参柏抑菌洗剂中6种活性成分的含量测定

Simultaneous determination of 6 constituents in Shenbai Lotion

目的 建立高效液相色谱串联三重四级杆质谱法(HPLC-MS/MS)同时测定参柏抑菌洗剂中6种成分的含量。方法 采用HPLC-MS/MS分析方法,色谱柱为Shim-pack GIST-HP C_(18)(2.1 mm×100.0 mm,3μm),流动相为甲醇(A)-体积分数0.1%甲酸水溶液(B),梯度洗脱,流速0.3 mL/min,进样量5μL,柱温35℃。质谱条件:采用电喷雾电离源(ESI),正、负离子切换扫描模式,多反应监测模式(MRM)检测6种成分(包括氧化苦参碱、小檗碱、黄柏碱、蛇床子素、罗丹明B、落新妇苷)的含量。结果 参柏抑菌洗剂中氧化苦参碱、小檗碱、黄柏碱、蛇床子素、罗丹明B、落新妇苷分别在20.75~622.50μg/L、24.96~748.80μg/L、8.25~247.50μg/L、2.80~84.00μg/L、0.42~12.60μg/L、31.50~945.00μg/L范围内线性关系良好(均r≥0.999 0),精密度、重复性、稳定性试验的相对标准偏差(RSD)均<3%;平均加样回收率为98.58%~100.32%(RSD<4%)。6批参柏抑菌洗剂样品中氧化苦参碱、小檗碱、黄柏碱、蛇床子素、罗丹明B、落新妇苷的含量范围分别为69.862~73.062μg/g,120.997~124.097μg/g,37.323~39.111μg/g,11.425~12.258μg/g,0.742~0.822μg/g,101.286~105.677μg/g。结论 该实验建立的HPLC-MS/MS方法简便、灵敏、高效,可用于参柏抑菌洗剂中多种活性成分的测定和质量控制。

Objective To establish high performance liquid chromatography tandem triple quadrupole mass spectrometry(HPLC-MS/MS)method for simultaneous determination of 6 chemical constituents in Shenbai Lotion.Methods HPLC-MS/MS was used and the chromatographic conditions were as follows:Shim-pack GIST-HP C18 column(2.1 mm×100.0 mm,3μm),mobile phase was consisted of methanol(A)-water(0.1%formic acid,B)at a flow rate of 0.3 mL/min with gradient elution,injected 5μL to HPLC-MS/MS,and the column temperature was 35℃.Mass spectrometer conditions:triple quadrupole mass spectrometer equipped with electrospray ionization source(ESI)was used in positive and negative ion mode,and multiple reaction monitoring(MRM)for the detection of 6 constituents(including oxymatrine,berberine,phellodendron,osthole,rhodamine B and astilbin).Results Under the optimized conditions,oxymatrine,berberine,phellodendron,osthole,rhodamine B and astilbin all showed good linear relationships in the ranges of 20.75-622.50μg/L,24.96-748.80μg/L,8.25-247.50μg/L,2.80-84.00μg/L,0.42-12.60μg/L and 31.50-945.00μg/L(all r≥0.9990),relative standard deviation(RSD)of stability,precision and reproducibility tests was all<3%,and average recoveries were 98.58%-100.32%(RSD<4%).The average contents of oxymatrine,berberine,phellodendron,osthole,rhodamine B and astilbin in 6 batches of Shenbai Lotion were 69.862-73.062μg/g,120.997-124.097μg/g,37.323-39.111μg/g,11.425-12.258μg/g,0.742-0.822μg/g and 101.286-105.677μg/g.Conclusion This simple,sensitive and efficient method could be used for the rapid determination of main active components in Shenbai Lotion.