A212 Appendix VII

desired, as the specified range is approached), dilute with

water to 40 ml and mix.
Add 2 ml of buffer solution pH 3.5 and 1.2 ml of thioacetamide
reagent. Mix immediately. Dilute to 50 ml with water and
mix.
REFERENCE SOLUTION

Prepare at the same time and in the same manner using the
prescribed volume of lead standard solution (10 ppm Pb).
After 2 min, any brown colour in the test solution is not
more intense than that in the reference solution.

Limit Test for Iron

(Ph. Eur. method 2.4.9)
Dissolve the specified quantity of the substance being
examined in sufficient water to produce 10 ml or use 10 ml
of the solution prescribed in the monograph and transfer to a
Nessler cylinder. Add 2 ml of a 20% w/v solution of citric acid
and 0.1 ml of ,nercaptoacetic acid, mix, make alkaline with
10m armonio, dilute to 20 ml with water and allow to stand
for 5 minutes. Any pink colour produced is not more intense
than that obtained by treating 10 ml of iron standard solution
(1 ppm Fe) in the same manner.

Limit Test for Lead in Sugars

(Ph. Eur. method 2.4.10)
Carry out Method II for atomic absorption spectrophotornetry,
Appendix II D, using an airacetylene flame, a lead
hollow-cathode lamp and the following solutions.
TEST SOLUTION

Dissolve 20.0 g of the substance being examined in sufficient

lvi acetic acid to produce 100.0 ml. Add 2.0 ml of a clear
1% w/v solution of ammonium pyrrolidinedithiocarbamate and
10.0 ml of 4-methylpentan-2-one and shake for 30 seconds
protected from bright light.
REFERENCE SOLUTIONS

Prepare three reference solutions in the same manner as the

test solution using 0.5 ml, 1.0 ml and 1.5 ml respectively of
lead standard solution (10 ppm Pb) in addition to 20.0 g of the
substance being examined.
Prepare a blank solution in the same manner as the test
solution but without the substance being examined.
Examine the test solution, the three reference solutions and
the blank solution at the maximum at 283.3 nm using the
blank solution to set the zero of the instrument. Construct a
calibration curve and determine the content of lead in the
substance being examined. The lead content is not greater
than 0.5 ppm, unless otherwise prescribed.

Limit Test for Magnesium

(Ph. Eur. method 2.4.6)
To 10 ml of a solution prepared as specified in the
monograph add 0.1 g of sodium tetraborate. Adjust the pH of
the solution, if necessary, to 8.8 to 9.2 with 2m hydrochloric
acid or 2m sodium hydroxide. Shake with two 5-ml quantities
of a 0.1% w/v solution of 8-hydroxyquinoline in chloroform,
shaking for 1 minute, allowing to stand, separating and
discarding the organic layer each time. To the aqueous layer
add 0.4 ml of n-butylamine and 0.1 ml of triethanolamine.
Adjust the pH of the solution, if necessary, to 10.5 to 11.5.
Add 4 ml of the solution of 8-hydroxyquinoline, shake for
1 minute, allow to stand and separate. Any colour produced
in the lower layer is not more intense than that obtained by
treating a mixture of 1 ml of magnesiurn standard solution
(10 ppm mg) and 9 ml of water in the same manner.

Limit Test for Magnesium and Alkaline-earth

Metals
(Ph. Eur. method 2.4.7)
To 200 ml of water add 0.1 g of hydroxylamine hydrochloride,
10 ml of ammonia buffer pH 10.0, 1 ml of 0.1M zinc sulphate
and about 15 mg of mordant block 11 triturate. Heat to about
40' and titrate with 0.01m disodium edetate VS until the violet
colour changes to a full blue. To the solution add the
specified quantity of the substance being examined dissolved
in 100 ml of water or use the prescribed solution. If the
colour of the solution changes to violet, titrate with
0.01m disodium edetate VS until the full blue colour is again
produced. The volume of 0.01m disodium edetate VS used in
the second titration does not exceed the prescribed quantity.

Heavy Metals in Herbal Drugs and Fatty Oils

(Ph. Eur. method 2.4.27)
Examine by atomic absorption spectrometry (2.2.23).
Caution: when using closed high-pressure digestion vessels
and microwave laboratory equipment, be familiar with the
safety and operating instructions given by the manufacturer.

APPARATUS
The apparatus typically consists of the following:
as digestion flasks, polytetrafluoroethylene flasks with a
volume of about 120 ml, fitted with an airtight closure, a
valve to adjust the pressure inside the container and a
polytetrafluoroethylene tube to allow release of gas,
a system to make flasks airtight, using the same torsional
force for each of them,
a microwave oven, with a magnetron frequency of
2450 MHz, with a selectable output from O to 630 70 W
in 1 per cent increments, a programmable digital
computer, a polytetrafluoroethylene-coated microwave
cavity with a variable speed exhaust fan, a rotating
turntable drive system and exhaust tubing to vent fumes,
an atomic absorption spectrometer, equipped with hollowcathode lamps as source of radiation and a deuterium
lamp as background corrector; the system is fitted with:
(a) a graphite furnace as atomisation device for cadmium,
copper, iron, lead, nickel and zinc.
(b) an automated continuous-flow hydride vapour
generation system for arsenic and mercury.

METHOD
In case alternative apparatus is used, an adjustment of the
instrument parameters may be necessary.
Clean all the glassware and laboratory equipment with a
10 g/I solution of nitric acid R before use.
Test solution
In a digestion flask place the prescribed quantity of the
substance to be examined (about 0.50 g of powdered drug
(1400) or 0.50 g of fatty oil). Add 6 ml of heavy metal-free
nitric acid R and 4 ml of heavy metal-free hydrochloric
acid R. Make the flask airtight.
Place the digestion flasks in the microwave oven. Carry out
the digestion in 3 steps according to the following
programme, used for 7 flasks each containing the test
solution: 80 per cent power for 15 min, 100 per cent power
for 5 min, 80 per cent power for 20 min.
At the end of the cycle allow the flasks to cool in air and to
each add 4 ml of heavy metal-free sulphuric acid R. Repeat
the digestion programme. After cooling in air, open each
digestion flask and introduce the clear, colourless solution