Medium: water;500mL.

Apparatus 2: 50rpm.

Time: 15minutes.

Procedure— Place 1Capsule in each vessel,and allow the Capsule to sink to the bottom of the vessel before starting rotation of the blade.Observe the Capsules,and record the time taken for each Capsule shell to rupture.

Tolerances— The requirements are met if all of the Capsules tested rupture in not more than 15minutes.If 1or 2of the Capsules rupture in more than 15but not more than 30minutes,repeat the test on 12additional Capsules.Not more than 2of the total of 18Capsules tested rupture in more than 15but not more than 30minutes.

Medium: 0.1Nhydrochloric acid containing 0.5%of sodium lauryl sulfate;1000mL.

Apparatus 1: 150rpm.

Time: 90minutes.

Mobile phase— Prepare a filtered and degassed mixture of acetonitrile,water,methanol,and phosphoric acid (900:450:50:0.5).Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).

Standard solution— Quantitatively dissolve an accurately weighed quantity of USP Cyclosporine RSin Dissolution Mediumto obtain a solution having a known concentration of about 0.001Lmg per mL,Lbeing the labeled quantity,in mg,of cyclosporinein each Capsule.Transfer 25.0mLof this solution to a 50-mLvolumetric flask,dilute with acetonitrile to volume,and mix.This solution contains about 0.0005Lmg of USP Cyclosporine RSper mL.

Test solution— Filter a portion of the solution under test.Transfer 5.0mLof the filtrate to a 10-mLvolumetric flask,dilute with acetonitrile to volume,and mix.

Chromatographic system (see Chromatography á621ñ)— The liquid chromatograph is equipped with a 210-nm detector and a 4.6-mm ×25-cm column that contains packing L1and is maintained at a constant temperature of about 80.The flow rate is about 2mLper minute.Chromatograph the Standard solution,and record the peak areas as directed for Procedure:the column efficiency is not less than 700theoretical plates;and the relative standard deviation for replicate injections is not more than 2.0%.

Procedure— Separately inject equal volumes (about 10µL)of the solution estimated to contain 0.1mg of cyclosporine per mL,or 40µLof the solution estimated to contain 0.025mg of cyclosporine per mL)of the Standard solutionand the Test solutioninto the chromatograph,record the chromatograms,and measure the areas for the major peaks.Calculate the quantity,in mg,of C62H111N11O12dissolved by the formula: in which Cis the concentration,in mg per mL,of USP Cyclosporine RSin the Standard solution;and rUand rSare the cyclosporine peak areas obtained from the Test solutionand the Standard solution,respectively.

Tolerances— Not less than 80%(Q)of the labeled amount of C62H111N11O12is dissolved in 90minutes.

Mobile phase and Chromatographic system— Proceed as directed in the Assayunder Cyclosporine Injection.

Standard preparation— Dissolve an accurately weighed quantity of USP Cyclosporine RSin dehydrated alcohol to obtain a solution having a known concentration of about 1mg per mL.Use this solution promptly after preparation.

Assay preparation— Using a sharp blade,carefully cut open not fewer than 20Capsules,and with the aid of dehydrated alcohol transfer the contents of the Capsules to a suitable volumetric flask.Wash the blade with dehydrated alcohol,and transfer the washings to the volumetric flask.Dilute the contents of the volumetric flask with dehydrated alcohol to volume,and mix.Quantitatively dilute an accurately measured volume of this solution with dehydrated alcohol to obtain a solution having a concentration of about 1mg of cyclosporine per mL.

Procedure— Separately inject equal volumes (about 20µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the areas for the major peaks.Calculate the quantity,in mg,of cyclosporine (C62H111N11O12)in each Capsule taken by the formula: in which Lis the labeled quantity,in mg,of cyclosporine in each Capsule taken;Dis the concentration,in mg per mL,of the Assay preparation,based on the labeled quantity of cyclosporine in the Capsules taken and the extent of dilution;Cis the concentration,in mg per mL,of USP Cyclosporine RSin the Standard preparation;Pis the purity,in µg per mg,of USP Cyclosporine RS;and rUand rSare the peak areas obtained from the Assay preparationand the Standard preparation,respectively.

Mobile phase— Prepare a filtered and degassed mixture of acetonitrile,water,methanol,and phosphoric acid (605:400:50:0.5).Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).

Diluting solvent— Prepare a mixture of acetonitrile,tetrahydrofuran,and dehydrated alcohol (9:5:4).

Standard preparation— Transfer about 25mg of USP Cyclosporine RS,accurately weighed,to a 25-mLvolumetric flask.Add 2.5mLof water,and sonicate for 10minutes.Add about 10mLof Diluting solvent,sonicate for 5minutes,dilute with Diluting solventto volume,and mix.

Assay stock preparation— Transfer the contents of 20Capsules to a volumetric flask of such capacity,V,in mL,to make a final concentration of 10mg of cyclosporine per mL.Add 0.1VmLof water to the flask,and sonicate for 10minutes.Add 0.4VmLof Diluting solventto the flask,and sonicate for 5minutes.Dilute with Diluting solventto volume,and mix.

Assay preparation— Transfer 5.0mLof Assay stock preparationto a 50-mLvolumetric flask,add 5mLof water,dilute with Diluting solventto volume,and mix.

Chromatographic system (see Chromatography á621ñ)— The liquid chromatograph is equipped with a 210-nm detector and a 4.6-mm ×25-cm column that contains packing L13and is maintained at a constant temperature of about 70.The flow rate is about 2mLper minute.Chromatograph the Standard preparation,and record the peak areas as directed for Procedure:the column efficiency is not less than 700theoretical plates;the tailing factor is not more than 1.5;and the relative standard deviation for replicate injections is not more than 2.0%.

Procedure— Separately inject equal volumes (about 20µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the areas for the major peaks.Calculate the quantity,in mg,of cyclosporine (C62H111N11O12)in each Capsule taken by the formula: in which Cis the concentration,in mg per mL,of USP Cyclosporine RSin the Standard preparation;Vis the volume,in mL,of the volumetric flask used to prepare the Assay stock preparation;and rUand rSare the cyclosporine peak areas obtained from the Assay preparationand the Standard preparation,respectively.