 NO2- - 0.1086-5.43 NO2- - 2 12 Cox (1980) [17]
4% ferrous ammonium sulfate NO3- - 8.06 x 10-4 NO3- - 0.0016-1.209 NO3- - 3
and 2% ammonium molybdate
-2 -2 2
2 NO/O3 4% ferrous ammonium sulfate Nitrate N/A N/A 1.61 x 10 1.61 x 10 -1.61 x 10 N/A 20 Yoshizumi et al. (1985)
and 2% ammonium molybdate [186]
-
3 NO/O3 Vanadium (III) Both Water/sediment/ N/A NO2- - 4.347 x 10 N/A N/A N/A Braman and Hendrix
plant materials/ 2 -2 (1989) [189]
-6.521 x 10
human urine/ -2
NO3- - 3.22 x 10 -
Blood serum 4.84 x 10
-2
4 NO/O3 aqueous iodide in acid Nitrite Water 540 8.7 x 10-4 N/A 2 60-180 Dunham et al. (1995)
[202]
2 3
5 NO/O3 Vanadium (III) [V (III)], Both Blood plasma 600 N/A 4.0 x 10 – 2.0 x 10 N/A NO2- - 30 Yang et al. (1997) [192]
Molybdenum (VI) [Mo (VI)] + NO3- - 15
Iron (II) [Fe (II)],
Sodium iodide (NaI),
Titanium (III) [Ti(III)], and
Chromium (III) [Cr (III)]
-2
6 NO/O3 Iodide and Both Water N/A 1.0 x 10 N/A NO2- - 0.6 20 Aokia et al. (1997) [197]
Titanium (III) in aqueous NO3- - 6.7
media
-1 3
7 NO/O3 Vanadium (III) + detection by Both Biological 540 N/A 5.0 x 10 -1.0 x 10 N/A N/A Miranda et al. (2001)
the acidic Griess reaction samples (NO3-) [32]
-2 -2 1
8 NO/O3 Hydrazine and/or ascorbic acid Both Water N/A 1.0 x 10 5.0 x 10 -5.0 x 10 3.2 26 Kanda and Taira (2003)
[198]
-2 -2
9 NO/O3 glacial acetic acid and ascorbic Nitrite Plasma N/A 1.0 x 10 1.0 x 10 -5 8.1 N/A Nagababu and Rifkind
acid (2007) [196]
-1 -1 1
10 Luminol KMnO4 and Nitrite Water/industrial N/A 2.2 x 10 2.2 x 10 -1.7 x 10 1.4 76 Icardo et al. (2001) [204]
acriflavin formulations/
soil
-3 -3 1
11 Luminol H2O2 Both Water 210 2.0 x 10 8.0 x 10 -1.0 x 10 2.1 24 Mikuska and Vecera
(2003) [205]
-1 4
12 Luminol myoglobin Nitrite Human urine 409 4.0 x 10 1.0-2.2 x 10 2.6 120 Yue et al. (2006) [206]
-2 -1
13 Luminol K3Fe(CN)6 Nitrite Food 520 9.0 x 10 1.7 x 10 -2.2 4.1 30 He et al. (2007) [208]
14 Luminol H2O2 Both Water 425 NO2- -2.1 x 10-4 NO2- -2.1 x 10-4 – 1.086 1.8-4.6 NO2- - 90 Yaqoob et al. (2012)
NO3- - 3.2 x10-4 NO3- - 3.2 x 10-4 – 8.06 NO3- - 40 [208]
x 10-1
2.8 Mass Spectrocopic Methods

Mass spectroscopy is an analytical technique that ionizes chemical species by bombarding it

with electrons and sorts the ions based on their mass-to-charge ratio. The plot of ion signal
vs. mass-to-charge ratio gives the mass spectrum. It is used in various fields and can be
applied to pure samples as well as complex mixtures in order to elucidate their structure and
chemical properties. Mitschke et al. (2007); Helmke and Duncan (2006) and Tsikas (2000);
Kage et al. (2000) have reported the use of gas chromatography-mass spectroscopy (GC-MS)
for nitrate and/or nitrite determination [209-212]. Tsikas (2000) performed simultaneous
quantification of nitrite and nitrate in biological fluids, plasma and urine of humans by
GC/MS [211]. It is based on single derivatization of endogenous nitrite and nitrate and the
15
externally added N-labeled analogues simultaneously in aqueous acetone by
pentafluorobenzyl bromide to the nitro and nitric acid ester pentafluorobenzyl derivatives,
respectively. This method enables discrimination between S-nitro- and S-nitroso-glutathione
and study of metabolism and reactions of NO and its metabolites in vitro and in vivo and is
interference-free.

Kage et al. (2002) determined nitrate and nitrite in human plasma through gas
chromatography-mass spectrometry, using extractive alkylation with pentafluorobenzyl
bromide, using tetradecyldimethylbenzyl ammonium chloride as the phase-transfer catalyst,
with 1,3,5-tribromobenzene as internal standard [213]. The lower limit of detection for both
compounds was 0.005 μmol/mL. They determined nitrate and nitrite concentrations in blood
of six victims who committed suicide by automobile exhaust gas inhalation. Akyuz and Ata
(2009) reported the application of GC–MS as well as liquid chromatography with
fluorescence detection (LC-FL) methods for determination of low level nitrite and nitrate in
biological, food and environmental samples [214]. Whereas, Campanella et al. (2012)
presented a novel isotope dilution gas chromatography method for the nitrate determination
in vegetables with a detection limit of 2 μg/g [215].

2.9 Molecular emission cavity analysis (MECA)-based Methods

Molecular emission cavity analysis (MECA) is based on the analysis of highly coloured
solutions through standard spectroscopic techniques. Nitrite is reduced to NO and nitrogen
gas carries it into the flame followed by monitoring of the emission at 640 nm. Spectral
interferences are offered by carbon dioxide and hydrogen sulphide, which are emitted within
the cavity. But this could be removed through precipitation as a suitable metallic sulphide.
Honma and Smith (1954) have determined nitrite and nitrate by measuring the cyanogen
emission at 3883 Å [216].

Belcher et al. (1981) determined ammonium- and nitrate-nitrogen by rapid, precise and
interference free molecular emission cavity analysis at parts per million to percentage levels
in fertiliser, river water and sewage samples by injecting the sample solution on to solid
sodium hydroxide in a small vial [217]. The generated ammonia was carried by nitrogen into
MECA oxy-cavity and the intensity of the NO-O band was measured at 500 nm. Al-Zamil
and Townshend (1982) applied MECA for nitrite and nitrate determination after their
reduction to nitrogen monoxide by iodide or zinc followed by measurement of white
emission stimulated from nitrogen monoxide at 526 nm [218]. Celik and Henden (1989) used
a cavity containing an entire flame. Nitrite was converted to nitrogen monoxide by iodide, for
its analysis [219]. Whereas, nitrate was reduced to nitrite through copperised cadmium
column and was determined as nitrite.

2.10 Photoacoustic spectroscopy (PAS)-based Methods

Photoacoustic spectroscopy (PAS) is based on detection of pressure waves caused by

absorption-induced sample heating. Linker (2008) determined nitrate concentration in soil
via photoacoustic spectroscopic analysis of ion exchange membranes [220]. As ion
exchangers, either membranes or resins mimic the physical processes of nutrient uptake by
plants, they can be used to determine the presence of nutrients and pollutants in soils [221,
222]. Commercially available anion exchange membrane, typically used in water treatment
applications was loaded with Cl- prior to use by immersing overnight in a 1 M NaCl solution.
Weak correlations between nutrient sorption by ion exchangers and chemically extractable
nutrients have been reported. In the proposed method, the author focuses on directly
determining the amount of nitrate sorbed onto such membranes through mid-infrared PAS.
This method is suitable for wider range of membranes, including the membranes commonly
used for water treatment which may be too thick for transmittance measurements. Linker and
Shaviv (2006) reported that the transmittance spectra of the loaded membranes which do not
absorb mid-infrared radiation strongly can be directly analysed through photoacoustic
spectroscopy.

