518 Benserazide Hydrochloride / Official Monographs JP XVIII

System repeatability: When the test is repeated 6 times

with 50 mL of the standard solution under the above operat- Benserazide Hydrochloride
ing conditions, the relative standard deviation of the peak
area of benidipine is not more than 1.5z. ベンセラジド塩酸塩
Assay Weigh accurately the mass of not less than 20
Benidipine Hydrochloride Tablets, and powder using an
agate mortar. Weigh accurately a part of the powder,
equivalent to about 8 mg of benidipine hydrochloride
(C28H31N3O6.HCl), add about 150 mL of a mixture of
diluted phosphoric acid (1 in 500) and methanol (1:1), shake, C10H15N3O5.HCl: 293.70
then add the mixture of diluted phosphoric acid (1 in 500) (2RS )-2-Amino-3-hydroxy-
and methanol (1:1) to make exactly 200 mL, and centrifuge. N?-(2,3,4-trihydroxybenzyl)propanoylhydrazide
Pipet 20 mL of the supernatant liquid, add exactly 10 mL of monohydrochloride
the internal standard solution and the mixture of diluted [14919-77-8]
phosphoric acid (1 in 500) and methanol (1:1) to make 50
mL, and use this solution as the sample solution. Separately, Benserazide Hydrochloride contains not less than
weigh accurately about 40 mg of benidipine hydrochloride 98.0z and not more than 101.0z of benserazide hy-
for assay, previously dried at 1059 C for 2 hours, and dis- drochloride (C10H15N3O5.HCl), calculated on the an-
solve in the mixture of diluted phosphoric acid (1 in 500) and hydrous basis.
methanol (1:1) to make exactly 100 mL. Pipet 2 mL of this
Description Benserazide Hydrochloride occurs as a white
solution, add exactly 10 mL of the internal standard solution
to grayish white crystalline powder.
and the mixture of diluted phosphoric acid (1 in 500) and
It is freely soluble in water and in formic acid, soluble in
methanol (1:1) to make 50 mL, and use this solution as the
methanol, very slightly soluble in ethanol (95).
standard solution. Perform the test with 10 mL each of the
It dissolves in 0.1 mol/L hydrochloric acid TS.
sample solution and standard solution as directed under Liq-
The pH of a solution of 1.0 g of Benserazide Hydrochlo-
uid Chromatography <2.01> according to the following con-
ride in 100 mL of water is between 4.0 and 5.0.
ditions, and calculate the ratios, QT and QS, of the peak area
It is hygroscopic.
of benidipine to that of the internal standard.
It is gradually colored by light.
Amount (mg) of benidipine hydrochloride A solution of Benserazide Hydrochloride (1 in 100) shows
(C28H31N3O6.HCl) no optical rotation.
＝ MS × QT/QS × 1/5
Identification (1) Determine the absorption spectrum of a
MS: Amount (mg) of benidipine hydrochloride for assay solution of Benserazide Hydrochloride in 0.1 mol/L hydro-
taken chloric acid TS (1 in 10,000) as directed under Ultraviolet-
visible Spectrophotometry <2.24>, and compare the spectrum
Internal standard solution—A solution of benzoin in a mix-
with the Reference Spectrum: both spectra exhibit similar in-
ture of water and methanol (1:1) (13 in 200,000).
tensities of absorption at the same wavelengths.
Operating conditions—
(2) Determine the infrared absorption spectrum of Ben-
Detector: An ultraviolet absorption photometer (wave-
serazide Hydrochloride as directed in the potassium chloride
length: 237 nm).
disk method under Infrared Spectrophotometry <2.25>, and
Column: A stainless steel column 4.6 mm in inside diame-
compare the spectrum with the Reference Spectrum: both
ter and 10 cm in length, packed with octadecylsilanized silica
spectra exhibit similar intensities of absorption at the same
gel for liquid chromatography (3 mm in particle diameter).
wave numbers.
Column temperature: A constant temperature of about
(3) To 10 mL of a solution of Benserazide Hydrochloride
259 C.
(1 in 30) add silver nitrate TS: a white precipitate is formed.
Mobile phase: A mixture of 0.05 mol/L potassium dihy-
To a portion of this precipitate add dilute nitric acid: the
drogen phosphate TS (pH 3.0), methanol and tetrahydrofu-
precipitation does not dissolve.
ran (65:27:8).
Flow rate: Adjust so that the retention time of benidipine Purity (1) Clarity and color of solution—Dissolve 0.5 g
is about 20 minutes. of Benserazide Hydrochloride in 10 mL of water, and per-
System suitability— form the test with this solution as directed under Ultraviolet-
System performance: When the procedure is run with 10 visible Spectrophotometry <2.24>: the absorbance of this so-
mL of the standard solution under the above operating con- lution at 430 nm is not more than 0.10.
ditions, the internal standard and benidipine are eluted in (2) Heavy metals <1.07>—Proceed with 1.0 g of Benser-
this order with the resolution between these peaks being not azide Hydrochloride according to Method 2, and perform
less than 8. the test. Prepare the control solution with 2.0 mL of Stand-
System repeatability: When the test is repeated 6 times ard Lead Solution (not more than 20 ppm).
with 10 mL of the standard solution under the above operat- (3) Related substances—Conduct this procedure without
ing conditions, the relative standard deviation of the ratio of exposure to light, using light-resistant vessels. Dissolve 0.25
the peak area of benidipine to that of the internal standard is g of Benserazide Hydrochloride in 10 mL of methanol, and
not more than 1.0z. use this solution as the sample solution. Pipet 1 mL and 3
mL of the sample solution, add methanol to make exactly
Containers and storage Containers—Well-closed contain-
200 mL, and use these solutions as the standard solution (1)
ers.
and (2), respectively. Perform the test with these solutions as
directed under Thin-layer Chromatography <2.03>. Spot 2
mL each of the sample solution and standard solutions (1)
and (2) on a plate of cellulose for thin-layer chromatogra-

