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Physico-chemical standardization of Sitopaladi churna

Article in Ancient Science of Life · March 2012

DOI: 10.4103/0257-7941.103187 · Source: PubMed

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Original Article

Physico-chemical standardization of
Sitopaladi churna
Inder Kumar Makhija, Chandrashekara Shastry Shreedhara, Holavana Hally Nanjundaiah Setty Aswatha Ram
Department of Pharmacognosy, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, Karnataka, India

the standards mentioned in Ayurveda. Most of the tests

ABSTRACT:
mentioned in ancient literature appear to be based on
Background: Standardization of a compound Ayurvedic
observation and seem subjective without valid scientific
formulation is a critical and essential issue to be considered in
backing; therefore, formulation prepared may not have
assuring the therapeutic efficacy and safety and to rationalize their
use in the health care. Sitopaladi churna is a reputed polyherbal the desired quality and batch-to-batch consistency.
formulation of Ayurveda. It is prescribed for the treatment of Quality is the critical determinant of safety and efficacy
pleurodynia, intercostal neuralgia, cold, cough associated with of herbal medicines; however, herbal formulations rarely
bronchitis, pneumonia, tuberculosis, viral respiratory infection, meet the standards of quality. Hence, there is a need for
and in pharyngeal and chest congestion. standardization, and development of reliable quality
Objective: The present study aimed at physico-chemical protocols for Ayurvedic formulations using modern
standardization of in-house and two marketed brands of Sitopaladi techniques of analysis is extremely important. [2,3] The
churna. World Health Organization (WHO) has appreciated the
Materials and Methods: In our investigation, in-house churna and importance of medicinal plants for public health care
two commercial brands of Sitopaladi churna were standardized in developing nations and has evolved guidelines to
based on powder microscopy, physico-chemical evaluations, support the member states in their efforts to formulate
thin layer chromatography (TLC) and high performance thin layer national policies on traditional medicine and to study
chromatography (HPTLC) finger printing as per standard procedures. their potential usefulness including evaluation, safety, and
Results: The set parameters were sufficient to evaluate the churna efficacy.[4-6] The present study deals with Sitopaladi churna
based on various physico-chemical parameters. (SPC), a polyherbal Ayurvedic formulation prescribed for
Conclusion: The data evolved can be adopted for laying down pleurodynia, intercostal neuralgia, cold, cough associated
the standards for the manufacturing units of Sitopaladi churna. with bronchitis, pneumonia, tuberculosis, burning
Key words: High performance thin layer chromatography, sensation in extremities, supportive agent for allergy,
physico-chemical, Sitopaladi churna, standardization viral respiratory infection, digestive impairment, and in
pharyngeal and chest congestion. [7,8] The investigation
was carried out to develop standardization parameters.
The objectives include performing powder microscopic
characterization, physico-chemical analysis, and thin
INTRODUCTION layer chromatography (TLC) and high performance thin
Ayurveda, the traditional Indian medicine, is the “great layer chromatography (HPTLC) fingerprint profile for the
tradition” with sound philosophical, experiential, and quantification of piperine and cinnamaldehyde [Figure 1]
experimental basis. The Ayurvedic system touted as an in SPC samples.
“alternative system of medicine” has already gained
worldwide attention due to increased side effects of drugs, The two standards quantified by HPTLC have been
lack of remedy for several chronic diseases, microbial reported to possess significant biological activities. Piperine
resistance, high cost of synthetic drugs, and emerging
diseases. These are some facts for renewed public interest Access this article online
in traditional medicines. With increasing demand for safer Quick Response Code:
Website:
drugs, attention has been drawn to the quality, safety,
www.ancientscienceoflife.org
efficacy, and standards of the Ayurvedic formulations.[1]
DOI:
Ayurvedic pharmacy advocates the use of quality control 10.4103/0257-7941.103187
tests to make sure that the formulated products adhere to
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Makija, et al.: Standardization of Sitopaladi churna

