基于液相色谱质谱联用的复方鱼腥草合剂化学成分分析及特征图谱研究

目的: 建立复方鱼腥草合剂LC-MS特征图谱的质量评价方法,并对特征图谱中的主要成分进行鉴别。方法: 采用Kromasil C₁₈色谱柱(4.6 mm×250 mm,5 μm),流动相A为0.1%甲酸溶液,流动相B为甲醇,梯度洗脱(0~15 min,20%B→30%B;15~30 min,30%B→50%B;30~40 min,50%B→65%B;40~45 min,65%B→70%B),流速1.0 mL·min^-1;质谱负离子模式检测。结果: 从复方鱼腥草合剂中检测到21个特征峰,通过对照品比对和文献参考,推断了其中的20个成分,包括(R,S)-告依春、3-咖啡酰奎尼酸/4-咖啡酰奎尼酸、绿原酸、马钱苷、7-O-ethyl sweroside、獐芽菜苷、异连翘酯苷、secoxyloganin、连翘酯苷A、4,5-二咖啡酰奎宁酸/3,5-二咖啡酰奎宁酸、金丝桃苷、异槲皮苷、槲皮苷、连翘苷、黄芩苷、baicalein-6-glucuronide、汉黄芩苷/baicalein 6-methylether-7-O-β-galactopyranuronoside;特征图谱方法验证结果表明精密度、重复性、稳定性良好。结论: 本文建立的方法可用于复方鱼腥草合剂的整体质量评价。

Objective: To develop a fingerprint method for quality assessment of compound Yuxingcao mistura by liquid chromatography coupled with mass spectrometry(LC-MS). Methods: Separation was performed using Kromasil C₁₈ column(4.6 mm×250 mm,5 μm).The mobile phase consisted of 0.1% formic acid aqueous solution(A) and methanol(B) using a gradient elution of 20%B→30%B at 0-15 min,30%B→50%B at 15-30 min,50%B→65%B at 30-40 min,65%B→70%B at 40-45 min.The detection was performed by mass spectrometry using negative mode by selected ion monitoring(SIM). Results: Twenty one characteristic peaks were detected from compound Yuxingcao mistura,and 20 peaks,including(R,S)-goitrin,3-caffeoylquinic acid/4-caffeoylquinic acid,chlorogenic acid,loganin,7-O-ethyl sweroside,sweroside,isoforsythiaside,secoxyloganin,forsythoside A,4,5-di-caffeoylquinic acid/3,5-di-caffeoylquinic acid,hyperoside,isoquercitrin,quercitrin,phillyrin,baicalin,baicalein-6-glucuronide,wogonoside/baicalein 6-methylether-7-O-β-galactopyranuronoside,were identified by comparison of the retention time and MS data with those of reference substances and reported in reference.The fingerprint method validation of compound Yuxingcao mistura showed good precision,reproducibility and stability. Conclusion: The developed LC-MS fingerprint method can be used for global assessment of the quality of compound Yuxingcao mistura.

[1] LIU YQ(刘艳清),WANG HW(汪洪武),YI KZ(易康柱).Simultaneous determination of chlorogenic acid and baicalin in compound Yuxingcao mistura by HPLC(HPLC同时测定复方鱼腥草合剂中绿原酸与黄芩苷含量)[J].Chin Tradit Pat Med(中成药),2008,30(6):附14
[2] ZHU XY(朱晓燕),CAI JZ(蔡进章),REN B(任斌).Determination of baicalin in compound Yuxingcao mistura by HPLC(高效液相色谱法测定复方鱼腥草合剂中黄芩苷的含量)[J].China Pharm(中国药业),2005,14(5):37
[3] JI YQ(戢艳琼),LI HL(李洪亮),ZHANG QF(张秋芳).Determination of chlorogenic acid and baicalin in compound Yuxingcao mixtures by HPLC(HPLC法测定复方鱼腥草合剂中绿原酸、黄芩苷的含量)[J].China Pharm(中国药师),2012,15(6):828
[4] ZHAO P(赵萍).Determination of baicalin in compound Yuxingcao mistura by HPLC(HPLC法测定复方鱼腥草合剂中黄芩苷的含量)[J].Chin Tradit Herb Drugs(中草药),2002,33(6):516
[5] LI ZH(李转花),HU SY(胡思玉),YU QB(余启波),et al.Compound Yuxingcao mixture studied by HPLC fingerprint(复方鱼腥草的HPLC指纹图谱研究)[J].Pharm Clin Res(药学与临床研究),2012,20(4):369
[6] ChP 2010.Vol Ⅰ(中国药典2010年版.一部)[S].2010:191
[7] XIA Y(夏远),LI DZ(李弟灶),PEI ZZ(裴振昭),et al.Review on the chemical constituents of the flower buds of Lonicera japonica(金银花化学成分的研究进展)[J].Mod Chin Med(中国现代中药),2012,14(4):26
[8] LI HJ(李会军),LI P(李萍).Chemical constituents in flower buds of Lonicera japonica Thunb.(忍冬花蕾的化学成分研究)[J].Chem Ind Forest Prod(林产化学与工业),2005,25(3):29
[9] ZHANG LY(张丽媛),LI YB(李遇伯),LI LX(李利新),et al.Chemical constituents of Lonicera japonica Thunb.by RRLC-Q-TOF/MS(RRLC-Q-TOF/MS分析金银花的化学成分)[J].Cent South Pharm(中南药学),2012,10(3):204
[10] LUO B(罗彬),ZHANG JZ(张进忠).Study on chemical constituents of extract of Forsythia suspense(连翘提取物化学成分研究)[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2013,19(2):143
[11] ZHOU JJ(周家驹),XIE GR(谢桂荣),YAN JX(严建新).Handbook of Chemical Constituents in Traditional Chinese Medicines(中药原植物化学成分手册)[M].Beijing(北京):Chemical Industry Press(化学工业出版社),2004:88
[12] HAN L(韩乐),CHEN QX(陈巧霞),LIU XH(刘训红),et al.Simultaneous determination of six flavonoids in Scutellariae Radix by MEKC-DAD(MEKC-DAD同时测定黄芩中6个黄酮类成分的含量)[J].Chin J Pharm Anal(药物分析杂志),2013,33(9):1525