HPLC/MS同时测定双黄连口服液中的4种有效成分

Simultaneous determination of four effective components in Shuanghuanglian Oral Liquid by HPLC/MS

目的建立双黄连口服液(金银花、黄芩、连翘等)中4种有效成分(绿原酸、咖啡酸、黄芩苷和木犀草素)的HPLC/MS同时测定方法。方法采用Zorbax SB C18色谱柱,以含0.2%甲酸0.4 mmol/L醋酸钠的水(A)-甲醇(B)为流动相进行梯度洗脱,0～10 min,35%B;11～25 min,65%B;26～30 min,35%B。体积流量为0.35 mL/min,柱温25℃;在ESI正离子模式下,采用分时段SIM模式:0～7.0 min,m/z 377.4;7.0～12 min,m/z 181.0;12～18 min,m/z 447.1;18～25 min,m/z 287.1。结果绿原酸、咖啡酸、黄芩苷和木犀草素的线性范围分别为0.050～50.0μg/mL(r=0.999 0)、0.030～25.0μg/mL(r=0.998 9)、0.040～300μg/mL(r=0.999 7)和0.010～25.0μg/mL(r=0.999 3);进样精密度RSD值分别为2.0%、2.5%、2.1%和2.4%;加标回收率及其RSD分别为98.5%(RSD 2.7%)、98.8%(RSD 3.7%)、98.5%(RSD1.3%)和99.2%(RSD 2.2%)。结论此方法准确,快速,重复性好,有望为双黄连口服液的质量控制提供依据。

AIM To establish a method for simultaneously determining chlorogenic acid,caffeic acid,baicalin and luteolin in Shuanghuanglian Oral Liquid（Lonicerae japonicac Flos,Scutellariae Radix,Forsythiae Fructus,etc.）.METHODS The analysis was performed on an Agilent Zorbax SB C18 column（3.0 mm × 250 mm,5 μm） using water（A）,containing 0.2% formic acid and 0.4 mmol/L sodium acetate,and methanol（B） as mobile phase,with the gradient elution（0-10 min,35% B;11-25 min,65% B;26-30 min,35% B） at a flow rate of 0.35 mL/min and the column temperature was maintained at 25 ℃.The analytes were detected by ESI-MS in selective ion monitoring mode（0-7.0 min,m/z 377.4;7.0-12 min,m/z 181.0;12-18 min,m/z 447.1;18-25 min,m/z 287.1）.RESULTS The linear ranges were in the ranges of 0.050-50.0 μg/mL（r=0.999 0）,0.030-25.0 μg/mL（r=0.998 9）,0.040-300 μg/mL（r=0.999 7） and 0.010-25.0 μg/mL（r=0.999 3）,respectively,and the RSDs of precision were in the scope 2.0%,2.5%,2.1% and 2.4%,with the average recoveries and their RSDs were 98.5%（RSD 2.7%）,98.8%（RSD 3.7%）,98.5%（RSD 1.3%） and 99.2%（RSD 2.2%） for chlorogenic acid,caffeic acid,baicalin and luteolin,respectively.CONCLUSION The method is accurate,sensitive,rapid,and suitable for the quality control of Shuanghuanglian Oral Liquid.