2.10 Electron paramagnetic resonance spectrometry (EPR)-based Methods

Electron paramagnetic resonance (EPR) or electron spin resonance (ESR) spectroscopy is a

technique analogous to nuclear magnetic resonance (NMR) spectroscopy. But, the electron
spins of materials with unpaired electrons are excited rather than their nuclear spins. It is
used for studying metal complexes or organic radicals. Pawe et al. (1990) used electron
paramagnetic resonance spectroscopic investigation for studying the inhibition of
phosphorwlastic system of Clostridum sporogenes by nitrite [223]. Wennmalm et al. (1991)
reported nitrite detection in nanomolar range by measuring the paramagnetic properties of
NO bound to heme-containing proteins [224]. Yordanov et al. (2001) performed consecutive
estimation of nitrate and nitrite in vegetables and fruits based on the selective reactions of
nitrate and nitrite ions to yield a stoichiometric amount of the EPR-active
mononitrosyldiethyldithiocarbamate complex of iron in aerobic conditions [225]. The
detection limit depends on the background concentration of nitrogen monoxide and dioxide
in the air. Whereas, Goetz et al. (2010) and Schwab et al. (2010) determined the affinity of
nitrite for methemoglobin [226, 227] and Bawn and MacMillan (2018) have studied the
nitrite binding in myoglobin through this technique [228].

2.11 Nuclear Magnetic Resonance Spectroscopy (NMR)-based Methods

Nuclear magnetic resonance is a nuclei specific spectroscopy that uses a large

magnet/magnetic field to probe the intrinsic spin properties of atomic nuclei. It uses radio
frequency waves to promote transitions between nuclear energy levels. It has numerous
applications throughout the physical sciences and industry like elucidation of molecular
structure. Lu et al. (2004) distinguished the sources of groundwater nitrate by 1H NMR of
dissolved organic matter (DOM) as it carries characteristic marks in its molecular structures
which can be recognized by spectroscopic analysis [229]. They carried this study at Chino
Basin of the Santa Ana River Watershed in southern California and reported that disposal of
dairy wastes on croplands is the primary source of nitrate contamination in groundwater in
the study area.
3. Comparison of different spectroscopic methods for nitrite/nitrate determination

Nitrate analysis is done by preparation of calibration graph and absorbance measurement of

the samples and standards at both 220 and 275 nm. UV-visible spectrophotmetric techniques
for nitrite and nitrate quantification have been classified as catalytic, nitrosation, enzymatic,
chromatographic, reduction and Griess assay-based methods. Out of these catalytic-
spectrophotometric methods are popular due to their versatility and sensitivity. Application
of flow-injection analysis (FIA) improves its efficiency. Another popular technique is Griess
assay based spectrophotometry due to its low cost and easy feasibility with detection limits
between 0.02 - 2 M. But poor sensitivity, time-consuming nature and ionic interferences
restrict the use of these methods.

Each chemical substance has a characteristic Raman fingerprint and easy quantitative
measurements, feasibility of remote and flow-through detection systems are some of the
advantages of using Raman spectroscopy for analysis of water pollutants [98]. It offers high
signal to noise spectra due to the absence of fluorescence interference from other species
[106]. Sample solutions are generally diazotized followed by coupling with appropriate
organic chemicals to form azo dye solutions which show absorbance in the wavelength range
of 400–500 nm. The percentage error for nitrate measurement in wastewater by Surface-
enhanced Raman spectroscopy (SERS) was comparable to that by ion chromatography and it
could also be used for simultaneous detection of sulfate [111]. Shell-Isolated Nanoparticle-
Enhanced Raman Spectroscopy (SHINERS) can be employed for the analysis of compounds
with weak Raman response, as well as indirect detection of iodate ion, phenols and aromatic
amines [117, 118]. Whereas, Morphology-Dependent Stimulated Raman Scattering
(MDSRS) spectroscopies are capable of providing information about absolute concentration,
but it has not been used as field measurement tool primarily due to too low detection
sensitivity. Mosier-Boss and Lieberman (2000) pointed out that organic matter and humics
present in water samples fluoresce with the use of visible excitation which may be about 100
times stronger than Raman scattering and hence can interfere with the Raman signal. Near IR
excitation is generally preferred in order to avoid such fluorescence interference. Whereas,
mid-IR spectroscopy is basically not much effective for quantitative analysis of ions in
aqueous medium in order to avoid strong absorbance by water molecules. In case of soil
samples, pastes were prepared from to enhance adhesion between the ATR crystal and
sample in order to mimic real field conditions. The concentration of the nitrate to be
measured was increased by adding 0.01 M CaSO4 solution as an extractant in the soil paste
since it does not interfere in the characteristic spectra of the analyte [138].

The commonly used atomic absorption spectroscopic procedure is based on transformation of

nitrate into complex with a metal ion, such as, copper or silver ion followed by its extraction
into a suitable solvent and determination through AAS. It suffers from interference offered
by various ions present in the sample, especially chloride. The detection limit of AAS has
been reported as 4.0 x 10-2 μg/mL [145]. Low reagent and sample consumption, wide pH
range, good sensitivity and selectivity are some of the advantages of this technique.

Spectroflurometric methods for nitrate and nitrite determination are based on reducing nitrate
to nitrite with a suitable reductant and nitrosation or diazotization of suitable non-fluorescent
reagents based on their chemical reactivity to give fluorescent chemical species. These
methods are very sensitive with a detection limit of about 1 x 10-8 M [150].
Spectrofluorimetric methods are simple, convenient, highly sensitive, selective, cheap having
detection limit as low as 1 x 10-8 M NO3-. Several fluorescent probes have been developed
for nitrite analysis. Vicinal aryl diamines are the most common among them but they suffer
from poor stability. Determination of Ce(III) resulting from nitrite oxidation with Ce(IV) is
also a popularly used method but is susceptible to the influence of other redox species.
Whereas, chemiluminescence suffers from the issues of low stability and reproducibility.
Nitrite and nitrate are reduced to nitric oxide which is determined from its chemiluminescent
reaction with ozone with detection limits of 103 [192]. Gas-phase chemiluminescence is
based upon monitoring the reaction between gaseous NO and ozone, hence it proves better
for the elimination of matrix effects. But this technique is highly complex and high
combustion temperatures (600°C) are required. Use of chemiluminescence along with FIA
assemblies, generates notably enhanced detection limits over conventional UV/Vis systems
[196-198, 192].

Linker and Shaviv (2006) reported that the transmittance spectra of the membranes
containing a thick layer for mechanical support, such as membranes used for water treatment,
which do not absorb mid-infrared radiation strongly can be readily analyzed through
photoacoustic spectroscopy (PAS) [128]. PAS technique is cheap, fast and offers reagent-free
estimation of the amount of ions sorbed onto the membrane, but it is not much preferred for
quantitative analysis and its sensitivity is also low as compared to other techniques.
Molecular emission cavity analysis (MECA) is preferable for the analysis of highly coloured
solutions as they can be dealt with standard spectroscopic techniques. It involves the use of a
cavity containing an entire flame thus eliminating the need for a burner system and hence
consumes lesser gases. Nitrite is reduced to NO and swept into the flame of the cavity by
nitrogen carrier gas followed by emission recording. Celik and Henden (1989) obtained
comparable results for nitrate content in drinking water through MECA and a standard
spectrophotometric method [219]. Wennmalm et al. have reported nitrite detection using
electron paramagnetic resonance spectrometry (EPR) in organ perfusates and human plasma
based on measuring the paramagnetic properties of NO bound to heme-containing proteins in
nanomolar range [103]. Nitric oxide is readily oxidized to nitrogen dioxide under
physiological conditions which forms nitrite and nitrate due to its high reactivity.

Conclusions

This review focuses on the spectrophotometric methods available for the determination of
nitrite and/or nitrate along with their merits and demerits. It can be concluded that while
determining nitrate concentration, the selection of method should be based on the suitability
of the method towards interferences present and analyte concentration in the sample. The
spectroscopic method of nitrate and nitrite determination is successful if water sample
contains little or no organic matter. Interferences due to the presence of other ions may affect
the sensitivity of these methods. UV spectroscopy can be used as a detection device in
combination with other sophisticated techniques like chromatography due to its low cost,
easy feasibility and high sensitivity. Different methods based on Raman spectroscopy, IR and
FTIR spectroscopy, atomic absorption spectroscopy, fluorescence spectroscopy, molecular
emission cavity analysis (MECA), photoacoustic spectroscopy (PAS), electron paramagnetic
resonance spectrometry (EPR) and chemiluminescence have also been discussed.
References

1. T.Y. Chan, Food-borne nitrates and nitrites as a cause of methemoglobinemia.

Southeast Asian J. Trop. Med. Public Health 27 (1996) 189-192.

2. J.M. Fahey, Pretreatment effects on nitrite-induced methemoglobinemia: saline and

calcium channel antagonists, Pharmacol. Biochem. Behav. 37 (1990) 457-459.

3. P.M. Vitousek, J.D. Aber, R.W. Howarth, G.E. Likens, P.A. Matson, D.W. Schindler,
W.H. Schlesinger, G.D. Tilman, Human alteration of the global nitrogen cycle: sources
and consequences. J. Ecol. Appl. 7 (1997) 737–750.

