液相色谱-质谱串联法测定清血内消丸中马兜铃酸A含量

目的:建立清血内消丸中马兜铃酸A的限量检查法.方法:采用1％碳酸钠溶液多次加热溶解样品,用三氯甲烷多次提取,弃去三氯甲烷层,碱水层用稀盐酸调pH,加三氯甲烷提取多次,收集合并三氯甲烷层,蒸干,残渣加甲醇溶解,精密吸取1 mL,置20 mL量瓶,加乙腈-水(40∶60)稀释至刻度,以水(0.1％甲酸+5 mmol· L-1甲酸铵)/(％)和乙腈(含5％水,0.1％甲酸+5 mmol· L-1甲酸铵)/(％)为流动相梯度洗脱,应用高效液相色谱/电喷雾质谱多离子反应监测模式(MRM)测定.结果:马兜铃酸A在11.21 ～299.0 pg进样量与马兜铃酸A峰面积呈良好的线性关系(r=0.999 5),检测限0.1μg·L-1,定测限0.3 μg·L-1,平均回收率119.20％,RSD4.0％(n=9).结论:方法准确,快速,简便,重复性好,干扰少,特异性好,能满足马兜铃酸A的痕量残留的检测要求.

Determination of Aristolochic Acid A in Qingxueneixiao Pills by LC-MS-MS

Objective: To establish a method to examine the limited content of aristolochic acid A in Qingxueneixiao Pills. Method: 1% sodium carbonate solution is repeatedly heated to dissolve the sample, extracted with chloroform for several times. Discard the chloroform layer, adjusted the pH of the alkaline layer with dilute hydrochloric acid, and extracted with chloroform again for several times. Collect and evaporate the chloroform layer. The residue was dissolved in methanol, 1 mL of which was diluted to 20 mL, eluted with a gradient composed by acetonitrile-water (40: 60), water (0.1% formic acid+5 mmol·L^-1 ammonium formate )/( %) and acetonitrile (containing 5% water, 0.1% formic acid+5 mmol·L^-1 ammonium formate )/( %), assayed in high performance liquid chromatography/electrospray ionization mass spectrometry multiple reaction monitoring (MRM) mode. Result: Aristolochic acid A was linear int the range of 11.21-299.0 pg (r=0.999 5).The average recovery was 119.2% (n=9) and the RSD was 4.0%. Conclusion: This method is accurate, rapid, simple, reproducible, less interference and specific, appropriate to the trace detection for the Qingxueneixiao Pills.