JP XVIII General Tests / 1.

07 Heavy Metals Limit Test 29

potentiometric titration. Perform the test with the blank so- and designate it as the test solution.
lution in the same manner, and make any necessary correc- The control solution is prepared by placing the volume of
tion. Standard Lead Solution directed in the monograph in a
Nessler tube, and adding 2 mL of dilute acetic acid and
Each mL of 0.005 mol/L silver nitrate VS
water to make 50 mL.
＝ 0.6345 mg of I
1.2. Method 2
3.3. Fluorine Place an amount of the sample, directed in the mono-
Apply a small amount of water to the upper part of A, graph, in a quartz or porcelain crucible, cover loosely with a
pull out C carefully, transfer the test solution and the blank lid, and carbonize by gentle ignition. After cooling, add 2
solution to 50 mL volumetric flasks separately, wash C, B mL of nitric acid and 5 drops of sulfuric acid, heat cau-
and the inner side of A with water, add the washings and tiously until white fumes are no longer evolved, and inciner-
water to make 50 mL, and use these solutions as the test so- ate by ignition between 5009 C and 6009C. Cool, add 2 mL
lution and the correction solution. Pipet the test solution of hydrochloric acid, evaporate to dryness on a water bath,
(V mL) equivalent to about 30 mg of fluorine, V mL of the moisten the residue with 3 drops of hydrochloric acid, add
correction solution and 5 mL of standard fluorine solution, 10 mL of hot water, and warm for 2 minutes. Then add 1
transfer to 50-mL volumetric flasks separately, add 30 mL of drop of phenolphthalein TS, add ammonia TS dropwise
a mixture of alizarin complexone TS, acetic acid-potassium until the solution develops a pale red color, add 2 mL of
acetate buffer solution (pH 4.3) and cerium (III) nitrate TS dilute acetic acid, filter if necessary, and wash with 10 mL of
(1:1:1), add water to make 50 mL, and allow to stand for 1 water. Transfer the filtrate and washings to a Nessler tube,
hour. Perform the test with these solutions as directed under and add water to make 50 mL. Designate it as the test solu-
Ultraviolet-visible Spectrophotometry <2.24>, using a blank tion.
prepared with 5 mL of water in the same manner. Determine The control solution is prepared as follows: Evaporate a
the absorbances, AT, AC and AS, of the subsequent solutions mixture of 2 mL of nitric acid, 5 drops of sulfuric acid and 2
of the test solution, the correction solution and the standard mL of hydrochloric acid on a water bath, further evaporate
solution at 600 nm. to dryness on a sand bath, and moisten the residue with 3
drops of hydrochloric acid. Hereinafter, proceed as directed
Amount (mg) of fluorine (F) in the test solution
in the test solution, then add the volume of Standard Lead
＝ amount (mg) of fluorine in 5 mL of
Solution directed in the monograph and water to make 50
AT － A C 50 mL.
the standard solution × ×
AS V 1.3. Method 3
Place an amount of the sample, directed in the mono-
Standard Fluorine Solution: Dry sodium fluoride (stand-
graph, in a quartz or porcelain crucible, heat cautiously,
ard reagent) in a platinum crucible between 5009C and
gently at first, and then incinerate by ignition between 5009 C
5509C for 1 hour, cool it in a desiccator (silica gel), weigh ac-