4. J.A. Camargo, A. Alonso, A. Salamanca, Nitrate toxicity to aquatic animals: a review

with new data for freshwater invertebrates. J. Chemosphere 58 (2005) 1255–1267.

5. F.B. Jensen, Nitrite disrupts multiple physiological functions in aquatic animals. J.

Comp. Biochem. Phys. A. 135 (2003) 9–24.

6. F.R. Greer, M. Shannon, Infant methemoglobinemia: the role of dietary nitrate in food
and water, J. Pediatrics, 116 (2005) 784–786.

7. H. Suzuki, K. Iijima, A. Moriya, K. McElroy, G. Scobie, V. Fyfe, K. McColl,

Conditions for acid catalysed luminal nitrosation are maximal at the gastric cardia, Gut,
52 (2003) 1095-1101.

8. K.O. Honikel, The use and control of nitrate and nitrite for the processing of meat
products, Meat sci. 78: 68-76 (2008).

9. M.J. Moorcroft, J. Davis, R.G. Compton, Detection and determination of nitrate and
nitrite: a review, Talanta 54 (2001) 785–803.

10. R.N. Sah, Nitrate-nitrogen determination-a critical review, Commun. Soil Sci. Plant
Anal. 25 (1994) 2841-2869.
11. Q.H. Wang, L.J. Yu, Liu, Y.; Lin, L.; Lu, R.; Zhu, J.P.; He, L.; Lu, Z.L.; Methods for
the detection and determination of nitrite and nitrate: A review. Talanta 165 (2017) 709-
720.

12. American Public Health Association (APHA), American Water Works Association
(AWWA), and Water Environment Federation (WEF). 1992. Standard Methods for the
Examination of Water and Wastewater 18th Edition.

13. E. Goldman, R. Jacobs, Determination of Nitrates by Ultraviolet Absorption, Am.

Water Works Assoc. 53 (1961) 187-191.

14. F.A.J. Armstrong, Determination of Nitrate in Water Ultraviolet Spectrophotometry.

Anal. Chem. 35 (1963) 1292–1294.

15. R.J. Norman, J.C. Edberg, J.W. Stucki, Determination of Nitrate in Soil Extracts by
Dual-wavelength Ultraviolet Spectrophotometry, Soil Sci. Soc. Am. J. 49 (1985) 1182-
1185.

16. R.J Norman, J.W. Stucki, The determination of nitrate and nitrite in soil extracts by
ultraviolet spectrometry, Soil Sci. Soc. Am. J. 45 (1981) 347-353.

17. R.D. Cox, Determination of nitrate and nitrite at the parts per billion level by
chemiluminescence, Anal. Chem. 5 (1980) 332–335.

18. A.T. Mubarak, A.A. Mohamed, K.F. Fawy, A.S. Al-Shihry, A novel kinetic
determination of nitrite based on the perphenazine-bromate redox reaction, Microchim.
Acta 157 (2007) 99-105.

19. T. Tomiyasu, Y. Konagayoshi, K. Anazawa, H. Sakamoto, A kinetic method for the

determination of nitrite by its catalytic effect on the oxidation of chlorpromazine with
nitric acid, Anal. Sci. 17 (2001) 1437-1440.

20. X.F. Yue, Z.Q. Zhang, H.T. Yan, Flow injection catalytic spectrophotometric
simultaneous determination of nitrite and nitrate, Talanta 62 (2004) 97-101.
21. A.A. Ensafi, B. Rezaei, S. Nouroozi, Simultaneous spectrophotometric determination of
nitrite and nitrate by flow injection analysis, Anal. Sci. 20 (2004) 1749-1753.

22. A. Kazemzadeh, A.A. Ensafi, Simultaneous determination of nitrite and nitrate in

various samples using flow-injection spectrophotometric detection. Microchem. J. 69
(2001) 61-68.

23. Z. Moldovan, Kinetic Spectrophotometric Determination of Nitrite with Tropaeolin 00-

Bromate System, Anal. Lett. 43 (2010) 1344-1354.

24. J. Ghasemi, A. Jabbari, A. Amini, A. Oskoei, B. Abdolahi, Kinetic spectrophotometric

determination of nitrite based on its catalytic effect on the oxidation of methyl red by
bromate, Anal. Lett. 37 (2004) 2205-2214.

25. S. Nouroozi, R. Mirshafian, Flow injection kinetic spectrophotometric method for the
determination of trace amounts of nitrite, Talanta 79 (2009) 1149-1153.

26. A.A. Ensafi, A. Kazemzadeh, Simultaneous determination of nitrite and nitrate in

various samples using flow injection with spectrophotometric detection. Anal. Chim.
Acta 382 (1999) 15-21.

27. I.A. Pettas, S.I. Lafis, M.I. Karayannis, Reaction rate method for determination of nitrite
by applying a stopped-flow technique, Anal. Chim. Acta 376 (1998) 331-337.

28. H.Z. Mousavi, H. Shirkhanloo, Spectrophotometric determination of nitrite based on its

catalytic effect on the reaction of nuclear fast red and potassium bromated, J. Serb.
Chem. Soc. 74 (2009) 985–992.

29. A. Aydın, Ö. Ercan, S. Taşcıoğlu, A novel method for the spectrophotometric

determination of nitrite in water, Talanta 66 (2005) 1181-1186.

30. R. Burakham, M. Oshima, K. Grudpan, S. Motomizu, Simple flow-injection system for

the simultaneous determination of nitrite and nitrate in water samples, Talanta 64 (2004)
1259-1265.
31. G.F. Wang, M. Satake, K. Horita, Spectrophotometric determination of nitrate and
nitrite in water and some fruit samples using column preconcentration. Talanta 46
(1998) 671-678.

32. K.M. Miranda, M.G. Espey, D.A. Wink, A rapid, simple spectrophotometric method for
simultaneous detection of nitrate and nitrite, Nitric Oxide 5 (2001) 62–71.

33. D. Tsikas, Analysis of nitrite and nitrate in biological fluids by assays based on the
Griess reaction: Appraisal of the Griess reaction in the l-arginine/nitric oxide area of
research, J. Chromatogr. B. 851 (2007) 51–70

34. N. Badiadka, S. Kenchaiah, A Spectrophotometric Method for the Determination of

Nitrite and Nitrate, Eurasian J. Anal. Chem. 4 (2009) 204-214.

35. X. Tu, B. Xiao, J. Xiong, X. Chen, A simple miniaturised photometrical method for
rapid determination of nitrate and nitrite in freshwater. Talanta 82 (2010) 976-983.

36. E.G. Robledo, A. Corzo, S. Papaspyrou, A fast and direct spectrophotometric method
for the sequential determination of nitrate and nitrite at low concentrations in small
volumes, Mar. Chem. 162 (2014) 30-36.

37. K. Gaurav, P. Devi, B. Kumar, S. Bansod, Study of Effect of Interferent in the

Determination of Nitrate in Water, Aquat. Procedia 4 (2015) 1094-1098.

38. M.N. Abbas, G.A. Mostafa, Determination of traces of nitrite and nitrate in water by
solid phase spectrophotometry, Anal. Chim. Acta 410 (2000) 185-192.

39. M. Miro, A. Cladera, J.M. Estela, V. Cerda, Sequential injection spectrophotometric

analysis of nitrite in natural waters using an on-line solid-phase extraction and
preconcentration method, Analyst 125 (2000) 943-948.

40. L.W. Gapper, B.Y. Fong, D.E. Otter, H.E. Indyk, D.C. Woollard, Determination of
nitrite and nitrate in dairy products by ion exchange LC with spectrophotometric
detection, Int. Dairy J. 14 (2004) 881–887.
41. R. Bastian, R. Weberling, F. Palilla, Ultraviolet Spectrophotometric Determination of
Nitrate...Application to Analysis of Alkaline Carbonates, Anal. Chem. 9 (1957) 1795-
1797.

42. J.C. MacDonald, L. Haddad, Ultraviolet spectrophotometric determination of nitrite-

nitrate in potassium hydroxide, Environ. Sci. Technol. 4 (1970) 676-678.

43. K.E. Keeney, B.H. Byrnes, J.J. Genson, Determination of nitrate in waters with the
nitrate-selective ion electrode, Analyst, 95, 383-386 (1 970).

44. R.S. Lambert, R.J. DuBois, Spectrophotometric determination of nitrate in the presence
of chloride, Anal. Chem. 43 (1971) 955-957.

45. A. Daniel, D. Birot, M. Lahaitre, J. Poncin, Characterization and reduction of

interferences in flow-injection analysis for the in situ determination of nitrate and nitrite
in sea water, Anal. Chim. Acta 308 (1995) 413-424.

