Supplement I to the Japanese Pharmacopoeia Fourteenth Edition

Supplement I, JP XIV
O‹cial Monographs for Part I
1511
Amount [ mg (potency)] of pivmecillinam (C 21 H 33 N 3 O 5 S)
= W S × Q T
Q S
× 1000
W S : Amount [mg (potency)] of Pivmecillinam Hydrochloride
Reference Standard
Internal standard solution—A solution of diphenyl in the
mobile phase (1 in 12,500).
Operating conditions—
Detector: An ultraviolet absorption photometer
(wavelength: 254 nm).
Column: A stainless steel column 4 mm in inside diameter
and 30 cm in length, packed with octadecylsilanized silica gel
for liquid chromatography (10 mm in particle diameter).
Column temperature: A constant temperature of about
259C.
Mobile phase: Dissolve 0.771 g of ammonium acetate in
about 900 mL of water, adjust the pH to 3.5 with acetic acid
(100), and add water to make 1000 mL. To 400 mL of this
solution add 600 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention time
of pivmecillinam is about 6.5 minutes.
System suitability—
System performance: When the procedure is run with 10
mL of the standard solution under the above operating conditions,
pivmecillinam and the internal standard are eluted
in this order with the resolution between these peaks being
not less than 4.
System repeatability: When the test is repeated 6 times
with 10 mL of the standard solution under the above operating
conditions, the relative standard deviation of the ratios
of the peak area of pivmecillinam to that of the internal
standard is not more than 1.0z.
Identiˆcation (1) To 5 mL of a solution of Polymixin B
Sulfate (1 in 10) add 5 mL of a solution of sodium hydroxide
(1 in 10), add 5 drops of a solution of copper (II) sulfate pentahydrate
(1 in 100) while shaking: a purple color develops.
(2) Transfer 5 mg each of Polymixin B Sulfate and Polymixin
B Sulfate Reference Standard separately in two glass
stoppered test tubes, add 1 mL of diluted hydrochloric acid
(1 in 2), stopper the tube, heat at 1359C for 5 hours, then
heat to dryness on a water bath, and keep the heating until
no more hydrochloric acid odor is evolved. Dissolve the
residue in 0.5 mL of water, and use these solutions as the
sample solution and the standard solution (1). Separately,
dissolve 20 mg each of L-leucine, L-threonine, phenylalanine
and L-serine separately in 10 mL of water, and use these solutions
as the standard solution (2), the standard solution
(3), the standard solution (4) and the standard solution (5),
respectively. Perform the test with these solutions as directed
under the Thin-layer Chromatography. Spot 3 mL eachof
the sample solution, the standard solution (1), the standard
solution (2), the standard solution (3), the standard solution
(4) and the standard solution (5) on a plate of silica gel for
thin-layer chromatography, and expose the plate to a saturated
vapor of the developing solvent for 15 hours. Develop
the plate with a mixture of phenol and water (3:1) to a distance
of about 13 cm while without exposure to light, and
dry the plate at 1109C for 5 minutes. Spray evenly ninhydrin-acetic
acid TS on the plate, and heat at 1109C for5
minutes: Rf value of each spot obtained from the sample solution
is the same with Rf value of the corresponding spots
from the standard solution (1). Each of the spots from the
sample solution appears at the position corresponding to
each of the spots from the standard (2), (3) and (4), but not
appears at the position corresponding to the spot from the
standard solution (5).
(3) A solution of Polymixin B Sulfate (1 in 20) responds
to the Qualitative Tests for sulfate.
Containers and storage
Containers—Tight containers.
Optical rotation [a] 20 D : -78 – -909(0.5 g calculated on
the dried basis, water, 25 mL, 100 mm).
Polymixin B Sulfate
硫 酸 ポリミキシン B
Change to read except the structural formula
and chemical name:
Polymixin B Sulfate contains not less than 6500
units per mg, calculated on the dried basis. The potency
of Polymixin B Sulfate is expressed as mass unit of
polymixin B (C 55-56 H 96-98 N 16 O 13 ). One unit of Polymixin
B Sulfate is equivalent to 0.129 mg of polymixin
Bsulfate(C 55-56 H 96-98 N 16 O 13 .1-2H 2 SO 4 ).
Description Polymixin B Sulfate occurs as a white to yellow-brown
powder.
It is freely soluble in water, and practically insoluble in
ethanol (99.5).
pH The pH of a solution obtained by dissolving 1.0 g of
Polymixin B Sulfate in 50 mL of water is between 5.0 and
7.0.
Phenylalanine Weigh accurately about 0.375 g of Polymixin
B Sulfate, dissolve in 0.1 mol/L hydrochloric acid TS to
make exactly 100 mL. Determine absorbances, A 1 , A 2 , A 3 ,
A 4 and A 5 ,ofthissolutionat252nm,at258nm,at264nm,
at 280 nm and at 300 nm, respectively, as directed under the
Ultraviolet-visible Spectrophotometry, and calculate the
amount of phenylalanine by the following equation: the
amount of phenylalanine calculated on the dried basis is not
less than 9z and not more than 12z.
Amount (z) of phenylalanine
= A 2-0.5A 1 +0.5A 3 -1.8A 4 +0.8A 5
W T
× 9.4787
W T : Amount (g) of the sample, calculated on the dried
basis
Purity Heavy metals—Proceed with 1.0 g of Polymixin B
Sulfate according to Method 2, and perform the test. Prepare
the control solution with 2.0 mL of Standard Lead