LIMIT TESTS FOR LEAD &
ARSENIC
Limit test for Lead
Lead is the most undesirable & common impurity in drugs prepared in presence
of sulphuric acid (lead chamber process), Lead lined apparatus ,ordinary glass
bottles used for storage of chemicals and Containers opened repeatedly in
moist condition or kept without closure.
Earlier method was done using saturated Na2S solution.(Similar to H2S)
Limit test for Lead IP 96 and 2007:
Principle : Is based on the reaction between lead and dithizone (diphenyl
thiocarbazone) which results in the formation of Lead-dithizone complex in
alkaline medium.
The violet-red colored complex is compared with the standard (Pb(NO3)2, 1-10
ppm) solution.
Dithizone is soluble in chloroform and gives a green color (dithizone-chloroform
extraction solution).
The lead-dithizone complex is violet in color.
The resulting colour at the end of the process is red.
The lead present as an impurity in the substance is separated by extracting an
alkaline solution with dithizone extraction solution.
The interference by other metal ions is eliminated by adjusting the optimum pH for
the extraction by using reagents like ammonium citrate, potassium cyanide and
hydroxylamine hydrochloride
Reagents used in the Limit Test for Lead
1. Ammonium citrate, potassium cyanide, hydroxylamine hydrochloride is used to
make pH optimum so interference and influence of other impurities have been
eliminated.
2. Phenol red is used as indicator to develop the colour at the end of the
process.
3.Dithizone extraction solution: Dissolve 30mg of dithizone in 1000ml of
chloroform and add 5ml of ethanol. During use shake with 1% nitric acid and
discard the acid solution.
4.Dithizone standard solution: Dissolve 10mg of dithizone in 1000ml of
chloroform.
5.Lead nitrate standard solution ( 1ppm):Dissolve 1 volume of lead standard
solution (10ppm Pb) to 10 volumes with water.
6. Nitric acid solution (1%)
7. Sample solution containing lead present as an impurities gets separated by
extracting an alkaline solution with a dithizone extraction solution.
Procedure :
Observation :
The intensity of the red colour complex is depending on the amount of lead in the
solution.
The colour produced in the sample solution should not be greater than standard
solution.
If the colour produced in sample or test solution is less than the standard
solution, the sample will pass the limit test of the lead and vice versa.
A few typical examples of Pb+ impurity from the official compendium are given
below :
Arsenic risk
Arsenic Limit Test:
Earth’s crust is an abundant natural source of arsenic in the form of
“Arsenopyrite”.
Inorganic arsenic is found in ground water used for drinking.
Arsenic produces cumulative toxicity by Contaminated water used for drinking,
food preparation and irrigation of food crops poses the greatest threat to public
health from arsenic. Long-term exposure to arsenic from drinking-water and food
can cause cancer and skin lesions.
Principle : The test is based on the reduction of arsenic in arsenious state to
arsine gas (arsenious hydride).
Arsine gas on passing over mercuric chloride paper produces mercuric arsenide
yellow to brown stain.
Mercuric chloride paper
Mercuric arsenide stain
Principle:
The sample when dissolved in acidic media, converts arsenic impurity into arsenious
acid and arsenic acid depending upon the valency state.
OH
+
As+3 H H3 ASO3 As OH Arsenious acid
Trivalent OH
arsenic
OH
As+5 H+ H3 ASO4 As OH Arsenic acid
O
Pentavalent
arsenic OH
In presence of KI and stannated HCl as reducing agents, arsenic acid is converted into
arsenious acid.
Arsenious acid on reaction with nascent hydrogen (produced by Zn/HCl) to produce arsenious
gas or arsenious hydride.
Zn+2HCl ZnCl2+2[H]
H3AsO3 + 3H2 AsH3 + 3H2O
Arsine gas/
Arsenious hydride
Arsine gas produces yellow to brown stain on dry mercuric chloride paper due to the
formation of mercuric arsenide complex.
2 AsH3 + HgCl2 Hg (AsH2)2 + 2HCl
AsH2
Hg
AsH2
Mercuric arsenide
Yellow to brown stain
Lead acetate cotton traps H2S. (Commercial Zn granules contain sulphides, sulphates and
thiosulphates as impurities and when these impurities react with nascent hydrogen forms H2S.
This if not trapped, forms a black colored stain on the HgCl2 paper and hinder the actual stain
(CH3COO)2Pb + H2S PbS + CH3COOH
Lead
Lead acetate sulphide
Zn granules only are used and not the powder, why?.
Zn powder if used there will be a vigorous evolution of hydrogen for a short period of
time and so the complete reduction is doubted.
To obtain nascent hydrogen in steady and longer duration IP prescribes granular Zinc.
Reagents used:
All special reagents used in the test for arsenic are marked arsenic free as AsT
and used in LT of Arsenic only.
1. Stannous Chloride or Stannated Hydrochloric acid AsT : It is prepared from
solution of Stannous Chloride by adding an equal volume of hydrochloric acid ,
boiling down to original volume and filtering through the fine grind filter
paper . This is used for complete evolution of arsine gas as well as reducing
agent to reduce arsenic to arsenious and then to arsine.
2. Zinc grannules, Potassium Iodide,
3. Lead acetate soaked cotton plug is used to trap any hydrogen sulphide
which may be evolved along with arsine.
Gutzeit’s Apparatus description
Consists of a wide mouthed glass bottle of 120ml capacity having a mouth of
about 2.5cm in diameter.
This is fitted with a rubber bung through which passes a glass tube having a
external diameter of 0.8cm and internal diameter of 0.65cm.
The tube is constricted at its lower and extremity to about 1mm diameter and
there is a hole not less than 2mm diameter in the side of the tube near the
constricted part.
The upper end of the glass tube has been fitted with two rubber bungs (
about 25mmx25mm) each having a hole bored inside centrally and exactly
6.5mm in diameter.
One of the bungs is fitted to the upper end of the tube while the second bung
has to be fitted upon the first bung in such a way that the mercuric chloride
paper gets exactly sandwiched between the central perforation of the two
The bungs are kept close contact by using rubber band or clip in such a
manner that gas evolved from the bottle must have to pass through the
0.65cm internal circle of the mercuric chloride paper.
Procedure:
Sample is dissolved in water as per mentioned in individual monograph.
Transfer into Gutzeit’s apparatus.
Add 1 gm of Potassium Iodide (5ml 0f 1M KI), 5ml of Stannated HCl solution
and 10gm of Zn grannules AsT.
Close the bottle with set upper assembly and allow the reaction for 40min to
1 hr. to take place. Most suitable reaction temperature should be 400C.
Dismantle the assembly and observe the mercuric chloride paper.
Compare it with the standard stain produced by treating 1.0ml of Arsenic
standard solution (10ppm) diluted to 50ml with water in the same way as
sample.
Observation: If the intensity of the test yellow stain is less than the standard
the sample passes or complies with the LT for Arsenic.
The colour of the stain fades quickly so has to be compared immediately. It
can be preserved by applying wax.
