环孢素-USP35
USP 35Official Monographs / Cyclosporine2793 Dissolution ?711?—amino-6-octenoyl-L-α-aminobutyryl-N-methylglycyl-N-methyl-
L-leucyl-L-valyl-N-methyl-L-leucyl) [59865-13-3].
Medium: pH 6.8 Phosphate buffer (see Buffer Solutions under
Solutions in the section Reagents, Indicators, and Solutions); 900? Cyclosporine contains not less than 98.5 per-mL.
cent and not more than 101.5 per cent of Apparatus 1: 100 rpm.
cyclosporine A (C62H111N11O12), calculated on the Time: 30 minutes.
dried basis.
Determine the amount of C3H6N2O2 dissolved by employing
the following method.
Packaging and storage—Preserve in tight, light-resistant pH 6.8 Phosphate buffer, Mobile phase, and Chromatographic containers.
system—Proceed as directed in the Assay.
Standard solution—Quantitatively dissolve an accurately USP Reference standards ?11?—
weighed quantity of USP Cycloserine RS in pH 6.8 Phosphate USP Cyclosporine RS
buffer to obtain a solution having a known concentration of USP Cyclosporine Resolution Mixture RS
about 0.25 mg per mL.This material is a 100:1 mixture of cyclosporine and
cyclosporine U.
Test solution—Use a filtered portion of the solution under
test.Identification—The chromatogram of the Assay preparation
obtained as directed in the Assay exhibits a major peak for Procedure—Separately inject equal volumes (about 10 μL) of
cyclosporine, the retention time of which corresponds to that the Standard solution and the Test solution into the chromato-
exhibited in the chromatogram of the Standard preparation, as graph, record the chromatograms, and measure the peak re-
obtained in the Assay.
sponses for cycloserine. Calculate the quantity, in mg, of cyc-
loserine (C3H6N2O2) dissolved by the formula:Loss on drying ?731?—Dry about 100 mg, accurately
weighed, in a capillar y-stoppered bottle in vacuum at a pressure 900C(r U/r S)not exceeding 5 mm of mer cury at 60° for 3 hours: it loses not
more than 2.0% of its weight.
in which C is the concentration, in mg per mL, of USP Cycloser-Heavy metals, Method II ?231?: 0.002%.
ine RS in the Standard solution; and r U and r S are the peak re-
Related compounds—Using the chromatograms obtained sponses for cycloserine obtained from the Test solution and the
from Standard preparation 2 and the Assay preparation in the Standard solution, respectively.
Assay, calculate the per centage of each impurity by the formula: Tolerances—Not less than 80% (Q) of the labeled amount of
C3H6N2O2 is dissolved in 30 minutes.2000(C/W)(r
i/r S2)
Uniformity of dosage units ?905?: meet the requirements.
in which C is the concentration, in mg per mL, of USP
Loss on drying ?731?—Dry about 100 mg of the contents of
Cyclosporine RS in Standard preparation 2; W is the weight, in Capsules in a capillar y-stoppered bottle in vacuum at 60° for 3
mg, of Cyclosporine taken to prepare the Assay preparation; r i is hours: it loses not more than 1.0% of its weight.
the response of an individual impurity obser ved in the chromat-Assay—
ogram of the Assay preparation; and r S2 is the response of the pH 6.8 Phosphate buffer, Mobile phase, Standard preparation,main cyclosporine peak in the chromatogram obtained from and Chromatographic system—Proceed as directed in the Assay Standard preparation 2: not more than 0.7% of any individual under Cycloserine.impurity is found, and the sum of all such impurities is not Assay preparation—Remove, as completely as possible, the more than 1.5%, any impurities corresponding to less than contents of not fewer than 20 Capsules. T ransfer an accurately0.05% being disregarded.
weighed portion of the powder, equivalent to about 100 mg of Assay—
cycloserine, to a 250-mL volumetric flask, dilute with pH 6.8
Mobile phase—Prepare a mixture of water, acetonitrile, tert-Phosphate buffer to volume, mix, and filter.
butyl methyl ether, and phosphoric acid (520:430:50:1). Make Procedure—Proceed as directed in the Assay under Cycloser-adjustments if necessar y (see System Suitability under Chroma-ine. Calculate the quantity, in mg, of cycloserine (C3H6N2O2) in tography ?621?).
the portion of Capsules taken by the formula:
Diluent—Prepare a mixture of acetonitrile and water (1:1).
