二十五味绿绒蒿丸中马兜铃酸A的分析___

Objective:To establish the TLC identification and HPLC determination methods of aristolochic acid A in Ershiwuwei Luronghao pills.Methods:TLC and HPLC were adopted to identify and determine aristolochic acid A in the pills.TLC:Using silica gel G mixed with sodium carboxymethylcellulose as the coating substance and toluene-acetoacetate-water-formic acid(20∶10∶1∶1) upper part solution as the mobile phase.After developing about 12 cm from the base line，removal of the plate,dry in air and examine under ultraviolet light(365 nm).HPLC conditions:Using Phenomenex Luna C₁₈(2)(4.6 mm×150 mm,5 μm) column as analytic column and 0.05 mol·L^-1 sodium dihydrogen phosphate(containing 1 mL·L^-1 phosphoric acid)-acetonitrile(62∶38) as the mobile phase, flow rate at 1.0 mL·min^-1；As detector spectrophotometer set at 315 nm,column temperature at 35 ℃.Results:Aristolochic acid A could be examined by TLC and determined by HPLC.The dark spot in chromatogram obtained with the test solution corresponds in position and colour to the spot in chromatogram obtained with the reference solution.The linear range of aristolochic acid A was 0.10-0.61 μg(r=0.9997),the average recovery(n=9)was 96.20%.Conclusion:The spot on the TLC plate were clearly，and the negative contrast was not disturbed；The methods is simple,accurate and can be used for the quality control of Ershiwuwei Luronghao pills.