A: Infrared Absorption á197Kñ:on undried specimen.

B: The retention time of the major peak in the chromatogram of the Assay preparationcorresponds to that in the chromatogram of the Standard preparation,obtained as directed in the Assay.

C: Prepare a test solution in water containing 15mg per mL.Separately apply 2µLeach of the test solution and a Standard solution of USP Beta Cyclodextrin RSin water containing 5mg per mLto a suitable thin-layer chromatographic plate (see Chromatography á621ñ)coated with a 0.25-mm layer of chromatographic silica gel.Allow the applications to dry,and develop the chromatogram in a solvent system consisting of a mixture of n-propyl alcohol,water,ethyl acetate,and ammonium hydroxide (6:3:1:1)until the solvent front has moved about three-fourths of the length of the plate.Remove the plate from the developing chamber,mark the solvent front,and allow the solvent to evaporate.Locate the spots on the plate by lightly spraying with iodine and potassium iodide TS:the RFvalue of the principal spot obtained from the test solution corresponds to that obtained from the Standard solution.

D: Mix 0.2g with 2mLof iodine TS,warm in a water bath to dissolve the test specimen,and allow to stand at room temperature:a yellow-brown precipitate is formed.

Test solution: 10mg per mL,in water.

Change to read:

Mobile phase— Prepare a filtered and degassed mixture of acetonitrile and water (65:35).Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).

Internal standard solution— Dissolve 2.0g of glycerol in water contained in a 100-mLvolumetric flask,dilute with water to volume,and mix.Pass through a 0.45-µm membrane filter.Use fresh,or store in a freezer,thaw in hot water,and mix.

Standard preparation— Dissolve an accurately weighed quantity of USP Beta Cyclodextrin RSin water,and dilute quantitatively with water to obtain a solution having a known concentration of about 10mg per mL.Use fresh,or store in a freezer,thaw in hot water,and mix.Mix 1.0mLof this solution with 1.0mLof Internal standard solution.

System suitability preparation— Prepare a solution in water containing about 5mg per mLeach of USP Alpha Cyclodextrin RSand USP Beta Cyclodextrin RS.Pass through a 0.45-µm membrane filter.

Assay preparation— Transfer about 1g of Betadex,accurately weighed,to a 100-mLvolumetric flask,dissolve in and dilute with water to volume,and mix.Pass this solution through a 0.45-µm membrane filter.Mix 1.0mLwith 1.0mLof Internal standard solution.

Chromatographic system(see Chromatography á621ñ)— The chromatograph is equipped with a refractive index detector,a 4.6-mm ×25-cm column that contains 10-µm packing L8,and a guard column that contains packing L8.The columns and,if necessary,the detector are maintained at a constant temperature of about 25±2,and the flow rate is about 2.0mLper minute.If the detector or the columns are operated at a temperature other than 25±2,the system also must be shown to meet all system suitability requirements.NF23Chromatograph the System suitability preparation,and record the peak responses as directed for Procedure:the alpha cyclodextrin and beta cyclodextrin peaks exhibit baseline separation,the relative retention times being about 0.8and 1.0,respectively;and the relative standard deviation for replicate injections is not more than 2.0%.

Procedure— Separately inject equal volumes (about 20µL)of the Assay preparationand the Standard preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.Calculate the quantity,in mg,of (C6H10O5)7in the portion of Betadex taken by the formula: in which Cis the concentration,in mg per mL,of anhydrous beta cyclodextrin in the Standard preparation,as determined from the concentration of USP Beta Cyclodextrin RS,corrected for moisture content by a titrimetric water determination;and RUand RSare the peak response ratios of the beta cyclodextrin peak to the internal standard peak obtained from the Assay preparationand the Standard preparation,respectively.