一测多评法同时测定五味子中12个木脂素类成分的含量

目的：建立可用于测定五味子中12个木脂素类成分的一测多评法（QAMS法）。方法：采用高效液相色谱法，色谱柱为Symmetry C₁₈（4.6 mm×250 mm，5 μm），以乙腈（A）-0.1%磷酸水（B）为流动相进行梯度洗脱（0~25 min，45% A→50% A；25~26 min，50% A→60% A；26~36 min，60% A→55% A；36~50 min，55% A→50% A；50~55 min，50% A→65% A；55~70 min，65% A；70~75 min，65% A→70% A；75~85 min，70% A），流速1.0 mL·min^-1，柱温30℃，检测波长217 nm。以五味子醇甲为内参物，采用多点校正法，分别建立戈米辛D、戈米辛J、五味子醇乙、当归酰戈米辛H、戈米辛G、五味子酯甲、五味子酯乙、五味子酚、五味子甲素、五味子乙素、五味子丙素11个待测成分与五味子醇甲的相对校正因子，计算待测成分与五味子醇甲色谱峰的相对保留值。采用相对校正因子计算14批五味子样品中11个待测成分的含量，并以差异百分比（PD）为参数，与外标法（ESM）测定结果进行比较，验证QAMS的准确性。另取9批五味子样品，使用3根色谱柱分别在2个实验室的3台仪器采用所建立的方法进行五味子醇甲的含量测定和11个待测成分的含量计算，比较测定和计算结果的相对标准偏差（RSD），验证所建立方法的实用性。结果：上述11个待测成分的RCF分别为1.127、0.883、0.679、0.772、0.712、0.483、0.736、0.451、0.404、0.767和0.754，相对保留值分别为1.179、1.263、1.342、1.810、2.025、2.230、2.280、2.448、3.529、3.996和4.303，QAMS法计算的11个待测木脂素类成分含量与ESM测定结果之间的PD均小于5%。另外9批样品的五味子醇甲含量测定结果和11个待测成分含量计算结果的RSD也均小于5%。23批五味子中上述11个待测成分及五味子醇甲的含量范围分别为0.173~0.395、0.197~0.438、0.571~1.124、0.518~1.016、0.105~0.214、0.076~0.715、0.399~0.812、0.061~0.135、0.349~1.250、1.308~2.376、0.126~0.444和3.571~6.764 mg·g^-1。结论：建立的QAMS法可实际应用于五味子中12个木脂素类成分含量测定。

Objective:To establish a quantitative analysis of multi-components by single marker(QAMS)method for the determination of 12 lignans in Schisandrae Chinensis Fructus.Methods:A high-performance liquid chromatography(HPLC)method was developed on a Symmetry C₁₈(4.6 mm×250 mm,5 μm)column with acetonitrile(A)-0.1% phosphoric acid(B)as mobile phase with gradient elution(0-25 min,45%A→50%A;25-26 min,50%A→60%A;26-36 min,60%A→55%A;36-50 min,55%A→50%A;50-55 min,50%A→65%A;55-70 min,65%A;70-75 min,65%A→70%A;75-85 min,70%A).The flow rate was set at 1.0 mL·min^-1,the temperature maintained at 30℃,and the detection wavelength was set at 217 nm.Schisandrol A was chosen as the internal reference substance,the relative correction factors of 11 components were determined by multi-point correction method,including gomisin D,gomisin J,schisandrol B,angeloylgomisin H,gomisin G,schisantherin A,schisantherin B,schisanhenol,deoxyschisandrin,schisandrin B and schisandrin C.And the relative retention values of the chromatographic peaks of the components to schisandrol A were calculated,respectively.The contents of 11 components in 14 batches of Schisandrae Chinensis Fructus were determined by relative correction factors,and the percentage difference(PD)was used as a parameter to compare the results with that of external standard method(ESM)to verify the accuracy of QAMS.Three chromatographic columns and three instruments were used in two laboratories to determine the content of schisandrol A,and to calculate the contents of 11 components in another nine batches of samples.And the feasibility of the established method was verified by the relative standard deviation(RSD)of the measured and calculated results.Results:The relative correction factors of 11 components were 1.127,0.883,0.679,0.772,0.712,0.483,0.736,0.451,0.404,0.767 and 0.754,respectively and the relative retention values were 1.179,1.263,1.342,1.810,2.025,2.230,2.280,2.448,3.529,3.996,and 4.303,respectively.The PDs of the contents of 11 lignans calculated by QAMS and determined by ESM were less than 5%.The RSDs of the measured content of schisandrol A and the calculated contents of 11 components in other nine batches of Schisandrae Chinensis Fructus were also less than 5%.The contents of 11 components and schisandrol A in 23 batches of Schisandrae Chinensis Fructus ranged from 0.173 to 0.395,0.197 to 0.438,0.571 to 1.124,0.518 to 1.016,0.105 to 0.214,0.076 to 0.715,0.399 to 0.812,0.061 to 0.135,0.349 to 1.250,1.308 to 2.376,0.126 to 0.444 and 3.571 to 6.764 mg·g^-1,respectively.Conclusion:The established QAMS method can be applied in the determination of 12 lignans in Schisandrae Chinensis Fructus.