and 6009 C. After cooling, add 1 mL of aqua regia, evapo-
curately about 66.3 mg of it, and dissolve in water to make
rate to dryness on a water bath, moisten the residue with 3
exactly 500 mL. Pipet 10 mL of this solution, and dilute with
drops of hydrochloric acid, add 10 mL of hot water, and
sufficient water to make exactly 100 mL.
warm for 2 minutes. Add 1 drop of phenolphthalein TS, add
3.4. Sulfur
ammonia TS dropwise until the solution develops a pale red
Apply a small amount of water to the upper part of A,
color, add 2 mL of dilute acetic acid, filter if necessary, wash
pull out C carefully, and wash C, B and the inner side of A
with 10 mL of water, transfer the filtrate and washings to a
with 15 mL of methanol. To this solution add 40 mL of
Nessler tube, and add water to make 50 mL. Designate it as
methanol, then add exactly 25 mL of 0.005 mol/L barium
the test solution.
perchlorate VS, allow to stand for 10 minutes, add 0.15 mL
The control solution is prepared as follows: Evaporate 1
of arsenazo III TS with a measuring pipet, and titrate <2.50>
mL of aqua regia to dryness on a water bath. Hereinafter,
with 0.005 mol/L sulfuric acid VS. Perform the test with the
proceed as directed for the test solution, and add the volume
blank solution in the same manner.
of Standard Lead Solution directed in the monograph and
Each mL of 0.005 mol/L barium perchlorate VS water to make 50 mL.
＝ 0.1604 mg of S 1.4. Method 4
Place an amount of the sample, directed in the mono-
graph, in a platinum or porcelain crucible, mix with 10 mL
of a solution of magnesium nitrate hexahydrate in ethanol
1.07 Heavy Metals Limit Test (95) (1 in 10), fire the ethanol to burn, and carbonize by
gradual heating. Cool, add 1 mL of sulfuric acid, heat care-
Heavy Metals Limit Test is a limit test of the quantity of
fully, and incinerate by ignition between 5009 C and 6009C.
heavy metals contained as impurities in drugs. The heavy
If a carbonized substance remains, moisten with a small
metals are the metallic inclusions that are darkened with so-
amount of sulfuric acid, and incinerate by ignition. Cool,
dium sulfide TS in acidic solution, as their quantity is ex-
dissolve the residue in 3 mL of hydrochloric acid, evaporate
pressed in terms of the quantity of lead (Pb).
on a water bath to dryness, wet the residue with 3 drops of
In each monograph, the permissible limit for heavy metals
hydrochloric acid, add 10 mL of water, and dissolve by
(as Pb) is described in terms of ppm in parentheses.
warming. Add 1 drop of phenolphthalein TS, add ammonia
1. Preparation of test solutions and control solutions TS dropwise until a pale red color develops, then add 2 mL
Unless otherwise specified, test solutions and control solu- of dilute acetic acid, filter if necessary, wash with 10 mL of
tions are prepared as directed in the following: water, transfer the filtrate and the washing to a Nessler tube,
1.1. Method 1 add water to make 50 mL, and use this solution as the test
Place an amount of the sample, directed in the mono- solution.
graph, in a Nessler tube. Dissolve in water to make 40 mL. The control solution is prepared as follows: Take 10 mL
Add 2 mL of dilute acetic acid and water to make 50 mL, of a solution of magnesium nitrate hexahydrate in ethanol

