闪蒸-气相色谱/质谱法同时测定四神丸中8个有效成分

目的：建立闪蒸-气相色谱/质谱法（FE-GC/MS）同时测定四神丸中补骨脂素、异补骨脂素、五味子甲素、去氢二异丁香酚、五味子乙素、五味子醇甲、吴茱萸碱和吴茱萸次碱8个有效成分的含量。方法：将四神丸粉末样品置于裂解器中，在300℃下闪蒸后采用GC或GC/MS分析，色谱柱为UA-5（30 m×0.25mm×0.25 μm），载气为氦气，程序升温（初始温度50℃，以10℃·min^-1升至200℃，再以5℃·min^-1升至300℃，10 min），进样口温度300℃，FID（300℃）或质谱检测器（EI离子源，无溶剂延迟，扫描范围m/z 50~600）。结果：8个有效成分达到完全分离，重复性良好，RSD小于6.2%。8个成分在20~10 000 ng范围内线性关系良好，r>0.992，样品加标回收率为92.6%~107.5%。对7批四神丸样品进行了定量分析，并与HPLC法所得结果进行比较，通过五味子醇甲的含量变化及对安五脂素的质谱定性，可鉴别五味子的疑似掺杂。结论：该方法前处理简单，检测灵敏度高，准确可靠，可用于四神丸的质量控制。

Objective: To develop a flash evaporation-gas chromatography/mass spectrometry (FE-GC/MS) method for simultaneous determination of psoralen,isopsoralen,deoxyschizandrin,dehydrodiisoeugenol,γ-Schisandrin, schisandrin, evodiamine and rutaecarpine in Sishen pills. Methods: Powdered Sishen pills was directly added to a sample cup,then evaporated at 300℃ and the analytes were analyzed by GC or GC/MS. UA-5 capillary column (30 m×0.25 mm×0.25 μm) was used with temperature programming. The oven temperature was raised from 50℃ to 200℃ at a rate of 10℃·min^-1,then raised to 300℃ at a rate of 5℃·min^-1 and was maintained for 10 min. The injector was set at 300℃. FID (temperature was 300℃) or mass spectrometry (EI ion source with its scanning at the range of m/z 50-600, and no solvent delay). Results: The 8 components were well separated, and RSD was less than 5.3%,showing good reproducibility. The calibration curves exhibited good linearity with regression coefficients more than 0.992 at the conc entration range from 20 to 10 000 ng for all the components. The recoveries were 92.6%-107.5%. Finally, the contents of these 8 components in Sishen pills from 7 manufactories samples were analyzed, and the results were compared with those obtained by HPLC method. The Doping of Wuweizi might be detected by the content variation of schisandrin and qualitative analysis of anwuligan. Conclusion: The method was simple,sensitive, accurate,reliable,and could be used for quality control of Sishen pills.

