A:Thin-Layer Chromatographic Identification Test ¨¢201ñ¡ª

B: The retention times of the peaks for chicoric and caftaric acids in the chromatogram of the Test solutioncorrespond to those in the chromatogram of Standard solution 1,as obtained in the test for Content of total phenols.

Change to read:

Solvent,Solution A,Solution B,Mobile phase,Standard solution 2,and Chromatographic system¡ª Proceed as directed for Content of total phenolsunder Echinacea angustifolia.

Standard solution 1¡ª Dissolve an accurately weighed quantity of USP Powdered Echinacea purpureaExtract RSin Solvent,shaking for 1minute,and dilute with Solventto obtain a solution having a known concentration of about 5mg per mL.Pass through a membrane filter having a 0.45-µm or finer porosity.

Test solution¡ª Transfer about 60mg of Powdered Extract,accurately weighed,to a suitable round-bottom flask equipped with a condenser.USP28Add 25mLof Solvent,and heat under reflux while shakingUSP28by mechanical means for 15minutes.Centrifuge,or pass through a membrane filter having a 0.45-µm or finer porosity.

Procedure¡ª Proceed as directed in the test for Content of total phenolsunder Echinacea purpurea Root.Calculate the percentage of each relevant component of total phenols in the portion of Powdered Extract taken by the formula:
in which Fis the response factor and is equal to 0.695for chicoric acid,0.881for caftaric acid,1.000for chlorogenic acid,and 2.220for echinacoside;Cis the concentration,in mg per mL,of USP Chlorogenic Acid RSin Standard solution 2;Wis the weight,in mg,of the portion of Powdered Extract taken;riand rSare the peak responses for the relevant analyte obtained from the Test solutionand Standard solution 2,respectively.Calculate the percentage of total phenols in the portion of Powdered Extract taken by adding the individual percentages.

Change to read:

Mobile phase and Standard solution 2¡ª Proceed as directed for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia.

Standard solution 1¡ª Proceed as directed for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia,except to use USP Powdered Echinacea purpureaExtract RSinstead of USP Powdered Echinacea angustifoliaExtract RS.

Test solution¡ª Proceed as directed for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia,except to use Powdered Echinacea purpureaExtract instead of Powdered Echinacea angustifoliaExtract.

Chromatographic system¡ª Proceed as directed for Content of dodecatetraenoic acid isobutylamidesunder Echinacea angustifolia,except to use the Reference Chromatogram for alkamides provided with USP Powdered Echinacea purpureaExtract RSinstead of the Reference Chromatogram provided with USP Powdered Echinacea angustifoliaExtract RS.

Procedure¡ª Proceed as directed in the test for Content of dodecatetraenoic acid isobutylamides under Echinacea angustifolia.Calculate the percentage of dodecatetraenoic acid isobutylamidesUSP28in the portion of Powdered Extract taken by the formula:
in which 1.353is the response factor for 2E,4E-hexadienoic acid isobutylamide;Cis the concentration,in mg per mL,of USP2E,4E-Hexadienoic Acid Isobutylamide RSin Standard solution 2;Wis the weight,in g,of the portion of Powdered Extract taken;riis the sum of the peak responses of the relevant analytes obtained from the Test solution;and rSis the peak response obtained from Standard solution 2.