A: Thin-Layer Chromatographic Identification Test 201—

B: The retention time of the alliin peak in the chromatogram of the Test solution corresponds to that in the chromatogram of the Standard solution, as obtained in the test for Content of alliin.

0.045 M Phosphate buffer, 0.05 M Phosphate buffer, 0.01 M Carboxymethoxylamine hemihydrochloride solution, Derivatization reagent, Mobile phase, Standard solution, and Chromatographic system— Proceed as directed for Content of alliin under Garlic.

Test solution— Transfer about 0.10 g of Powdered Extract, accurately weighed, to a 50-mL volumetric flask, add 30 mL of 0.01 M Carboxymethoxylamine hemihydrochloride solution, and shake until the Powdered Extract is fully dispersed. Dilute with 0.01 M Carboxymethoxylamine hemihydrochloride solution to volume, and mix. Centrifuge, transfer 5 mL of the clear supernatant to a 10-mL volumetric flask, dilute with 0.01 M Carboxymethoxylamine hemihydrochloride solution to volume, and mix. Using a volumetric syringe, transfer 0.1 mL of this solution to a septum-capped vial, add 0.5 mL of the Derivatization reagent, and mix. Allow a reaction time of not less than 2 minutes before injection into the chromatograph.

Procedure— Proceed as directed for Content of alliin under Garlic. Calculate the percentage of alliin in the portion of Powdered Extract taken by the formula: in which C is the concentration, in mg per mL, of USP Alliin RS in the Standard solution; W is the weight, in g, of Powdered Extract taken to prepare the Test solution; and rU and rs are the sums of the peak responses for the alliin diastereomers obtained from the Test solution and the Standard solution, respectively.

0.045 M Phosphate buffer, 0.05 M Phosphate buffer, 0.01 M Carboxymethoxylamine hemihydrochloride solution, Derivatization reagent, Mobile phase, Standard solution, and Chromatographic system— Proceed as directed for Content of alliin under Garlic.

Test solution— Incubate 200 mg of Powdered Extract with 20 mL of water at room temperature for 5 minutes. Immediately after incubation, add 80.0 mL of 0.01 M Carboxymethoxylamine hemihydrochloride solution, mix, and centrifuge. Using a volumetric syringe, transfer 0.1 mL of this solution to a septum-capped vial, add 0.5 mL of the Derivatization reagent, and mix. Allow a reaction time of not less than 2 minutes before injection into the chromatograph.

Procedure— Proceed as directed for Content of alliin under Garlic: the area of the alliin peak obtained from the chromatogram of the Test solution is not more than 1% of the area of the alliin peak obtained from the chromatogram of the Standard solution.