46. R.S. Guerrero, C.G. Benito, J.M. Calatayud, Flow-injection analysis-spectrophotometric

determination of nitrite and nitrate in water samples by reaction with proflavin, Talanta
43 (1996) 239-246.

47. K. Takeda, K. Fujiwara, Determination of nitrate in natural waters with the photo-
induced conversion of nitrate to nitrite, Anal. Chim. Acta 276 (1993) 25-32.

48. M.A. Stanley, J. Maxwell, M. Forrestal, A.P. Doherty, B.D. MacCraith, D. Diamond,
J.G. Vos, Comparison of the analytical capabilities of an amperometric and an optical
sensor for the determination of nitrate in river and well water, Anal. Chim. Acta 299
(1994) 81-90.

49. M.J. Ahmed, C.D. Stalikas, S.M. Tzouwara-Karayanni, M.I. Karayannis, Simultaneous
spectrophotometric determination of nitrite and nitrate by flow-injection analysis,
Talanta 43 (1996) 1009-1018.

50. B. Schnetger, C. Lehners, Determination of nitrate plus nitrite in small volume marine
water samples using vanadium(III)chloride as a reduction agent, Mar. Chem.160 (2014)
91-98.
51. S. Wang, K. Lin, N. Chen, D. Yuan, J. Ma, Automated determination of nitrate plus
nitrite in aqueous samples with flow injection analysis using vanadium (III) chloride as
reductant, Talanta 146 (2016) 744-748.

52. Y. Liang, C. Yan, Q. Guo, J. Xu, H. Hu, Spectrophotometric determination of ammonia

nitrogen in water by flow injection analysis based on NH3- o-phthalaldehyde -Na2SO3
reaction, Anal. Chem. Res. 10 (2016) 1-8.

53. D. Amin, Determination of nitrite ion using the reaction with 4-aminobenzotrifluoride
and 1-naphthol. Analyst 111 (1986) 1335-1337.

54. A. Henriksen, An automatic method for determining nitrate and nitrite in fresh and
saline waters, Analyst 90 (1965) 83-88.

55. J.P. Singh, A rapid method for determination of nitrate in soil and plant extracts. Plant
and Soil 110 (1988) 137-139.

56. A. Al-Wehaid, A. Townshend. Spectrometric flow-injection determination of nitrate

based on reduction with titanium (III) chloride. Anal. Chem. Acta. 186 (1986) 289-294.

57. E. Szekely, Spectrophotometric determination of nitrate with p-

diaminodiphenylsulphone and diphenylamine-p-diaminodiphenylsulphone, Talanta 14
(1967) 941-950.

58. G.F. Wang, K. Horita, M. Satake, Simultaneous Spectrophotometric Determination of

Nitrate and Nitrite in Water and Some Vegetable Samples by Column Preconcentration.
Microchem. J. 58 (1998) 162-174.

59. H. Borcherding, S. Leikefeld, C. Frey, S. Diekmann, P. Steinrucke, Enzymatic

Microtiter Plate-Based Nitrate Detection in Environmental and Medical Analysis, Anal.
Biochem. 282 (2000) 1-9.

60. M. Sohail, S.B. Adeloju, Nitrate biosensors and biological methods for nitrate
determination, Talanta 153 (2016) 83-98.
61. K.A. Vakilian, J. Massah, A Portable Nitrate Biosensing Device Using Electrochemistry
and Spectroscopy, IEEE Sensors Journal, 18 (2018) 3080-3089.

62. D.C. Schroeder, The Analysis of Nitrate in Environmental Samples by Reversed-Phase

HPLC, J. Chromatogr. Sci. 25 (1987) 405-408.

63. M. Lookabaugh, I.S. Krull, Determination of nitrite and nitrate by reversed-phase high-
performance liquid chromatography using on-line post-column photolysis with
ultraviolet absorbance and electrochemical detection, J. Chromatogr. 452 (1988) 295-
308.

64. S. Devi, A. Townshend, Determination of nitrate by flow-injection analysis with an on-

line anion-exchange column, Anal. Chim. Acta 225 (1989) 331-338.

65. S.A. Everett, M.F. Dennis, G.M. Tozer, V.E. Prise, P. Wardman, M.R.L. Stratford,
Nitric oxide in biological fluids: analysis of nitrite and nitrate by high-performance ion
chromatography, J. Chromatogr. A 706 (1995) 437-442.

66. V. Rizzo, L. Montalbetti, A.L. Rozza, W. Bolzani, C. Porta, G. Balduzzi, E. Scoglio, R.

Moratti, Nitrite/nitrate balance during photoinduced cerebral ischemia in the rat
determined by high-performance liquid chromatography with UV and electrochemical
detection, J. Chromatogr. A. 798 (1998) 103-108.

67. D. Tsikas, S. Rossa, J. Sandmann, J.C. Frolich, High-performance liquid

chromatographic analysis of nitrite and nitrate in human plasma as S-nitroso-N-
acetylcysteine with ultraviolet absorbance detection. J. Chromatogr. B. 724 (1999) 199-
201.

68. I.N. Papadoyannis, V.F. Samanidou, C.C. Nitsos, simultaneous determination of nitrite
and nitrate in drinking water and human serum by high performance anion-exchange
chromatography and UV detection, J. Liq. Chromatogr. R.T. 22 (1999) 2023-2041.

69. L.C. Green, D.A. Wagner, J. Glogowski, P.L. Skipper, J.S. Wishnok, S.R. Tannenbaum,
Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids, Anal. Biochem. 126
(1982) 131-138.
70. P.F. Pratt, K. Nithipatikom, W.B. Campbell, Simultaneous Determination of Nitrate and
Nitrite in Biological Samples by Multichannel Flow Injection Analysis, Anal. Biochem.
231 (1995) 383-386.

71. H. Habu, I. Yokoi, H. Kabuto, A. Mori, Application of automated flow injection

analysis to determine nitrite and nitrate in mouse brain, Neurochem. 5 (1994) 1571-
1573.

72. I. Yokoi, H. Habu, H. Kabuto, A. Mori, Analysis of nitrite, nitrate, and nitric oxide
synthase activity in brain tissue by automated flow injection technique, Meth. Enzymol.
268 (1996) 152-159.

73. L.A. Ridnour, J.E. Sim, M.A. Hayward, D.A. Wink, S.M. Martin, G.R. Buettner, D.R.
Spitz, A Spectrophotometric Method for the Direct Detection and Quantitation of Nitric
Oxide, Nitrite, and Nitrate in Cell Culture Media, Anal. Biochem. 281 (2000) 223-229.

74. J. Hilton, E. Rigg, Determination of nitrate in lake water by the adaptation of the
hydrazine-copper reduction method for use on a discrete analyser: performance statistics
and an instrument-induced difference from segmented flow conditions, Analyst 108
(1983) 1026-1028.

75. N.A. Zatar, M.A. Abu-Eid, A.F. Eid, Spectrophotometric determination of nitrite and
nitrate using phosphomolybdenum blue complex. Talanta 50 (1999) 819-826.

76. M. Široki, Z. Stefanac, L.J. Marić, Extraction-visible spectrophotometric method for

determination of nitrate: Application to water analysis, Microchem. J. 20 (1975) 483-
491.

77. G. Norwitz, H. Gordon, An improved technique for the spectrophotometric

determination of nitrate with 2,4-xylenol, Anal. Chim. Acta 89 (1977) 177-187.

78. P.W. West, T.P. Ramachandran, Spectrophotometric determination of nitrate using

chromotropic acid. Anal. Chim. Acta 35 (1966) 317-324.
79. K. Horita, M. Satake, Column Preconcentration Analysis–Spectrophotometric
Determination of Nitrate and Nitrite by a Diazotization–Coupling Reaction, Analyst 122
(1997) 1569-1574.

80. G.M. Greenway, S.J. Haswell, P.H. Petsul, Characterisation of a micro-total analytical
system for the determination of nitrite with spectrophotometric detection, Anal. Chim.
Acta 387 (1999) 1-10.

81. M. Barzegar, M.F. Mousavi, A. Nemati, Kinetic spectrophotometric determination of

trace amounts of nitrite by its reaction with molybdosilicic acid blue, Microchem. J. 65
(2000) 159-163.

82. T.A. Doane, W.R. Horwáth, Spectrophotometric Determination of Nitrate with a Single
Reagent, Anal. Lett. 36 (2003) 2713–2722.

83. N. Beda, A. Nedospasov, A spectrophotometric assay for nitrate in an excess of nitrite,

Nitric Oxide 13 (2005) 93-97.

84. J.Z. Zhang, C.J. Fischer, A simplified resorcinol method for direct spectrophotometric
determination of nitrate in seawater. Mar. Chem. 99 (2006) 220-226.

