A: The retention times of the major peaks in the chromatogram of the Assay preparationcorrespond to those in the chromatogram of the Standard preparation,both relative to the internal standard as obtained in the Assay.

B: Using a sharp blade,carefully open 1Capsule and transfer the entire contents into a 250-mLflask.Inspect the encapsulated liquid to ensure that crystallization or agglomeration of the drug substance has not taken place.Add 5mLof water to the flask,and swirl to dissolve.Add 10mLof p-dimethylaminobenzaldehyde TS:a blue color develops within 2minutes and persists for not less than 10minutes.

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 note if there is membrane formation.Record the time that each Capsule ruptures.

Tolerances— All of the Capsules tested rupture in not more than 15minutes.If 1or 2Capsules fail to rupture in 15minutes but rupture in 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.

Mobile phase— Prepare a filtered and degassed mixture of water,acetonitrile,and triethylamine (32:18:1).Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).

Internal standard solution— Dissolve an accurately weighed quantity of m-chloroacetanilide in acetonitrile to obtain a solution having a known concentration of about 0.12mg per mL.

Tartaric acid solution— Dissolve an accurately weighed quantity of tartaric acid in water to obtain a solution having a known concentration of about 5.6mg per mL.

Standard preparation— Transfer about 25.0mg of USP Ergoloid Mesylates RS,accurately weighed,to a 100-mLvolumetric flask,dissolve in and dilute with Internal standard solutionto volume,and mix.Transfer 40.0mLof this solution to a 200-mLvolumetric flask,add 40.0mLof Tartaric acid solution,and mix.

Assay preparation— Pipet 40.0mLof Tartaric acid solutioninto a 200-mLvolumetric flask,and heat in a water bath maintained at 50.Add 10Capsules (or the equivalent of 10mg of ergoloid mesylates),and shake the flask by mechanical means for 10minutes or until the gelatin has dissolved.Pipet 40.0mLof Internal standard solutioninto the flask,and shake for an additional 10minutes.Remove the flask from the bath and cool to room temperature.Transfer about 20mLof the solution to a 30-mLcentrifuge tube,and centrifuge at 10,000rpm for 60minutes.Filter a portion of the supernatant through a filter having a porosity of 0.45µm,discarding the first 5mLof the filtrate.Use the remainder of the filtrate as the Assay preparation.

Chromatographic system (see Chromatography á621ñ)—The liquid chromatograph is equipped with a 280-nm detector and an 8-mm ×10-cm column that contains packing L1.The flow rate is about 2mLper minute.Chromatograph the Standard preparation,and record the peak responses as directed under Procedure:the relative retention times for m-chloroacetanilide,dihydroergocornine,dihydro-a-ergocryptine,dihydroergocristine,and dihydro-b-ergocryptine are about 1.0,1.4,1.8,2.2,and 2.8,respectively;the column efficiency determined for the dihydro-b-ergocryptine peak is not less than 950theoretical plates;the tailing factor for dihydro-b-ergocryptine is not more than 2.5and that for dihydroergocornine is not more than 2.0;the resolution,R,between dihydro-a-ergocryptine and dihydroergocristine is not less than 1.35;and the relative standard deviation of the sum of the four ergoloid mesylate peaks for replicate injections is not more than 2.0%.

Procedure— Separately inject equal volumes (about 50µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.Calculate the total quantity,in mg,of ergoloid mesylates in the portion of Capsules taken by the formula: in which Cis the concentration,in mg per mL,of USP Ergoloid Mesylates RSin the Standard preparation,and SRUand SRSare sums of the ratios of the peak responses of the individual alkaloids to the peak response of the internal standard obtained from the Assay preparationand the Standard preparation,respectively.Calculate the percentage of each of the individual alkaloids in the portion of Capsules taken by the formula: in which (MW)Uis the molecular weight of the individual alkaloid.