A: Ultraviolet Absorption 197U—

B: Thin-Layer Chromatographic Identification Test 201—

Mobile phase and Chromatographic system— Prepare as directed in the Assay.

Test solution— Transfer about 100 mg of Cyclandelate, accurately weighed, to a 100-mL volumetric flask, dissolve in and dilute with Mobile phase to volume, and mix.

Standard solution— Pipet 3.0 mL of the Test solution into a 100-mL volumetric flask, dilute with Mobile phase to volume, and mix.

Procedure— Separately inject equal volumes (about 10 µL) of the Test solution and the Standard solution into the chromatograph, record the chromatograms, and measure the areas for all the peaks. Allow the chromatogram of the Test solution to run for a period of time that is about 3 times the retention time of cyclandelate. The total area of all the peaks from the Test solution, other than the peak obtained from cyclandelate, is not greater than the peak area of cyclandelate obtained from the Standard solution: not more than 3.0% of total impurities is found.

(Official January 1, 2007)

Mobile phase— Prepare a filtered and degassed mixture of acetonitrile and water (4:1). Make adjustments if necessary (see System Suitability under Chromatography 621).

Resolution solution— Dissolve accurately weighed quantities of USP Cyclandelate RS and dicyclohexyl phthalate in Mobile phase to obtain a solution having known concentrations of about 0.2 mg per mL and 0.08 mg per mL, respectively.

Standard preparation— Dissolve an accurately weighed quantity of USP Cyclandelate RS in Mobile phase to obtain a solution having a known concentration of about 1.0 mg per mL. Pipet 10.0 mL of this solution into a 50-mL volumetric flask, dilute with Mobile phase to volume, and mix.

Assay preparation— Transfer about 100 mg of Cyclandelate, accurately weighed, to a 100-mL volumetric flask, dissolve in and dilute with Mobile phase to volume, and mix. Pipet 10.0 mL of this solution into a 50-mL volumetric flask, dilute with Mobile phase to volume, and mix.

Chromatographic system (see Chromatography 621)— The liquid chromatograph is equipped with a 228-nm detector and a 4.0-mm × 15-cm column that contains packing L1. The flow rate is about 1.0 mL per minute. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the resolution, R, between cyclandelate and dicyclohexyl phthalate is not less than 7. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the relative standard deviation for replicate injections is not more than 2.0%.

Procedure— Separately inject equal volumes (about 10 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in mg, of C17H24O3 in the portion of Cyclandelate taken by the formula: in which C is the concentration, in mg per mL, of USP Cyclandelate RS in the Standard preparation; and rU and rS are the peak responses obtained from the Assay preparation and the Standard preparation, respectively.