85. M.R. Baezzat, G. Parsaeian, Determination of traces of nitrate in water samples using
spectrophotometric method after its preconcentration on microcrystalline naphthalene.
Quim. Nova 34 (2011) 607-609.

86. B.S. Gentle, P.S. Ellis, M.R. Grace, I.D. McKelvie, Flow analysis methods for the direct
ultra-violet spectrophotometric measurement of nitrate and total nitrogen in freshwaters,
Anal. Chim. Acta 704 (2011) 116-122.

87. N. Chirilă, Comparative Study to Determine the Nitrate ion Using Spectrophotometric
and Chromatographic Methods, Procedia Technol. 12 (2014) 628-631.

88. J. Causse, O. Thomas, A.V. Jung, M.F. Thomas, Direct DOC and nitrate determination
in water using dual pathlength and second derivative UV spectrophotometry, Water
Res. 108 (2017) 312-319.
89. K. Kneipp, H. Kneipp, I. Itzkan, R.R. Dasari, M.S. Feld, Surface-enhanced Raman
scattering and biophysics, J. Phys.: Condens. Matter 14 (2002) R597–R624.

90. M.C. Goldberg, E.R. Weiner, Proc. 4th Annu. Earth Resour. Program Rev., 81 (1972) 1-
14.

91. R. Reeves, Ed., Manual of Remote Sensing, Vol. I, American Society of

Photogrammetry, Falls Church, Va., 1975, pp 1496-1500.

92. Davis, A., M. Bristow, and J. Koningstein, Remote Sensing and Water Resources
Management, Am. Water Res. Assoc., Proc., 17, 239-246 tia7w - I.

93. D.E. Irish, H. Chen, The Application of Raman Spectroscopy to Chemical Analysis,
Appl. Spectrosc. 25 (1971) 1-6.

94. G. Braunlich, G. Gamer, Detection of pollutants in water by Raman spectroscopy,

Water Res. 7 (1973) 1643-1647.

95. E.B. Bradley, C.A. Frenzel, On the exploitation of laser Raman spectroscopy for
detection and identification of molecular water pollutants, Water Res. 4 (1970) 125-128.

96. S.F. Baldwin, C.W. Brown, Detection of ionic water pollutants by laser excited Raman
spectroscopy, Water Res. 6 (1972) 1601-1604.

97. M. Ahmadjian, C.W. Brown, Feasibility of remote detection of water pollutants and oil
slicks by laser-excited Raman spectroscopy, Environ. Sci. Technol. 7 (1973) 452-453.

98. L. V. Haverbeke, C.W. Brown, Water Pollution Studies Using Raman Spectroscopy, in:
Toribara T.Y., Coleman J.R., Dahneke B.E., Feldman I. (eds) Environmental Pollutants.
Environmental Science Research, Vol 13. Springer, Boston, MA, 1978 pp.149-169.

99. W.K. Kowalchyk, P.A. Walker III, M.D. Morris, Rapid Normal Raman Spectroscopy of
Sub-ppm Oxy-Anion Solutions: The Role of Electrophoretic Preconcentration, Appl.
Spectrosc. 49 (1995) 1183-1188.
100. T.H. Kauffmann, M.D. Fontana, Inorganic salts diluted in water probed by Raman
spectrometry: data processing and performance evaluation, J. Sensor. Actuat. B Chem.
209 (2015) 154–161.

101. Z. Li, J. Wang, D. Li, Applications of Raman spectroscopy in detection of water quality,
Appl. Spectrosc. Rev. 51 (2016) 333-357.

102. K.M. Cunningham, M.C. Goldberg, E.R. Weiner, Investigation of Detection Limits for
Solutes in Water Measured by Laser Raman Spectrometry, Anal. Chem. 49 (1977) 70-
75.

103. R.J. Thibeau, L.V. Haverbeke, C.W. Brown, Detection of Water Pollutants by Laser
Excited Resonance Raman Spectroscopy; Pesticides and Fungicides, Appl. Spectrosc.
32 (1978) 98-100.

104. N. Furuya, A. Matsuyuki, S. Higuchi, S. Tanaka, Determination of nitrite ion in waste

and treated waters by resonance Raman spectrometry, Water Res. 14 (1980) 747-752.

105. D.R. Lombardi, C. Wang, B. Sun, A.W. Fountain, T.J. Vickers, C.K. Mann,
Quantitative and qualitative analysis of some inorganic compounds by Raman
spectroscopy, J. Appl. Spectrosc. 48 (1994) 875-883.

106. A. Lanoul, T. Coleman, S.A. Asher, UV Resonance Raman Spectroscopic Detection of

Nitrate and Nitrite in Wastewater Treatment Processes, Anal. Chem. 74 (2002) 1458-
1461.

107. L. He, Y. Liu, M. Lin, J. Awika, D.R. Ledoux, H. Li, A. Mustapha, A new approach
to measure melamine, cyanuric acid, and melamine cyanurate using surface enhanced
Raman spectroscopy coupled with gold nanosubstrates, Sens. Instrum. Food Qual. Saf.
2 (2008) 66–71.

108. S. Nie, S.R. Emory, Probing Single Molecules and Single Nanoparticles by Surface-
Enhanced Raman Scattering, Science 275 (1997) 1102-1106.
109. K. Kneipp, Y. Wang, H. Kneipp, L.T. Perelman, I. Itzkan, R.R. Dasari, Single molecule
detection using surface-enhanced Raman scattering (SERS), Phys. Rev. Lett. 78 (1997)
1667–1670.

110. M. Moskovits, Surface-enhanced Raman spectroscopy: a brief retrospective. J. Raman

Spectrosc. 36 (2005) 485–496.

111. S. Gajaraj, C. Fan, M. Lin, Z. Hu, Quantitative detection of nitrate in water and
wastewater by surface-enhanced Raman spectroscopy, Environ. Monit. Assess. 185
(2013) 5673–5681.

112. P.A. Mosier-Boss, S.H. Lieberman, Detection of Nitrate and Sulfate Anions by Normal
Raman Spectroscopy and SERS of Cationic-Coated, Silver Substrates, Appl. Spectrosc.
54 (2000) 1126-1135.

113. P.A. Mosier-Boss, S.H. Lieberman, Detection of Anions by Normal Raman

Spectroscopy and Surface-Enhanced Raman Spectroscopy of Cationic-Coated
Substrates, Appl. Spectrosc. 57 (2003) 1129- 1137.

114. I. Durickovic, M. Marchetti, Raman spectroscopy as polyvalent alternative for water

pollution detection, IET Sci. Meas. Technol. 8 (2014) 122-128.

115. Y.H. Luo, G.Q. Wen, J.C. Dong, Q.Y. Liu, A.H. Liang, Z.L. Jiang, SERS detection of
trace nitrite ion in aqueous solution based on the nitrosation reaction of rhodamine 6G
molecular probe, Sens. Actuators, B 201 (2014) 336-342.

116. J. Chen, S. Pang, L. He, S.R. Nugen, Highly sensitive and selective detection of nitrite
ions using Fe3O4@SiO2/Au magnetic nanoparticles by surface-enhanced Raman
spectroscopy, Biosens Bioelectron. 85 (2016) 726-733.

117. J.F. Li, Y.F. Huang, Y. Ding, Z.L. Yang, S.B. Li, X.S. Zhou, F.R. Fan, W. Zhang, Z.Y.
Zhou, D.Y. Wu, B. Ren, Z.L. Wang, Z.Q. Tian, Shell-isolated nanoparticle-enhanced
Raman spectroscopy, Nature 464 (2010) 392–395.

118. J.F. Li, S.Y. Ding, Z.L. Yang, M.L. Bai, J.R. Anema, X. Wang, A. Wang, D.Y. Wu, B.
Ren, S.M. Hou, T. Wandlowski, Z.Q. Tian, Extraordinary Enhancement of Raman
 Scattering from Pyridine on Single Crystal Au and Pt Electrodes by Shell-Isolated Au
Nanoparticles, J. Am. Chem. Soc. 133 (2011) 15922–15925.

119. D. Graham, The Next Generation of Advanced Spectroscopy: Surface Enhanced Raman
Scattering from Metal Nanoparticles, Angew Chem. Int. Ed. 49 (2010) 9325–9327.

120. K.G. Zhang, Y.L. Hu, G.K. Li, Diazotization-coupling reaction-based selective
determination of nitrite in complex samples using shell-isolated nanoparticle enhanced
Raman spectroscopy. Talanta 116 (2013) 712–718.

