HPLC-DAD/MS法测定抗风湿中成药中非法添加10种抗风湿化学成分

Determination of Ten Antirheumatic Components Illegally Added in Chinese Patent Medicine by HPLC-DAD/MS

目的:建立检测抗风湿中成药中非法添加10种抗风湿化学成分的HPLC-DAD方法,并用MS/MS进行了化合物的归属分析。方法:用C18色谱柱(4.6 mm×250 mm,5μm),以0.02 mol·L^(-1)乙酸铵(含0.05%乙酸)水溶液为流动相A,以乙腈为流动相B,梯度洗脱,波长240 nm,流速1.0 mL·min^(-1),进行HPLC定性定量分析;再采用串联四极杆质谱的MRM模式检测,进行化合物归属分析,检测添加在中成药中的对乙酰氨基酚、甲氧苄啶、氨基比林、吡罗昔康、萘普生、双氯芬酸钠、醋酸泼尼松、吲哚美辛、保泰松、布洛芬10种化学成分。结果:10种抗风湿性化学成分液相检测的线性范围宽,相关性好,r≥0.999 2,方法精密度符合要求;回收率在98.0%~100.9%之间;精密度、稳定性均符合要求;检测限为0.05~0.5μg·mL^(-1);定量限为0.1~1.8μg·mL^(-1)。结论:本方法专属性强,操作简单,快捷,可作为中成药中非法添加以上10种抗风湿性化学成分的有效检测方法。

Objective： To establish an HPLC-DAD method for determination of 10 antirheumatic components illegally added in Chinese patent medicines with MS/MS attribution analysis for these components. Methods： The test was performed in C18 column（4.6mm×250mm,5μm） under the gradient elution with mobile phase A（0.02mol·L^-1 ammonium acetate 5% acetic acid solution） and mobile phase B （acetonitrile）. The flow rate was 1. 0mL·min^-1, and the detection wavelength was 240 nm. Multiple-reaction monitoring（MRM） mode of tandem quadru- pole mass spectrometry was adopted to identify paracetamol, trimethoprim, aminopyrine, piroxieam, naproxen, diclofenac sodium, prednisone acetate, indomethacin, phenylbutazone and ibuprofen. Results： There were the wide linear ranges with r≥0. 999 7 for 10 antirheumatic components. The precision of the method met the criteria, and the recoveries of 10 components fell into 98.0% to 100. 9%. The precision and stability data met the requirements. The limits of detection fell into 0. 05 μg·mL^-1 to 0. 5 μg·mL^-1, and the limits of quantitation fell into 0. 1μg·mL^-1 to 1.8 μg·mL^-1. Conclusion： The established method has the good specificity, and it is simple. It can be applicable to determination of 10 antirheumatic components illegally added in Chinese patent medicines.