[1] 谭生建,胡文祥,杨成勇,等.梯度洗脱变换波长HPLC测定四神丸中有效成分[J].中国药学杂志,1999, 34(12):839 TAN SJ, HU WX, YANG CY, et al. Gradient eluation with changeable-wavelength for determination of psoralen,isopsoralen, evodiamine and rrutaecarpine in Sishen Wan by HPLC[J]. Chin Pharm J,1999, 34(12):839
[2] 蔺晓源,邓娜,李霞,等.四神丸及其拆方对脾肾阳虚泄泻大鼠Leptin、CCK、IP3的影响[J].中医药导报,2017,23(3):22 LIN XY, DENG N, LI X, et al. Effect of the Sishenwan and its disassembled prescriptions on Leptin, CCK and IP3 of rats with diarrhea of asdthenic splenonephro-yang[J]. J Chin Med Pharmacol,2017, 23(3):22
[3] 中华人民共和国药典2015版.一部[S]. 2015:66 ChP 2015. Vol I[S]. 2015:66
[4] 刘欢,陈剑锋,王静,等.高效液相色谱法同时测定肉豆蔻麸煨前后肉豆蔻木脂素和去氢二异丁香酚的含量[J].医药导报, 2014, 33(8):1070 LIU H, CHEN JF, WANG J, et al. Determination of content of nutmeg lignan and dehydrodiisoeugenol in nutmeg by HPLC[J]. Her Med,2014, 33(8):1070
[5] 楼之岑,秦波.常用中药材品种整理和质量研究[M].第二卷.北京:北京医科大学中国协和大学联合出版社, 1995:299 LOU ZC,QIN B. Species Systematization and Quality Evaluation of Commonly Used Chinese Traditional Drugs[M]. Vol 2nd. Beijing:Peking Union Medical College and Beijing Medical University Press, 1995:299
[6] 王晓霞,高慧媛,姜勇,等.吴茱萸化学成分研究[J].中草药, 2013, 44(10):1241 WANG XX, GAO HY, JIANG Y, et al. Chemical constituents from fruits of Euodia rutaecarpa[J]. Chin Tradit Herb Drugs, 2013, 44(10):1241
[7] 姚兰,王运军,刘晓英,等. RP-HPLC法同时测定四神丸中补骨脂素、异补骨脂素、五味子醇甲和吴茱萸碱的含量[J].中国药房, 2010, 21(43):4095 YAO L, WANG YJ, LIU XY, et al. Simultaneous determination of psoralen,isopsoralen,schisandrin and evodiamine in Sishen pills by RP-HPLC[J]. Chin Pharm,2010, 21(43):4095
[8] 苏建,刘永利,郭毅,等.多波长HPLC法同时测定四神丸中7个有效成分的含量[J].药物分析杂志, 2013, 33(9):1531 SU J, LIU YL, GUO Y, et al. Simultaneous determination of seven active components in Sishen pills by HPLC under multiple UV wavelengths[J]. Chin J Pharm Anal,2013,33(9):1531
[9] 陈耀祖,李兆林,薛敦渊,等.闪蒸-毛细管气相色谱-质谱法分析中草药的挥发性成分[J].高等学校化学学报, 1985, 6(12):1075 CHEN YZ, LI ZL, XUE DY, et al. Analysis of volatile constituents of Chinese medicinal herbs by flash distillation-capillary gas chromatography-mass spectrometry[J]. Chem J Chin Univ, 1985, 6(12):1075
[10] LIANG MM, QI ML, ZHANG CB, et al. Gas chromatographymass spectrometry analysis of volatile compounds from Houttuynia cordata Thunb after extraction by solid-phase microextraction,flash evaporation and steam distillation[J]. Anal Chim Acta, 2005, 531(1):97
[11] 张聪,齐美玲,傅若农.闪蒸气相色谱质谱法测定中药川芎挥发性成分[J].世界科学技术-中医药现代化,2009, 11(1):165 ZHANG C,QI ML,FU RN. Flash evaporation-gas chromatographymass spectrometry for the analysis of volatile compounds in Ligusticum chuanxiong[J]. World Sci TechnolMod Tradit Chin Med, 2009, 11(1):165
[12] 祝璇,徐国钩,金蓉鸾,等.闪蒸一毛细管气相色谱一质谱法鉴定中药木香类的成分[J].中国药科大学学报, 1990(3):159 ZHU X, XU GG, JIN RL, et al. Study on Chinese traditional drug "Muxiang" by flash distillation-capillary-GC-MS[J]. J China Pharm Univ,1990(3):159
[13] 刘会君,陈爽,王鹏,等.莪术药材的闪蒸气相色谱测定及其模式识别分析研究[J].化学学报, 2012, 70(1):78 LIU HJ, CHEN S, WANG P, et al. Flash evaporation-gas chromatography determination with pattern recognition analysis for Curcuma zedoaria Rosc[J]. Acta Chim Sin, 2012, 70(1):78
[14] HUANG ZP, HUANG YL, XU SQ, et al. Discrimination of the traditional Chinese medicine from Schisandra Fruits by flash evaporation-gas chromatography/mass spectrometry and fingerprint analysis[J]. Chromatographia,2015, 78(15):1083
[15] 杜小弟,李俊生,郭丽萍,等.分散液液微萃取-气相色谱法快速测定水中15种硝基苯类物质[J]分析化学,2017, 45(11):1711 DU XD, LI JS, GUO LP, et al. Determination of 15 kinds of nitroaromatics in aqueous samples using dispersive liquid-liquid microextraction and gas chromatography with electron capture detection[J]. Chin J Anal Chem,2017, 45(11):1711
[16] 马虹英. GC法测定补骨脂酊中补骨脂素和异补骨脂素的含量[J].中国药房, 2010, 21(23):2170 MA YH. Content determination of psoralen and isopsoralen in Buguzhi Tincture by GC[J]China Pharm,2010, 21(23):2170