HPLC/MS测定桂林西瓜霜喷剂中的4种有效成分

Simultaneous determination of four effective components in Guilin Watermelon Frost by HPLC-MS

目的：建立一种快速灵敏的同时测定桂林西瓜霜喷剂（西瓜霜，黄连，黄芩等）中的有效成分苦参碱、小檗碱、黄芩苷和靛玉红的液相色谱／质谱（HPLC／MS）方法。方法：采用Zorbax SB C18（3．0mm×250mm，5μm）色谱柱，以含0．5％甲酸的水（A）-甲醇（B）体系为流动相进行线性梯度洗脱，梯度洗脱程序为：0—2min，60％B；2～5min，60％-90％B；5min至结束，90％B；流速：0．40mL／min；在ESI正离子模式下，采用选择性离子监测方法：0—3min，m／z249；3—6min，m／z336；6～10min，m／z447；10～16min，m／z263。苦参碱在0．020—10μg／mL，小檗碱在0．010～40．0μg／mL，黄芩苷在0．036～50．0μg／mL，靛玉红在0．040～4．00μg／mL范围内峰面积与浓度呈良好的线性关系；检出限分别为0．005，0．001，0．006，0．010μg／mL。4种成分的加样回收率均在96％～101％之间，相对标准偏差RSD小于3％。结论：方法快捷，准确，重现性好。可为桂林西瓜霜喷剂的质量控制提供参考依据。

AIM ： To establish an HPLC-MS method for simultaneously determining four effective components in Guilin Watermelon Frost （ Mirabilitum Praeparatum, Rhizoma Coptidis, Radix Scutellariae, etc. ）. METHODS ： The sample was extracted with methanol containing 20% chloroform under ultrasonication. The HPLC separation was performed on Zorbax SB Cls （3.0 mm ×250 mm, 5 μm） column using water including 0.5% formic acid （A）-methanol （B） as mobile phase, with the gradient elution （0-2 min, 60% B; 2-5 min, 60%±90% B; 5 rain to the end, 90%B）, at a flow rate of 0.40 mL/min. The compounds were analyzed by ESI-MS under ion monito- ring mode（0-3 min, m/z 249; 3-6 min, m/z 336; 6-10 min, m/z447; 10-16 min, m/z 263）. RESULTS： The linear ranges were 0. 020-10.0 μg/mL, 0. 010-40.0 μg/mL, 0. 036-50.0 μg/mL and 0. 040-4.00 μg/mL for matrine, berberine, baicalin and indirubin, with detection limits of 0. 005, 0.001, 0. 006 and 0. 010 μg/mL, respectively. The average recoveries ranged from 96% to 101% with all relative standard deviations less than 3%.CONCLUSION： The method is rapid, accurate and suitable for the quality control of the four effective components in Guilin Watermelon Frost.