Table 1: Ingredients of Siwtopaladi churna

Sanskrit name Plant name Part used Quantity
Sitopala Saccharum Sugar candy 192 parts
officinarum
Vamsalochan Bambusa Siliceous 96 parts
arundinacea concretion
Pippali Piper longum Dried fruit 48 parts
Ela Elettaria Dried seed 24 parts
cardamomum
Twak Cinnamonum Stem bark 12 parts
Figure 1: Molecular structure of piperine and cinnamaldehyde zeylanicumm

is reported to have as an antidepressant, hepatoprotective, total ash, water -soluble ash, acid-insoluble ash, water-
anti-metastatic, antithyroid, immunomodulatory, soluble extractives, alcohol-soluble extractives, foaming
a n t i t u m o r, [ 9 ] a n t i p l a t e l e t , [ 1 0 ] a n t i o x i d a n t , [ 11 ] a n d index, loss on drying, and pH (10% aqueous solution) as
antiamoebic[12] activities. Cinnamaldehyde is reported per standard techniques.[21] Micromeritic characteristics
to show antidiabetic,[13,14] antifungal,[15] antibacterial,[16] like bulk density, tap density, angle of repose, Haussner
anticancer,[17] antimutagenic,[18] and anti-inflammatory[19] ratio and Carr’s index were determined for SPC samples.
activities. [22,23]
Aqueous and methanol extracts of SPC samples
prepared by hot extraction were used for screening of
constituents like alkaloids, glycosides, flavonoids, tannins,
MATERIALS AND METHODS sterols, terpenes, fixed oil, resin, protein, and gums.[20] Total
percentage of reducing sugar, tannins, and flavonoids of
Collection and identification of plant materials
SPC samples was determined.[24-26] Fluorescence analysis
The raw drugs used in the in-house SPC-I formulation were was carried out as per the method of Chase and Pratt.[27]
procured from the local market of Udupi, Karnataka, India, SPC samples were analyzed for presence of heavy metals
and authenticated by botanist Dr. K. Gopal Krishna Bhat, like lead (Pb), arsenic (As), cadmium (Cd), and mercury
Professor, Department of Botany, Poorna Prajna College, (Hg) by atomic absorption spectroscopy (AA 240, Varian,
Udupi, Karnataka. A voucher specimen of the same was The Netherlands).[28]
deposited in the museum of Department of Pharmacognosy,
Manipal College of Pharmaceutical Sciences. Commercially
Quantification of piperine and cinnamaldehyde by HPTLC
available brands of SPC [Baidynath Ayurved Bhawan Pvt.
Ltd., Kolkata, India (SPC-II) and Dabur India Ltd., New densitometry
Delhi, India (SPC-III)] were procured from local market. TLC conditions
SPC-I was prepared according to Ayurvedic Formulary of Silica gel 60 F254 TLC plates with aluminum sheet support
India by mixing equal parts by weight of each of the five (0.2 mm thickness) (E. Merck) were used. The syringe was
ingredients of formulation [Table 1].[7] a 100 µL one (Hamilton) and spotting device used was
Camag Linomat V spotter (Camag, Muttenz, Switzerland).
Chemicals The developing chamber was a Camag glass trough
chamber (20 × 10 cm) previously saturated with mobile
All the solvents and chemicals of analytical grade were
phase vapor for 30 min. Densitometry was performed with
purchased from E. Merck and S. D. Fine Chemicals,
a Camag TLC Scanner 3 and Camag Reprostar 3 linked
Mumbai. Piperine (purity 97%) and cinnamaldehyde (purity
to Wincats software (V 3.15, Camag). The experimental
98%) were purchased from Sigma-Aldrich, Bangalore, India.
conditions were temperature of 25 ± 2°C and relative
humidity of 40%.
Physico-chemical evaluations
Organoleptic parameters such as varna (color), gandha Preparation of standard solution
(odor), ruchi (taste), aakruti (shape), and parimana (size) Piperine:
were analyzed and recorded. Powder microscopy of Stock solution of 1000 µg/mL of piperine was prepared
shade-dried powder was carried out using Olympus in methanol. Aliquots (0.1–1 mL) of stock solution were
BX 41 microscope.[20] Physico-chemical characteristics of transferred to 10 mL volumetric flasks and the volume of
SPC samples were analyzed by quantitative analysis for each was adjusted to 10 mL with methanol, so as to obtain

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Makija, et al.: Standardization of Sitopaladi churna

standard solutions containing 10, 20, 30, 40, 50, 60, 70, 80, SPC-I, SPC-II, and SPC-III was spotted in duplicate on
90, and 100 µg/mL of piperine, respectively. a TLC plate using an automated Linomat V applicator.
The plates were developed, scanned, and peak areas
Cinnamaldehyde: and absorption spectra were recorded. The amounts of
Stock solution of 100 µg/mL of cinnamaldehyde was prepared piperine and cinnamaldehyde were calculated using their
in methanol. Aliquots (0.1–1 mL) of stock solution were respective calibration curves. The identity and purity
transferred to 10 mL volumetric flasks and the volume of of the bands of piperine and cinnamaldehyde in the
each was adjusted to 10 mL with methanol, so as to obtain sample extract track were checked by overlaying their
standard solutions containing 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 µg/ UV absorption spectra at start, middle, and end positions
mL of cinnamaldehyde, respectively. of the band.