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.)
JP XVIII Official Monographs / Benzalkonium Chloride 519

phy. Develop the plate with a solution of formic acid in so- as follows: mix 2.5 mL of Standard Lead Solution, 0.15 g of
dium chloride TS (1 in 1000) to a distance of about 10 cm, hydroxylammonium chloride, 0.15 g of sodium acetate trihy-
and air-dry the plate. Spray evenly sodium carbonate TS, drate, and 2 mL of dilute acetic acid, and add water to make
air-dry, and then spray evenly Folin's TS on the plate: the 50 mL (not more than 50 ppm).
spots other than the principal spot obtained from the sample (2) Arsenic <1.11>—To 1.0 g of Bentonite add 5 mL of
solution are not more intense than the spot from the stand- dilute hydrochloric acid, and gently heat to boil while stir-
ard solution (2), and the number of the spots which intense ring well. Cool immediately, and centrifuge. To the residue
more than the spot from the standard solution (1) are not add 5 mL of dilute hydrochloric acid, shake well, and centri-
more than 2. fuge. To the residue add 10 mL of water, and perform the
same operations. Combine all the extracts, and heat on a
Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-
water bath to concentrate to 5 mL. Perform the test with this
tion, direct titration). Use a solution of salicylic acid in
solution as the test solution (not more than 2 ppm).
methanol for water determination (3 in 20) instead of metha-
(3) Foreign matter—Place 2.0 g of Bentonite in a mor-
nol for water determination.
tar, add 20 mL of water to swell, disperse evenly with a pes-
Residue on ignition <2.44> Not more than 0.1z (1 g). tle, and dilute with water to 100 mL. Pour the suspension
through a No. 200 (74 mm) sieve, and wash the sieve thor-
Assay Weigh accurately about 0.3 g of Benserazide Hydro-
oughly with water. No grit is felt when the fingers are rubbed
chloride, dissolve in 5 mL of formic acid, add 50 mL of ace-
over the wire mesh of the sieve.
tic acid (100), and titrate <2.50> immediately with 0.1 mol/L
perchloric acid VS (potentiometric titration). Perform a Loss on drying <2.41> 5.0 – 10.0z (2 g, 1059C, 2 hours).
blank determination in the same manner, and make any nec-
Gel formation Mix 6.0 g of Bentonite with 0.30 g of mag-
essary correction.
nesium oxide. Add the mixture, in several portions, to 200
Each mL of 0.1 mol/L perchloric acid VS mL of water contained in a glass-stoppered 500-mL cylinder.
＝ 29.37 mg of C10H15N3O5.HCl Agitate for 1 hour, transfer 100 mL of the suspension to a
100-mL graduated cylinder, and allow to stand for 24 hours:
Containers and storage Containers—Tight containers.
not more than 2 mL of supernatant appears on the surface.
Storage—Light-resistant.
Swelling power To 100 mL of water in a glass-stoppered
100-mL cylinder add 2.0 g of Bentonite in ten portions,
Bentonite allowing each portion to settle before adding the next, and
allow to stand for 24 hours: the apparent volume of the sedi-
ベントナイト ment at the bottom is not less than 20 mL.