121. P.M. Aker, J. Zhang, W. Nichols, Nitrate ion detection in aerosols using morphology-
dependent stimulated Raman scattering, J. Chem. Phys. 110 (1999) 2202-2207.

122. J.X. Zhang, P.M. Aker, Spectroscopic probing of aerosol particle interfaces, J. Chem.
Phys. 99 (1993) 9366.

123. N. Furuya, A. Matsuyuki, S. Higuchi, S. Tanaka, Determination of nitrate ion in waste

and treated waters by laser raman spectrometry, Water Res. 13 (1979) 371-374.

124. R.J. Magee, S.A.E.F. Shahine, C.L. Wilson, The Determination of Nitrate by Infrared
Spectroscopy, Mikrochim. Acta 6 (1964) 1019-1022.

125. R.T. Yang, M.J.D. Low, Quantitative Analysis of Aqueous Nitrite/Nitrate Solutions by
Infrared Internal Reflectance Spectrometry, Anal. Chem. 45 (1973) 2014-2018.

126. M. Falk, Infrared spectrometric determination of nitrate, nitrite and ammonium ions by
means of matrix isolation in pressed alkali metal halide pellets, Vib. Spectrosc. 1 (1990)
69-79.

127. G. Jiao, S.H. Lips, Nitrate assay by multiple wavelength spectroscopy and analysis of
kinetic parameters of net nitrate uptake by barley seedlings, J. Plant Nutr. 23 (2000) 79-
90.

128. R. Linker, A. Shaviv, Nitrate Determination Using Anion Exchange Membrane and
Mid-Infrared Spectroscopy, Appl. Spectrosc. 60 (2006) 1008-1012.
129. D.J. Goebbert, E. Garand, T. Wende, R. Bergmann, G. Meijer, K.R. Asmis, D.M.
Neumark, Infrared Spectroscopy of the Microhydrated Nitrate Ions NO3-(H2O)1-6, J.
Phys. Chem. A 113 (2009) 7584–7592.

130. J. Huang, P.R. Liu, Q.Y. Sun, H. Zhang, Y. Zhang, K. Wang, Determination of Total
Nitrogen, Ammonia, and Nitrite in River Water by Near-infrared Spectroscopy and
Chemometrics, Anal. Letts. 50 (2017) 1620-1629.

131. J.D.S. Goulden, D. Manning, Infra-red spectroscopy of inorganic materials in aqueous

solution, J. Spectrochim. Acta, Part A 23 (1967) 2249-2254.

132. G.E.B.Y. Ahlijah, E.F. Mooney, The attenuated total reflection spectra of polyatomic
inorganic anions-II.: The nitrogen containing anions, Spectrochi. Acta, Part A 25 (1969)
619-627.

133. E.J. Elzinga, D. Peak, D.L. Sparks, Spectroscopic studies of Pb(II)-sulfate interactions
at the goethite-water interface, Geochim. Cosmochim. Acta 65 (2001) 2219-2230.

134. S.H. Strauss, M.A. Odom, G.N. Herbert, B.J. Clapsaddle, ATR-FTIR Detection of >/=
25 μg/L Aqueous Cyanide, Perchlorate, and PFOS (PDF), J. Am. Water Works Assoc.
94 (2002) 109-115.

135. A. Shaviv, A. Kenny, I. Shmulevitch, L. Singher, Y. Raichlin, A. Katzir, Direct

Monitoring of Soil and Water Nitrate by FTIR Based FEWS or Membrane Systems,
Environ. Sci. Technol. 37 (2003) 2807-2812.

136. R. Linker, A. Kenny, A. Shaviv, L. Singher, I. Shmulevich, Fourier Transform Infrared–

Attenuated Total Reflection Nitrate Determination of Soil Pastes Using Principal
Component Regression, Partial Least Squares, and Cross-Correlation, Appl. Spectrosc.
58 (2004) 516-520.

137. B.R. Jahn, R. Linker, S.K. Upadhyaya, A. Shaviv, D.C. Slaughter, I. Shmulevich, Mid-
infrared Spectroscopic Determination of Soil Nitrate Content, Biosyst. Eng. 94 (2006)
505–515.
138. E. Choe, F. van der Meer, D. Rossiter, C. van der Salm, K.W. Kim, An Alternate
Method for Fourier Transform Infrared (FTIR) Spectroscopic Determination of Soil
Nitrate Using Derivative Analysis and Sample Treatments, Water Air Soil Pollut. 206
(2010) 129–137.

139. Y.Q. Shao, C.W. Du, Y.Z. Shen, F. Ma, J.M. Zhou, Rapid Determination of N Isotope
Labeled Nitrate Using Fourier Transform Infrared Attenuated Total Reflection
Spectroscopy, Chin. J. Anal. Chem. 42 (2014) 747–752.

140. Y. Shao, C. Du, J. Zhou, F. Ma, Y. Zhu, K. Yang, C. Tian, Quantitative analysis of
different nitrogen isotope labelled nitrates in paddy soil using mid-infrared attenuated
total reflectance spectroscopy, Anal. Methods 9 (2017) 5388-5394.

141. T. Kumamaru, E. Tao, N. Okamoto, Y. Yamamoto, A new method for the

determination of nitrate ions by atomic absorption spectrophotometry, Bull. Chem. Soc.
Jpn. 38 (1965) 2204-2204.

142. M.E. Houser, M.I. Fauth, Indirect Determination of Nitrate, Nitrite, and Nitro Groups
by Atomic Absorption Spectrophotometry, Microchem. J. 15 (1970) 399-408.

143. Y. Yamamoto, T. Kumamaru, Y. Hayashi, Y. Otani, Atomic absorptiometric

determination of nitrate ion by solvent extraction of the ion-pair of bis-
(neocuproine)copper(I) nitrate, Bunseki Kagaku 18 (1969) 359-363.

144. M. Gallego, M. Silva, M. Valcarcel, Determination of nitrate and nitrite by continuous

liquid-liquid extraction with a flow-injection atomic-absorption detection system, Fres.
Z. Anal. Chem. 323 (1986) 50-53.

145. M. Silva, M. Gallego, M. Valcarcel, Sequential atomic absorption spectrometric

determination of nitrate and nitrite in meats by liquid-liquid extraction and a flow
injection system, Anal. Chim. Acta. 179 (1986) 341-349.

146. S.S.M. Hassan, New atomic-absorption spectrometric, potentiometric and polarographic

methods for the determination of nitrates, Talanta 28 (1981) 89-93.
147. E.V. Alonso, M.T.S. Cordero, M.P.C. Torres, A.G. Torres, J.M.C. Pavon, Indirect
determination of nitrate by electrothermal atomic absorption spectrometry using an on-
line cadmium microcolumn, Analyst, 123 (1998) 1561–1564.

148. J. Anwar, T. Haque, Indirect Atomic Absorption Spectrophotometric Method For the
Determination of Nitrate, J. Chem. Soc. Pak. 7 (1985) 209-213.

149. J. Anwar, Riaz-ur-Rahman, M.I. Farooqi, M. Yousaf, Indirect determination of nitrate

by atomic absorption spectrophotometry, Pak. J. Sci. Ind. Res. 29 (1986) 332-337.

150. S. Motomizu, H. Mikasa, K. Toei, Fluorimetric determination of nitrate in natural

waters with 3-amino-l,5-naphthelenedisulphonic acid in a flow-injection system, Anal.
Chim. Acta. 193 (1987) 343-347.

151. L.J. Dombrowsky, E.J. Pratt, Fluorimetric method for determining nanogram quantities
of nitrite ion. Anal. Chem. 44 (1972) 2268-2272.

152. I. Hornyak, L. Szekelyhidi, Spectrofluorimetric method for determining trace quantities

of nitrite ion. Mikrochim. Acta 2 (1983) 355-359.

153. T. Ohta, Y. Arai, S. Takitani, Fluorometric determination of nitrite with 4-

hydroxycoumarin, Anal. Chem. 58 (1986) 3132-3135.

154. S.H. Lee, L.R. Field, Postcolumn fluorescence detection of nitrite, nitrate, thiosulfate,
and iodide anions in high-performance liquid chromatography, Anal. Chem. 56 (1984)
2647-2653.

155. D.B. Gladilovich, N.N. Grigorev, K. Sedyk, N.E. Pozdnyakova, Fluorimetric and
extraction-fluorimetric determinations of nitrites with 2,3-diaminonaphthelene, J. Anal.
Chem. of USSR 44 (1989) 1122-1126.

156. M.A.T. Fernández-Argüelles, B. Cañabate, J.M. Costa-Fernández, R. Pereiro, A. Sanz-

Medel, Flow injection determination of nitrite by fluorescence quenching, Talanta 62
(2004) 991-995.
157. R. Lumpp, J. Reichert, H.J. Ache, An optical sensor for the detection of nitrate, Sens.
Actuators, B 7 (1992) 473-475.