Preparation of sample solution Method validation

One hundred milligrams of methanolic extract of each
International Conference on Harmonisation guidelines
of SPC-I, SPC-II, and SPC-III dissolved in 10 mL of
were followed for the validation of the analytical methods
methanol was used for quantification of piperine and
developed for precision, repeatability, and accuracy.[29,30]
cinnamaldehyde.
Instrumental precision was checked by repeated scanning
(n = 7) of the same spot of piperine (500 ng/spot) and
Preparation of calibration curves cinnamaldehyde (50 ng/spot) expressed as relative standard
Piperine: deviation (%RSD). The repeatability of the method was
10 µl of each of the standard solutions of piperine (100–
confirmed by analyzing 500 ng/spot of piperine and 50 ng/
1000 ng/spot) was spotted (band width: 6 mm) in triplicate
spot of cinnamaldehyde individually on a TLC plate (n = 5)
on a TLC plate using an automated Linomat V applicator.
and expressed as %RSD. The inter-day and intra-day
The plates were developed in a twin trough chamber
variation of the method was studied by analyzing aliquots
(20 × 10 cm) up to a distance of 9 cm using a mobile phase
of standard solution containing 300, 500, and 700 ng/spot
of toluene:ethyl acetate:formic acid (5:3.5:0.5 v/v/v) (12 mL)
of piperine and 30, 50, and 70 ng/spot cinnamaldehyde
at a temperature of 25 ± 2°C and 40% relative humidity.
on the same day and on different days and the results
The plate was air dried, and scanned at 342 nm in the
were expressed as %RSD. For the evaluation of limit of
absorbance-reflectance mode using a deuterium lamp.
detection (LOD) and limit of quantification (LOQ), different
Calibration curve of piperine was obtained by plotting
concentrations of the standard solutions of piperine and
peak area versus applied concentration of piperine. Video
cinnamaldehyde were applied along with methanol as
densitometry of the chromatogram was carried out with
the help of Camag Reprostar 3. blank and they were determined on the basis of signal-
to-noise (S/N) ratio. LOD was measured at an S/N of 3:1
Cinnamaldehyde: and LOQ at an S/N of 10:1. The specificity of the method
10 µl of each of the standard solutions of cinnamaldehyde was measured as per Bhandari et al. [31] The accuracy of
(10–100 ng/spot) was spotted (band width: 6 mm, distance the method was assessed by performing recovery study
between the tracks: 12 mm) in triplicate on a TLC plate at three different levels (50, 100, and 150% addition of
using an automated Linomat V applicator. The plates were piperine and cinnamaldehyde). The percentage recovery
developed in a twin trough chamber up to a distance of 9 cm and the average percentage recovery for each standard
using an optimized mobile phase of toluene:chloroform were calculated. Specificity was ascertained by analyzing
(8:2 v/v) (12 mL) at a temperature of 25 ± 2°C and 40% relative reference compounds and samples. The bands for piperine
humidity. The plate was air dried, and scanned at 310 nm and cinnamaldehyde from sample solutions were confirmed
in the absorbance-reflectance mode using a deuterium by comparing the Rf and spectra of the bands to those of
lamp. Calibration curve of cinnamaldehyde was obtained the standards.
by plotting peak area versus applied concentration of
cinnamaldehyde. Video densitometry of the chromatogram
was carried out with the help of Camag Reprostar 3. RESULTS AND DISCUSSION
Physico-chemical evaluation
Quantification of piperine and cinnamaldehyde in the
The samples of SPC were found to be brown-colored,
SPC samples moderately fine powder, with a pleasant smell and spicy
Ten microliters of each of the methanolic extracts of taste. All the samples passed through 60 mesh size and not