Containers and storage Containers—Well-closed contain-
Bentonite is a natural, colloidal, hydrated aluminum ers.
silicate.
Description Bentonite occurs as a very fine, white to light
yellow-brown powder. It is odorless. It has a slightly earthy Benzalkonium Chloride
taste.
ベンザルコニウム塩化物
It is practically insoluble in water, in ethanol (95) and in
diethyl ether.
It swells in water. Benzalkonium Chloride is represented by the for
mula [C6H5CH2N(CH3)2R]Cl, in which R extends
Identification (1) Add 0.5 g of Bentonite to 3 mL of
from C8H17 to C18H37, with C12H25 and C14H29 com-
diluted sulfuric acid (1 in 3), and heat until white fumes are
prising the major portion.
evolved. Cool, add 20 mL of water, and filter. To 5 mL of
It contains not less than 95.0z and not more than
the filtrate add 3 mL of ammonia TS: a white, gelatinous
105.0z of benzalkonium chloride (as C22H40ClN:
precipitate is produced, which turns red on the addition of 5
354.01), calculated on the anhydrous basis.
drops of alizarin red S TS.
(2) Wash the residue obtained in (1) with water, add 2 Description Benzalkonium Chloride occurs as a white to
mL of a solution of methylene blue trihydrate (1 in 10,000), yellow-white powder, colorless to light yellow, gelatinous
and wash again with water: the residue is blue in color. pieces, or jelly-like fluid or mass. It has a characteristic
odor.
pH <2.54> To 1.0 g of Bentonite add 50 mL of water, and
It is very soluble in water and in ethanol (95), and practi-
shake: the pH of the suspension is between 9.0 and 10.5.
cally insoluble in diethyl ether.
Purity (1) Heavy metals <1.07>—To 1.5 g of Bentonite A solution of Benzalkonium Chloride foams strongly
add 80 mL of water and 5 mL of hydrochloric acid, and boil when shaken.
gently for 20 minutes with thorough stirring. Cool, centri-
Identification (1) Dissolve 0.2 g of Benzalkonium Chlo-
fuge, collect the supernatant liquid, wash the residue with
ride in 1 mL of sulfuric acid, add 0.1 g of sodium nitrate,
two 10-mL portions of water, and centrifuge each. Combine
and heat for 5 minutes on a water bath. After cooling, add
the supernatant liquid and the washings, and add dropwise
10 mL of water and 0.5 g of zinc powder, heat for 5 minutes,
ammonia solution (28). When a precipitate is produced, add
cool, and filter: the filtrate responds to Qualitative Tests
dropwise dilute hydrochloric acid with vigorous stirring, and
<1.09> for primary aromatic amines. The color of the solu-
dissolve. To the solution add 0.45 g of hydroxylammonium
tion is red.
chloride, and heat. Cool, and add 0.45 g of sodium acetate
(2) To 2 mL of a solution of Benzalkonium Chloride (1
trihydrate, 6 mL of dilute acetic acid and water to make 150
in 1000) add a mixture of 0.2 mL of a solution of bromophe-
mL. Pipet 50 mL of the solution, and perform the test using
nol blue (1 in 2000) and 0.5 mL of sodium hydroxide TS: a
this solution as the test solution. Prepare the control solution

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.)