LC-MS/MS法测定藏药十三味红花丸中羟基红花黄色素A的含量

目的:建立测定藏药十三味红花丸中羟基红花黄色素A含量的方法。方法:样品经甲醇-水(80∶20,V/V)提取,0.22μm微孔滤膜滤过后采用液相色谱-串联质谱(LC-MS/MS)法测定其中羟基红花黄色素A的含量。色谱柱为AgilentZorbaxSB-C18(50mm×2.1mmI.D.,3.5μm),流动相为水(含0.1%甲酸)-乙腈(含0.1%甲酸),梯度洗脱;离子源为电喷雾离子化源(ESI源),离子源温度为500℃,以多反应监测(MRM)方式分别监测离子对m/z611.2→491.2(羟基红花黄色素A)和m/z268.9→159.0(内标染料木素)。结果:羟基红花黄色素A的检测浓度在10～1000ng·mL-1范围内同其与内标的峰面积比值呈良好线性关系(r=0.9989);平均加样回收率在93.8%～106.2%范围内,RSD均<4.90%(n=9)。结论:该方法快速、灵敏,结果准确,适用于藏药十三味红花丸的质量控制。

Content Determination of Hydroxysafflor Yellow A in Tibetan Medicine Shisanwei Honghua Pills by LC-MS/ MS

OBJECTIVE： To determine the content of hydroxysafflor yellow A in Tibetan medicine Shisanwei honghua pills. METHODS： The samples were firstly extracted with methanol-water solution （80：20, V/V）, and then filtered through 0.22 μm mi- crofilter. The content of hydroxysafflor yellow A was determined by LC-MS/MS. The determination was performed on Agilent Zorb- ax SB-C18 （50 mmx 2.1 mm I.D., 3.5μm） column with mobile phase consisted of water （containing 0.1% formic acid ） -acetonitrile （containing 0.1% formic acid） with gradient elution. The mass spectrometer was operated in the negative ionization electrospray （ESI） mode using multiple reaction monitoring （MRM） for analysis of hydroxysafflor yellow A at 500 ℃. The transitions of m/z 611.2→491.2 （hydroxysafflor yellow A） and m/z 268.9→159.0 （genistein） were used to quantify hydroxysattlor yellow A and ge- nistein. RESULTS： The linear range of hydroxysafflor yellow A was 10-1 000 ng ·mL^-1（r=0.998 9） ; Average recoveries were 93.8% -106.2% （RSD〈4.90%, n=9）. CONCLUSION： The method is rapid, sensitive and accurate, and suitable for the quality control of Tibetan medicine Shisanwei honghua pills.