The JP Drugs are to be tested according to the provisions given in the pertinent monographs, General Notices, General Rules for Crude Drugs,
General Rules for Preparations, and General Tests for their conformity to the Japanese Pharmacopoeia. (See the General Notices 5.)
30 1.08 Nitrogen Determination (Semimicro-Kjeldahl Method) / General Tests JP XVIII
(95) (1 in 10), and fire the ethanol to burn. Cool, add 1 mL
of sulfuric acid, heat carefully, and ignite between 5009C
and 6009C. Cool, and add 3 mL of hydrochloric acid. Here-
inafter, proceed as directed in the test solution, then add the
volume of Standard Lead Solution directed in the mono-
graph and water to make 50 mL.
2. Procedure
Add 1 drop of sodium sulfide TS to each of the test solu-
tion and the control solution, mix thoroughly, and allow to
stand for 5 minutes. Then compare the colors of both solu-
tions by viewing the tubes downward or transversely against
a white background. The test solution has no more color
than the control solution.

1.08 Nitrogen Determination

(Semimicro-Kjeldahl Method)
Nitrogen Determination is a method to determine nitrogen
in an organic substance in which the nitrogen is converted
into ammonia nitrogen by thermal decomposition of the or-
ganic substance with sulfuric acid, and the ammonia liber-
ated by alkali and trapped by distillation with steam is deter-
mined by titration.
1. Apparatus
Use the apparatus illustrated in Fig. 1.08-1. It is thor-
oughly constructed of hard glass, and ground glass surfaces
may be used for joints. All rubber parts used in the appa-
ratus should be boiled for 10 to 30 minutes in sodium hy-
droxide TS and for 30 to 60 minutes in water, and finally
washed thoroughly with water before use.
Alternatively, apparatus can be used in which some of the
procedures, such as digestion of organic substances, distilla-
tion of the liberated ammonia, and endpoint detection
methods in titrimetry (e.g., potentiometric titration or titra-
tion by colorimeter) are automated. Fig. 1.08-1

2. System suitability
If an automated apparatus is used, it is necessary to con-
firm periodically the suitability of the apparatus according to
Then, while shaking the flask, add cautiously 1 mL of
the following method:
hydrogen peroxide (30) drop by drop along the inside wall of
Weigh accurately about 1.7 g of amidosulfuric acid (stand-
the flask. Heat the flask gradually, then heat so strong that
ard reagent), previously dried in a desiccator (in vacuum,
the vapor of sulfuric acid is condensed at the neck of the
silica gel) for about 48 hours, dissolve in water to make ex-
flask, until the solution changes through a blue and clear to
actly 200 mL. Pipet 2 mL of this solution, and transfer to a
a vivid green and clear, and the inside wall of the flask is free
digestion flask. When the test is performed as directed in the
from a carbonaceous material. If necessary, add a small
instrumental manual the nitrogen content (z) in amidosul-
quantity of hydrogen peroxide (30) after cooling, and heat
furic acid should be determined between 14.2z and 14.6z.
again. After cooling, add cautiously 20 mL of water, cool
3. Reagents, Test Solutions the solution, and connect the flask to the distillation appa-
Decomposition accelerator: Unless otherwise specified, ratus (Fig. 1.08-1) washed beforehand by passing steam
use 1 g of a powdered mixture of 10 g of potassium sulfate through it. To the absorption flask K add 15 mL of boric
and 1 g of cupper (II) sulfate pentahydrate. The composition acid solution (1 in 25), 3 drops of bromocresol green-methyl
and amount of the digestion accelerator may be modified if red TS and sufficient water to immerse the lower end of the
it is confirmed that the modified one give almost the same condenser tube J. Add 30 mL of sodium hydroxide solution
results using the sample as those obtained from the conven- (2 in 5) through the funnel F, rinse cautiously the funnel with
tional catalyst. 10 mL of water, close the clamp attached to the rubber tub-
ing G, then begin the distillation with stream, and continue
4. Procedure
until the distillate measures 80 to 100 mL. Remove the ab-
Unless otherwise specified, proceed by the following
sorption flask from the lower end of the condenser tube J,
method. Weigh accurately or pipet a quantity of the sample
rinsing the end part with a small quantity of water, and
corresponding to 2 to 3 mg of nitrogen (N:14.01), and place
titrate <2.50> the distillate with 0.005 mol/L sulfuric acid VS
in the Kjeldahl flask A. Add the decomposition accelerator
until the color of the solution changes from green through
and wash down any adhering sample from the neck of the
pale grayish blue to pale grayish red-purple. Perform a blank
flask with a small quantity of water. Add 7 mL of sulfuric
determination in the same manner, and make any necessary
acid, allowing it to flow down the inside wall of the flask.
correction.

The JP Drugs are to be tested according to the provisions given in the pertinent monographs, General Notices, General Rules for Crude Drugs,
General Rules for Preparations, and General Tests for their conformity to the Japanese Pharmacopoeia. (See the General Notices 5.)