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Makija, et al.: Standardization of Sitopaladi churna

less than 50% passed through 85 mesh size. Microscopic methanolic extracts of SPC samples were studied
characterization [Figures 2a-i] revealed the presence of spectrophotometrically for spectrum measurement, and
big siliceous crystals (Bambusa arundinacea); epidermis of colorimetric analyses like total carbohydrate sugar by
testa was composed of yellow-brown prosenchymatous phenol sulfuric acid reagent, total tannin by Folin–Denis
cells with pitted walls, and globules of volatile oil with reagent, and total flavonoid by aluminum trichloride
underlying hypodermis and epidermis were present reagent were carried out [Table 4]. Fluorescence analysis
(Elettaria cardamomum); phloem parenchyma was was carried out to check the chemical nature of drug with
associated with occasional phloem fiber and big isolated different reagents; data are depicted in Table 5. Heavy
oil cell, and group of lignified cork cells was associated metal contents of SPC samples were found to be within
with pericyclic fiber, tannin contents (Cinnamonum permissible limits, except Pb in SPC-I [Table 6].
zeylanicumm); stone cell with a broad lumen was present
and group of spiral vessels formed a vascular strand and
perisperm cells (Piper longum). Quantitative physico-
TLC fingerprint and co-chromatography
chemical analysis for the SPC samples was performed TLC fingerprint profile is a systematic representation
for the parameters like total ash, water-soluble ash, acid- of all the constituents of sample resolved in the given
insoluble ash, water-soluble extractives, alcohol-soluble chromatographic system. It gives a semi-quantitative
extractives, loss on drying, and pH [Table 2]. Micromeritic sketch of the chemical profile of the sample. In the present
parameters of SPC samples were also analyzed and the work, we developed simple, convenient, and time-saving
data are depicted in Table 2. Phytochemical constituents TLC methods for co-chromatography with two marker
like alkaloid, carbohydrates, flavonoid, tannins, saponins, compounds, viz. piperine and cinnamaldehyde. The
and fats in each of the SPC samples [Table 3] were proposed methods were further validated and used for the
identified through qualitative analysis. Aqueous and quantification of these compounds. The spot at Rf 0.69 was

a b c

d e f

g h i
Figure 2: Powder microscopy of Sitopaladi churna

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Makija, et al.: Standardization of Sitopaladi churna

Table 2: Physico-chemical analysis of Sitopaladi churna samples

Parameter SPC-I* SPC-II* SPC-III*
Total ash 5.981 ± 0.103 4.639 ± 0.251 5.14 ± 0.136
Water-soluble ash 2.803 ± 0.156 2.641 ± 0.069 2.768 ± 0.117
Acid-insoluble ash 0.576 ± 0.029 0.458 ± 0.031 0.543 ± 0.021
Water-soluble extractive 35.629 ± 1.432 37.122 ± 1.467 31.17 ± 1.040
Alcohol-soluble extractive 13.026 ± 0.417 9.726 ± 0.441 11.054 ± 0.501
Loss on drying 4.826 ± 0.359 5.491 ± 0.414 4.191 ± 0.182
pH value (10% aqueous solution) 5.62 ± 0.441 5.29 ± 0.232 5.31 ± 0.130
Tap density 0.503 ± 0.025 0.546 ± 0.028 0.544 ± 0.014
Bulk density 0.481 ± 0.02 0.529 ± 0.042 0.524 ± 0.017
Angle of repose 38.387 ± 1.482 36.774 ± 1.044 36.846 ± 0.710
Haussner ratio 1.819 ± 0.01 1.166 ± 0.021 1.179 ± 0.023
Carr’s index 12.719 ± 0.475 13.402 ± 0.372 12.873 ± 0.604
*Mean (n = 3) ± SD

Table 3: Preliminary qualitative analysis of Sitopaladi churna samples

Phytochemicals SPC-I SPC-II SPC-III
Alkaloid (Dragendroff’s reagent) +ve +ve +ve
Glycoside (anisaldehyde reagent) −ve −ve −ve
Carbohydrates (anthrone reagent) +ve +ve +ve
Flavonoids (Shinoda test) +ve +ve +ve
Tannins (Folin–Denis reagent) +ve +ve +ve
Phytosterol (Libermann Burchard reagent) −ve −ve −ve
Saponins (Shaking method) +ve +ve +ve
Fats (Sudan III) +ve +ve +ve