158. H. Liu, G. Yang, E. Abdel-Halim, J.J. Zhu, Highly selective and ultrasensitive detection
of nitrite based on fluorescent gold nanoclusters, Talanta 104 (2013) 135-139.

159. M. Nakamura, Resorcinol as Fluorimetric Reagent for the Determination of Nitrate,

Anal. Lett. 13 (1980) 771-779.

160. S. Biswas, B. Chowdhury, B.C. Ray, A novel spectrofluorimetric method for the ultra
trace analysis of nitrite and nitrate in aqueous medium and its application to air, water,
soil and forensic samples, Talanta 64 (2004) 308-312.

161. Q.H. Liu, X.L. Yan, J.C. Guo, D.H. Wang, L. Li, F.-Y. Yan, L.-G. Chen,
Spectrofluorimetric determination of trace nitrite with a novel fluorescent probe,
Spectrochim. Acta, Part A 73 (2009) 789-793.

162. X. Zhang, H. Wang, N.N. Fu, H.S. Zhang, A fluorescence quenching method for the
determination of nitrite with Rhodamine 110, Spectrochim. Acta, Part A 59 (2003)
1667-1672.

163. F. Gao, L. Zhang, L. Wang, S. She, C. Zhu, Ultrasensitive and selective determination
of trace amounts of nitrite ion with a novel fluorescence probe mono [6-N (2-carboxy-
phenyl)]-β-cyclodextrin, Anal. Chim. Acta 533 (2005) 25-29.

164. X. Wang, E. Adams, A. Van Schepdael, A fast and sensitive method for the
determination of nitrite in human plasma by capillary electrophoresis with fluorescence
detection, Talanta 97 (2012) 142-144.

165. H.D. Axelrod, N.A. Engel, Fluorometric determination of subnanogram levels of nitrite
using 5-aminofluorescein, Anal. Chem. 47 (1975) 922-924.

166. A. Lapat, L. Szekelyhidi, I. Hornyak, Spectrofluorimetric determination of 1,3,5-

trinitro-1,3,5-triazacyclohexane (hexogen, RDX) as a nitramine type explosive, Biomed.
Chromatogr. 11 (1997) 102-104.
167. A. Buldt, U. Karst, Determination of Nitrite in Waters by Microplate Fluorescence
Spectroscopy and HPLC with Fluorescence Detection, Anal. Chem. 71 (1999) 3003-
3007.

168. N.Q. Jie, D.L. Yang, Q.B. Jiang, Q. Zhang, L. Wei, A Fluorescence Quenching Method
for the Determination of Nitrite with Indole, Microchem. J. 62 (1999) 371-376.

169. Z. Huang, T. Korenaga, M.I.H. Helaleh, Kinetic Spectrofluorimetric Determination of

Nitrite in Water Samples and Nitrogen Dioxide in the Atmosphere Sampled by the
Liquid Droplet Method, Mikrochim. Acta 134 (2000) 179-183.

170. H. Wang, W. Yang, S.C. Liang, Z.M. Zhang, H.S. Zhang, Spectrofluorimetric
determination of nitrite with 5,6-diamino-1,3-naphthalene disulfonic acid, Anal. Chim.
Acta 419 (2000) 169-173.

171. M.I.H. Helaleh, T. Korenaga, Fluorometric determination of nitrite with acetaminophen,

Microchem. J. 64 (2000) 241-246.

172. K. Geetha, N. Balasubramanian, An Extractive Spectrophotometric Method for the

Determination of Nitrite Using Fluorescein Amine Isomer I, Anal. Lett. 33 (2000) 1869-
1883.

173. Y.X. Guo, Q.F. Zhang, X. Shangguang, G. Zhen, Spectrofluorimetric determination of

trace nitrite with o-phenylenediamine enhanced by hydroxypropyl-β-cyclodextrin,
Spectrochim. Acta, Part A 101 (2013) 107-111.

174. H. Li, C.J. Meininger, G.Y. Wu, Rapid determination of nitrite by reversed-phase high-
performance liquid chromatography with fluorescence detection, J. Chromatogr. B:
Biomed. Sci. Appl. 746 (2000) 199-207.

175. R.T. Masserini Jr., K.A. Fanning, A sensor package for the simultaneous determination
of nanomolar concentrations of nitrite, nitrate, and ammonia in seawater by
fluorescence detection, Mar. Chem. 68 (2000) 323–333.
176. S.I. Fujii, T. Tokuyama, M. Abo, A. Okubo, Fluorometric determination of sulfite and
nitrite in aqueous samples using a novel detection unit of a microfluidic device, Anal.
Sci. 20 (2004) 209-212.

177. M. Martínez-Tomé, R. Esquembre, R. Mallavia, C. Mateo, Immobilization and

Characterization of 2, 3-diaminonaphthalene/cyclodextrin Complexes in a Sol–Gel
Matrix: A New Fluorimetric Sensor for Nitrite, J. Fluoresc. 19 (2009) 119-125.

178. K.J. Huang, W.Z. Xie, H.S. Zhang, H. Wang, Ultra-trace level determination of nitrite
in human saliva by spectrofluorimetry using 1,3,5,7-tetramethyl-8-(3, 4-
diaminophenyl)-difluoroboradiaza-s-indacene, Microchim. Acta 161 (2008) 201-207.

179. K.N. Lin, M.A. Jian, D.X. Yuan, Y.M. Huang, S.C. Feng, Q.L. Wu, Determination of
Nitrate in Seawater with Valve-Free Continuous Flow Analysis, Chin. J. Anal. Chem.
45 (2017) 151-156.

180. X.Q. Zhan, D.H. Li, H. Zheng, J.G. Xu, A sensitive fluorimetric method for the
determination of nitrite and nitrate in seawater by a novel red-region fluorescence dye,
Anal. Lett. 34 (2001) 2761-2770.

181. R. Li, J.C. Yu, Z. Jiang, R. Zhou, H. Liu, A solid-phase fluorescent quenching method
for the determination of trace amounts of nitrite in foods with neutral red, J. Food Drug
Anal. 11 (2003) 251-257.

182. T. Chen, A. Tong, Y. Zhou, 2-Amino-5, 7-dimethyl-1, 8-naphthyridine as a fluorescent

reagent for the determination of nitrite, Spectrochim. Acta, Part A 66 (2007) 586-589.

183. M. Li, H. Wang, X. Zhang, H.S. Zhang, Development of a new fluorescent probe: 1, 3,
5, 7-tetramethyl-8-(4CE-aminophenyl)-4, 4-difluoro-4-bora-3a, 4a-diaza-s-indacence
for the determination of trace nitrite, Spectrochim. Acta, Part A 60 (2004) 987-993.

184. K.J., Huang, H. Wang, Y.H. Guo, R.L. Fan, H.S. Zhang, Spectrofluorimetric
determination of trace nitrite in food products with a new fluorescent probe 1,3,5,7-
tetramethyl-2,6-dicarbethoxy-8-(3′,4′-diaminophenyl)-difluoroboradiaza-s-indacene,
Talanta, 69 (2006) 73-78.
185. L. Lu, C. Chen, D. Zhao, F. Yang, X. Yang, A simple and sensitive assay for the
determination of nitrite using folic acid as the fluorescent probe, Anal. Methods 7
(2015) 1543-1548.

186. K. Yoshizumi, K. Aoki, Determination of nitrate by a flow system with a

chemiluminescent nitrogen oxide (NOx) analyzer, Anal. Chem. 57 (1985) 737-740.

187. C. Garside, A chemiluminescent technique for the determination of nanornolar

concentrations of nitrate and nitrite in seawater, Mar. Chem. 11 (1982) 159-167.

188. C. Garside, The vertical distribution of nitrate in open ocean surface water, Deep-Sea
Res. 32 (1985) 723-732.

189. R. Braman, S.A. Hendrix Nanogram nitrite and nitrate determination in environmental
and biological materials by vanadium(III) reduction with chemiluminescence detection.
Anal. Chem. 61 (1989) 2715–2718.

190. Walters, C.L., R.J. Hart, M.J. Downs, S. Perse, and P.L.R.Z. Smith, Determination of
Nitrite at Low Level Without Prior Extraction, Lebensm.Unters.-Forsch. 1978,167, 229-
232.

191. R.C. Doerr, J.B. Fox, Jr., L. Lakritz, W. Fiddler, Determination of nitrite in cured meats
by chemiluminescence detection, Anal. Chem. 53 (1981) 381-384.