250C(r U/r S)Standard preparation 1—Dissolve an accurately weighed
quantity of USP Cyclosporine RS in Diluent to obtain a solution in which the terms are as defined therein.having a known concentration of about 1.25 mg per mL.
Standard preparation 2—Transfer 2.0 mL of Standard prepara-
tion 1 to a 250-mL volumetric flask, dilute with Diluent to vol-
ume, and mix. This solution contains about 0.01 mg of USP
Cyclosporine RS per mL.
Cyclosporine
Assay preparation—Dissolve about 25 mg of Cyclosporine,
accurately weighed, in Diluent, dilute with Diluent to 20.0 mL,
and mix.
Resolution solution—Prepare a solution of USP Cyclosporine
Resolution Mixture RS in Diluent having a concentration of
about 1.25 mg per mL.
Chromatographic system (see Chromatography ?621?)—The
liquid chromatograph is equipped with a 210-nm detector, a
0.25-mm × 1-m stainless steel tube connected to a 4-mm × 25-
cm column that contains 3- to 5-μm packing L1. The tube and C62H111N11O121202.61column are maintained at 80°. The flow rate is about 1.2 mL Cyclo[[(E)-(2S,3R,4R)-3-hydroxy-4-methyl-2-(methylamino)-6-per minute. Chromatograph the Resolution solution, and record octenoyl]-L-2-aminobutyryl-N-methylglycyl-N-methyl-L-leucyl-the responses as directed for Procedure: the cyclosporine U peak L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-and the main cyclosporine peak are resolved from each other. methyl-L-leucyl-N-methyl-L-valyl].Chromatograph Standard preparation 1, and record the re-
[R-[R*,R*-(E)]]-Cyclic(L-alanyl-D-alanyl-N-methyl-L-leucyl-N-sponses as directed for Procedure: the relative standard deviation methyl-L-leucyl-N-methyl-L-valyl-3-hydroxy-N,4-dimethyl-L-2-for replicate injections is not more than 1.0%. Chromatograph
2794Cyclosporine / Official Monographs USP 35
Standard preparation 2, and record the responses as directed for Test solution—Filter a portion of the solution under test. Procedure: the relative standard deviation for replicate injections Transfer 5.0 mL of the filtrate to a 10-mL volumetric flask, dilute is not more than 10%.with acetonitrile to volume, and mix.
Procedure—[NOTE—Use peak areas where peak responses are Chromatographic system (see Chromatography ?621?)—The indicated.] Separately inject equal volumes (about 20 μL) of liquid chromatograph is equipped with a 210-nm detector and Standard preparation 1, Standard preparation 2, and the Assay a 4.6-mm × 25-cm column that contains packing L1 and is preparation into the chromatograph, record the chromato-maintained at a constant temperature of about 80°. The flow grams, and measure the peak responses. Calculate the per cent-rate is about 2 mL per minute. Chromatograph the Standard age of cyclosporine A (C62H111N11O12) in the Cyclosporine taken solution, and record the peak areas as directed for Procedure:
by the formula:the column efficiency is not less than 700 theoretical plates;
and the relative standard deviation for replicate injections is not (CP/10U)(r U/r S)more than 2.0%.
Procedure—Separately inject equal volumes (about 10 μL) of in which C is the concentration, in mg per mL, of USP the solution estimated to contain 0.1 mg of cyclosporine per Cyclosporine RS in Standard preparation 1; P is the specified mL, or 40 μL of the solution estimated to contain 0.025 mg of purity, in μg per mg, of USP Cyclosporine RS; U is the concen-cyclosporine per mL) of the Standard solution and the Test solu-tration, in mg per mL, of specimen in the Assay preparation;tion into the chromatograph, record the chromatograms, and and r U and r S are the main cyclosporine peak responses ob-measure the areas for the major peaks. Calculate the quantity, tained from the Assay preparation and Standard preparation 1,in mg, of C
62H111N11O12 dissolved by the formula: respectively.
2000C(r U/r S)
in which C is the concentration, in mg per mL, of USP
Cyclosporine RS in the Standard solution; and r U and r S are the Cyclosporine Capsules cyclosporine peak areas obtained from the Test solution and the
Standard solution, respectively.