Table 4: Total carbohydrate, tannin, and flavonoid content of Sitopaladi churna samples
Parameter SPC-I* SPC-II* SPC-III*
Aqueous extract Aqueous extract Aqueous extract
Total carbohydrate 50.293 ± 1.28 52.861 ± 1.803 49.375 ± 1.167
Total tannin content 9.297 ± 0.205 8.548 ± 0.360 8.901 ± 0.239
Total flavonoid 2.396 ± 0.137 3.774 ± 0.267 2.432 ± 0.220
content
Mean (n = 3) ± SD
*

Table 5: Powder fluorescence test of Sitopaladi churna samples

Material SPC-I SPC-II SPC-III
254 nm 366 nm 254 nm 366 nm 254 nm 366 nm
Powder as such Brown Yellow Light brown Light yellow Brown Yellow
P + 1 N NaOH Yellowish black Brown Yellowish brown Brown Brown Light brown
P + 1 N NaOH in MeOH Greenish brown Light yellow Pale green Yellow Pale green Grayish yellow
P + 1 N HCl Light green Light yellow Light green Yellow Green Pale yellow
P + 1 N H2SO4 Black Blackish brown Yellowish brown Reddish brown Brown Black
P + 1 N HNO3 Brown Dark brown Brown Reddish brown Brown Reddish brown
P + 50% KOH Yellowish brown Yellow Pale yellow Yellow Pale yellow Yellow
P + 5% FeCl3 Brown Reddish brown Brown Reddish brown Black Reddish black
P + I2 in H2O Brown Blue florescence Brown Blue florescence Black Blue florescence

identified as piperine with the help of TLC chromatograms in [Figures 4-6]. The identity of the band of piperine
of its standard using toluene:ethyl acetate:formic acid and in SPC-I, SPC-II, SPC-III extracts was confirmed by
(5:3.5:0.5 v/v/v) as the mobile phase [Figure 3]. The overlaying its UV absorption spectra with that of the
separation of piperine in SPC sample extracts is depicted standard compound [Figure 7].
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Makija, et al.: Standardization of Sitopaladi churna

The spot at Rf 0.35 was identified as cinnamaldehyde

with the help of chromatograms of its standard using
toluene:chloroform (8:2 v/v) as the mobile phase
[Figure 8]. The separation of cinnamaldehyde in SPC
sample extracts is depicted in [Figures 9-11]. The identity
of the band of cinnamaldehyde in SPC-I, SPC-II, and SPC-
III extracts was confirmed by overlaying its UV absorption
spectra with that of standard compound [Figure 12].
The purity of the band in the SPC sample extracts was
Table 6: Heavy metal estimation in Sitopaladi churna samples
Formulations Lead Mercury Arsenic Cadmium
SPC-I 8 0.24 0.53 0.27
SPC-II 7 0.14 0.24 0.16
SPC-III 11.5 0.20 0.62 0.28
Figure 3: TLC densitometric chromatogram of standard piperine at
Permissible limit as per WHO 10.0 ppm 1.0 ppm 3.0 ppm 0.3 ppm 342 nm (Rf 0.69)

Figure 4: TLC densitometric chromatogram of SPC-I at 342 nm; peak Figure 5: TLC densitometric chromatogram of SPC-II at 342 nm; peak
4: piperine (Rf 0.69) 7: piperine (Rf 0.69)

Figure 6: TLC densitometric chromatogram of SPC-III at 342 nm; peak Figure 7: Overlaying UV absorption spectra of piperine with corre-
6: piperine (Rf 0.68) sponding band in the SPC sample extracts and standards

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Makija, et al.: Standardization of Sitopaladi churna

Figure 8: TLC densitometric chromatogram of standard cinnamalde- Figure 9: TLC densitometric chromatogram of SPC-I at 310 nm; peak
hyde at 310 nm (Rf 0.35) 5: cinnamaldehyde (Rf 0.32)

Figure 10: TLC densitometric chromatogram of SPC-II at 310 nm; Figure 11: TLC densitometric chromatogram of SPC-III at 310 nm;
peak 4: cinnamaldehyde (Rf 0.32) peak 3: cinnamaldehyde (Rf 0.33)

Figure 13: Video densitometry of piperine and cinnamaldehyde. P1

and C1 are standard piperine and cinnamaldehyde, respectively; P2
Figure 12: Overlaying UV absorption spectra of cinnamaldehyde with and C2 are SPC-I samples; P3 and C3 are SPC-II samples; and P4
corresponding band in the SPC sample extracts and standards and C4 are SPC-III samples