192. F. Yang, E. Troncy, M. Francoeur, B. Vinet, P. Vinay, G. Czaika, G. Blaise, Effects of

reducing reagents and temperature on conversion of nitrite and nitrate to nitric oxide
and detection of NO by chemiluminescence, Clin. Chem. 43 (1997) 657-662.

193. J.J. Lingane, R.L. Pecsok, Volumetric determination of nitrate ion by reduction with
chromous ion, Anal. Chem. 21 (1949) 622-625.

194. M.C. Cresser, Nitrate determination by reduction to ammonia and gas-phase ultraviolet
absorption spectrometry. Analyst 102 (1977) 99–103.

195. R.D. Cox, C.W. Frank, Determination of nitrate and nitrite in blood and urine by
chemiluminescence, J. Anal. Toxicol. 6 (1982) 148-152.
196. E. Nagababu, J.M. Rifkind, Measurement of plasma nitrite by chemiluminescence
without interference of S-, N-nitroso and nitrated species, Free Radical Biol. Med. 42
(2007) 1146-1154.

197. T. Aoki, S. Fukuda, Y. Hosoi, H. Mukai, Rapid flow injection analysis method for
successive determination of ammonia, nitrite, and nitrate in water by gas-phase
chemiluminescence, Anal. Chim. Acta 349 (1997) 11-16.

198. Y. Kanda, M. Taira, Flow-injection analysis method for the determination of nitrite and
nitrate in natural water samples using a chemiluminescence NOx monitor, Anal. Sci. 19
(2003) 695-699.

199. N.P. Sen, P.A. Baddoo, S.W. Seaman, Rapid and sensitive determination of nitrite in
foods and biological materials by flow injection or high-performance liquid
chromatography with chemiluminescence detection, J. Chromatogr. 673 (1994) 77–84.

200. T. Aoki, Continuous flow determination of nitrite with membrane

separation/chemiluminescence detection, Biomed. Chromatogr. 4 (1990) 128-130.

201. T. Aoki, M. Wakabayashi, Simultaneous flow injection determination of nitrate and

nitrite in water by gas-phase chemiluminescence, Anal. Chim. Acta 308 (1995) 308-
312.

202. A.J. Dunham, R.M. Barkley, R.E. Sievers, Aqueous Nitrite Ion Determination by
Selective Reduction and Gas Phase Nitric Oxide Chemiluminescence, Anal. Chem. 67
(1995) 220-224.

203. A. Townshend, Solution chemiluminescence-some recent analytical developments.

Plenary lecture, Analyst, 115 (1990) 495-500.

204. M.C. Icardo, J.G. Mateo, J.M. Calatayud, Determination of nitrite by inhibition of the
chemiluminescence of acriflavine in a flow-injection assembly, Analyst 126 (2001)
1423-1427.

205. P. Mikuska, Z. Vecera, Simultaneous determination of nitrite and nitrate in water by

chemiluminescent flow-injection analysis, Anal. Chim. acta 495 (2003) 225-232.
206. Q. Yue, Z. Song, Assay of femtogram level nitrite in human urine using
luminol.myoglobin chemiluminescence, Microchem. J. 84 (2006) 10-13.

207. D. He, Z. Zhang, Y. Huang, Y. Hu, Chemiluminescence microflow injection analysis

system on a chip for the determination of nitrite in food, Food Chem. 101 (2007) 667-
672.

208. M. Yaqoob, B. Folgado Biot, A. Nabi, P.J. Worsfold, Determination of nitrate and
nitrite in freshwaters using flow injection with luminol chemiluminescence detection,
Luminescence 27 (2012) 419-425.

209. A. Mitschke, F.M. Gutzki, D. Tsikas, 9- and 10-Nitro-oleic acid do not interfere with
the GC–MS quantitative determination of nitrite and nitrate in biological fluids when
measured as their pentafluorobenzyl derivatives, J. Chromatogr. B 851 (2007) 287-291.

210. S.M. Helmke, M.W. Duncan, Measurement of the NO metabolites, nitrite and nitrate, in
human biological fluids by GC–MS, J. Chromatogr. B 851 (2006) 83-92.

211. D. Tsikas, Simultaneous Derivatization and Quantification of the Nitric Oxide

Metabolites Nitrite and Nitrate in Biological Fluids by Gas Chromatography/Mass
Spectrometry, Anal. Chem. 72 (2000) 4064-4072.

212. S. Kage, K. Kudo, N. Ikeda, Determination of nitrate in blood by gas chromatography

and gas chromatography-mass spectrometry, J. Chromatogr. B 742 (2000) 363-368.

213. S. Kage, K. Kudo, N. Ikeda, Simultaneous Determination of Nitrate and Nitrite in

Human Plasma by Gas Chromatography-Mass Spectrometry, J. Anal. Toxicol. 26
(2002) 320-324.

214. M. Akyuz, S. Ata, Determination of low level nitrite and nitrate in biological, food and
environmental samples by gas chromatography–mass spectrometry and liquid
chromatography with fluorescence detection, Talanta 79 (2009) 900–904.

215. B. Campanella, M. Onor, E. Pagliano, Rapid determination of nitrate in vegetables by

gas chromatography mass spectrometry, Anal. Chim. Acta 980 (2012) 33-40.
216. M. Honma, C.L. Smith, Quantitative Analysis of Organic Nitrogen by Flame
Spectroscopy, Anal. Chem. 26 (1954) 458-462.

217. R. Belcher, S.L. Bogdanski, A.C. Calokerinos, A. Townshend, Determination of

ammonium- and nitrate-nitrogen by molecular emission cavity analysis, Analyst 106
(1981) 625-635.

218. I.Z. Al-Zamil, A. Townshend, Molecular emission cavity analysis: Part 23.
Determination of Nitrite and Nitrate after Conversion to Nitrogen Monoxide, Anal.
Chim. Acta 142 (1982) 151-157.

219. A. Celik, E. Henden, Determination of ammonium-, nitrite- and nitrate-nitrogen by

molecular emission cavity analysis using a cavity containing an entire flame, Analyst
114 (1989) 563-566.

220. R. Linker, Determination of Nitrate Concentration in Soil via Photoacoustic

Spectroscopy Analysis of Ion Exchange Membranes, Appl. Spectrosc. 62 (2008) 248-
250.

221. P. Qian, J.J. Schoenau, Use of Ion-Exchange Membrane to Assess Nitrogen-Supply

Power of Soils, J. Plant Nutr. 28 (2005) 2193-2200.

222. J.E. Szillery, I.J. Fernandez, S.A. Norton, L.E. Rustad, A.S. White, Using Ion-
Exchange Resins to Study Soil Response to Experimental Watershed Acidification,
Environ. Monit. Assess. 116 (2006) 383-398.

223. M.J. Pawe, E.F.J. Woods, P. Gibbs, R. Amma, Electron paramagnetic resonance
spectroscopic investigation of the inhibition of the phosphorwlastic system of
Clostridum sporogenes by nitrite, J. Gen. Microbiol. 136 (1990) 2067-2076.

224. A. Wennmalm, A.S. Petersson, Analysis of nitrite as a marker for endothelium-derived

relaxing factor in biological fluids using electron paramagnetic resonance spectrometry,
J. Cardiovasc. Pharmacol. 17 (1991) S34-S40.
225. N.D. Yordanov, E. Novakova, S. Lubenova, Consecutive estimation of nitrate and
nitrite ions in vegetables and fruits by electron paramagnetic resonance spectrometry,
Anal. Chim. Acta 437 (2001) 131-138.

226. B.I. Goetz, H.W. Shields, S. Basu, P. Wang, S.B. King, N. Hogg, M.T. Gladwin, D.B.
Kim-Shapiro, An electron paramagnetic resonance study of the affinity of nitrite for
methemoglobin, Nitric Oxide 22 (2010) 149-54.

227. D.E. Schwab, J.S. Stamler, D.J. Singel, EPR Spectroscopy of Nitrite Complexes of
Methemoglobin, Inorg. Chem. 49 (2010) 6330–6337.

228. M. Bawn, F. MacMillan, Electron Paramagnetic Resonance Investigation of Nitrite

Binding in Myoglobin, bxv_biochem (2018) doi: org/10.1101/252775.

229. J. Lu, A.C. Chang, L. Wu, Distinguishing sources of groundwater nitrate by 1H NMR
of dissolved organic matter, Environ. Pollut. 132 (2004) 365-374.
 GRAPHICAL ABSTRACT

HIGHLIGHTS

 Spectroscopic methods for detection of nitrate and nitrite have been reviewed.
 Their basic principle, advantages and disadvantages have been mentioned.
 Detection limits, detection range, RSD%, sample throughput/h have been tabulated.
 Literature survey of more than 200 research papers has been presented.