? Cyclosporine Capsules contain not less than Tolerances—Not less than 80% (Q) of the labeled amount of 90.0 percent and not more than 110.0 per cent C
62H111N11O12 is dissolved in 90 minutes.
of the labeled amount of cyclosporine Uniformity of dosage units ?905?: meet the requirements. (C62H111N11O12).Water, Method I ?921?—For Capsules that contain powder, not
more than 3.5% is found, using finely ground Capsule Packaging and storage—Preserve in tight containers, and contents.
store at controlled room temperature.Assay—
Identification—The retention time of the major peak in the
WHERE CAPSULES CONTAIN LIQUID—
chromatogram of the Assay preparation corresponds to that in
Mobile phase and Chromatographic system—Proceed as di-the chromatogram of the Standard preparation, as obtained in
rected in the Assay under Cyclosporine Injection.
the Assay.
Standard preparation—Dissolve an accurately weighed quan-Dissolution ?711?—
tity of USP Cyclosporine RS in dehydrated alcohol to obtain a WHERE CAPSULES CONTAIN LIQUID—solution having a known concentration of about 1 mg per mL.
Medium: water; 500 https://www.wendangku.net/doc/f02225201.html,e this solution promptly after preparation.
Apparatus 2: 50 rpm.Assay preparation—Using a sharp blade, carefully cut open Time: 15 minutes.not fewer than 20 Capsules, and with the aid of dehydrated
alcohol transfer the contents of the Capsules to a suitable volu-Procedure—Place 1 Capsule in each vessel, and allow the
metric flask. Wash the blade with dehydrated alcohol, and Capsule to sink to the bottom of the vessel before starting rota-
transfer the washings to the volumetric flask. Dilute the con-tion of the blade. Obser ve the Capsules, and record the time
tents of the volumetric flask with dehydrated alcohol to volume, taken for each Capsule shell to rupture.
and mix. Quantitatively dilute an accurately measured volume Tolerances—The requirements are met if all of the Capsules
of this solution with dehydrated alcohol to obtain a solution tested rupture in not more than 15 minutes. If 1 or 2 of the
having a concentration of about 1 mg of cyclosporine per mL. Capsules rupture in more than 15 but not more than 30 min-
Procedure—Separately inject equal volumes (about 20 μL) of utes, repeat the test on 12 additional Capsules. Not more than
the Standard preparation and the Assay preparation into the
2 of the total of 18 Capsules tested rupture in more than 15
chromatograph, record the chromatograms, and measure the but not more than 30 minutes.
areas for the major peaks. Calculate the quantity, in mg, of WHERE CAPSULES CONTAIN POWDER—cyclosporine (C
62H111N11O12) in each Capsule taken by the Medium: 0.1 N hydrochloric acid containing 0.5% of so-formula:
dium lauryl sulfate; 1000 mL.
Apparatus 1: 150 rpm.(L/D)(CP/1000)(r U/r S)
Time: 90 minutes.
in which L is the labeled quantity, in mg, of cyclosporine in Determine the amount of C62H111N11O12 dissolved by employ-
each Capsule taken; D is the concentration, in mg per mL, of ing the following method.
the Assay preparation, based on the labeled quantity of Mobile phase—Prepare a filtered and degassed mixture of ac-
cyclosporine in the Capsules taken and the extent of dilution; C etonitrile, water, methanol, and phosphoric acid
is the concentration, in mg per mL, of USP Cyclosporine RS in (900:450:50:0.5). Make adjustments if necessar y (see System
the Standard preparation; P is the purity, in μg per mg, of USP Suitability under Chromatography ?621?).
Cyclosporine RS; and r U and r S are the peak areas obtained from Standard solution—Quantitatively dissolve an accurately the Assay preparation and the Standard preparation, respectively. weighed quantity of USP Cyclosporine RS in Dissolution Medium
WHERE CAPSULES CONTAIN POWDER—
to obtain a solution having a known concentration of about
Mobile phase—Prepare a filtered and degassed mixture of ac-0.001L mg per mL, L being the labeled quantity, in mg, of
etonitrile, water, methanol, and phosphoric acid
cyclosporine in each Capsule. T ransfer 25.0 mL of this solution
(605:400:50:0.5). Make adjustments if necessar y (see System
to a 50-mL volumetric flask, dilute with acetonitrile to volume,
Suitability under Chromatography ?621?).
and mix. This solution contains about 0.0005L mg of USP
Cyclosporine RS per mL.