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Makija, et al.: Standardization of Sitopaladi churna

confirmed by comparing the absorption spectra recorded cinnamaldehyde, were quantified from SPC samples by
at start, middle, and end positions of the band. The video TLC densitometric methods by HPTLC. The methods were
densitometric images of chromatoplate are depicted in validated for precision, repeatability, and accuracy. The
[Figure 13]. Preliminary TLC and co-TLC indicated the linearity ranges for piperine and cinnamaldehyde were
possible presence of piperine and cinnamaldehyde due found to be 100–1000 and 10–100 ng/spot with correlation
to the presence of P. longum and C. zeylanicumm in SPC. coefficients (r values) of 0.998 and 0.995, respectively
Hence, we quantified these two compounds from SPC [Table 7].
samples.
The TLC densitometric methods were found to be precise
TLC densitometric quantification of piperine and with %RSD for intra-day in the range of 0.98–1.18 and
cinnamaldehyde by HPTLC 1.50–1.76 and for inter-day precision in the range of 1.19–
1.44 and 1.89–2.13 for different concentrations of piperine
The two standard compounds, viz. piperine and
and cinnamaldehyde, respectively [Table 8]. This indicates
that the methods were precise and reproducible. The LOD
Table 7: Method validation parameters for the quantification of values for piperine and cinnamaldehyde were found to
piperine and cinnamaldehyde by the TLC densitometric method
be 50 and 20 ng, respectively, and LOQ values were 150
Parameter Piperine Cinnamaldehyde
and 40 ng, respectively [Table 7]. The average recoveries
Linearity range 100–1000 ng/spot 100–1000 ng/spot
at three different levels of piperine and cinnamaldehyde
Correlation coefficient 0.998 0.995
were 99.36 and 97.99%, respectively [Table 9]. Piperine
Accuracy (average % 99.36 97.99
and cinnamaldehyde were quantified from SPC-I, SPC-II,
recovery)
Instrumental precision 1.15 1.62
and SPC-III at 342 and 310 nm, respectively [Table 10]. The
(%RSD, n = 7) analytical specifications for SPC based on the above results
Repeatability (%RSD, 1.22 1.77 have been presented in Table 11.
n = 5)
LOD (ng) 50 20
LOQ (ng) 150 40 CONCLUSION
Specificity Specific Specific The present work was carried out for the formulation
and standardization of SPC. The in-house formulation
was studied for various physico-chemical parameters,
Table 8: Intra-day and inter-day precision of piperine and
cinnamaldehyde in comparison with the marketed samples. A TLC
Standard Concentration Intra-day Inter-day
densitometric method has been developed for quantification
(ng/spot) precision* precision* of piperine and cinnamaldehyde from SPC using HPTLC.
Piperine 300 1.18 1.44 The developed and validated HPTLC methods are simple,
500 1.09 1.19 precise, and accurate, and can be used for the quantification
700 0.98 1.27 of piperine and cinnamaldehyde in herbal raw materials as
Cinnamaldehyde 30 1.76 2.13 well as in their formulations. Hence, these quality-control
50 1.50 1.89 parameters and the developed HPTLC methods may be
70 1.64 1.97 considered as a tool for assistance for scientific organizations
*%RSD; mean (n = 3) and manufacturers in developing standards.

Table 9: Recovery study of piperine and cinnamaldehyde by the TLC densitometric method
Standard Amount of standard Amount of standard Amount of standard Total amount of Recovery Average
present (µg) added (%) added (µg) standard found (µg)* (%)* recovery (%)
Piperine 500 50 250 745.6 ± 0.99 99.4 ± 0.14 99.36
500 100 500 989.5 ± 2.51 98.99 ± 0.25
500 150 750 1246.47 ± 2.01 99.7 ± 0.17
Cinnamaldehyde 100 50 50 146.59 ± 1.38 97.73 ± 0.92 97.99
100 100 100 195.77 ± 0.89 97.88 ± 0.44
100 150 150 245.97 ± 1.5 98.38 ± 0.6
*Mean ± SD (n = 3)

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Table 10: Piperine and cinnamaldehyde content estimated in Pharmacognosy, Manipal College of Pharmaceutical Sciences,
Sitopaladi churna samples Manipal University, Manipal, India, for providing the funds and
Sample Piperine content Cinnamaldehyde facilities to carry out the work.
(% w/w)* (% w/w)*
SPC-I 9.51 ± 0.421 4.15 ± 0.232
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