[1] ZHU C,LI X,ZHANG B,et al.Quantitative analysis of multi-components by single marker-a rational method for the internal quality of Chinese herbal medicine[J].Integr Med Res,2017,6(1):1
[2] 秦昆明,杨冰,胡静,等.一测多评法在中药多组分质量控制中的应用现状与思考[J].中草药,2018,49(3):725 QIN KM,YANG B,HU J,et al.Application status and thinking of quantitative analysis of multi-components by single marker in quality control of multi-components traditional Chinese medicine[J].Chin Tradit Herb Drugs,2018,49(3):725
[3] DONG Y,GUO Q,LIU J,et al.Simultaneous determination of seven phenylethanoid glycosides in Cistanches Herba by a single marker using a new calculation of relative correction factor[J].J Sep Sci,2018,41(9):1913
[4] 杨洋,黄良永,朱美玲,等.一测多评法在中国药典2015年版中的应用[J].中南药学,2017,15(2):1738 YANG Y,HUANG LY,ZHU ML,et al.Quantitative analysis of multi-components with single-marker in China Pharmacopoeia(2015 edition)[J].Cent South Pharm,2017,15(2):1738
[5] 陈舒妤,石婧婧,邹立思,等.UFLC-Q-TRAP-MS/MS同时测定五味子中木脂素及有机酸类成分[J].中国中药杂志,2018,43(10):2104 CHEN SY,SHI JJ,ZOU LS,et al.Simultaneous determination of lignans and organic acids in Schisandrae Chinensis Fructus by UFLC-Q-TRAP-MS/MS[J].China J Chin Mater Med,2018,43(10):2104
[6] 安莉萍.五味子质量控制及血清药化学研究[D].南京:南京中医药大学,2014 AN LP.Study on the Quality Control and Serum Pharmacochemistry of Schisandra chinensis[D].Nnanjing:Nanjing University of Chinese Medicine,2014
[7] 窦志华,安莉萍,陈敏,等.一测多评法同时测定五味子中6种木脂素类成分含量[J].中国药学杂志,2014,49(2):147 DOU ZH,AN LP,CHEN M,et al.Simultaneous determination of six lignanoids in Schisandra chinensis by quantitative analysis of multi-components by single marker[J].Chin Pharm J,2014,49(2):147
[8] 贺凤成,李守信,赵志全,等.一测多评法测定五味子中4种木脂素类成分的含量[J].药学学报,2012,47(7):930 HE FC,LI SX,ZHAO ZQ,et al.Simultaneous quantitative analysis of four lignanoids in Schisandra chinensis by quantitative analysis of multi-components by single marker[J].Acta Pharm Sin,2012,47(7):930
[9] 汪丽君,刘淑敏,毛春芹,等.一测多评法同时测定五味子中8个木脂素类成分的含量[J].药物分析杂志,2015,35(7):1191 WANG LJ,LIU SM,MAO CQ,et al.Simultaneous determination of 8 lignans in Schisandra chinensis by quantitative analysis of multi-components by single marker[J].Chin J Pharm Anal,2015,35(7):1191
[10] ZHANG Y,CHEN Z,XU X,et al.Rapid separation and simultaneous quantitative determination of 13 constituents in Psoraleae Fructus by a single marker using high-performance liquid chromatography with diode array detection[J].J Sep Sci,2018,40(21):4191
[11] 中华人民共和国药典2015年版.四部[S].2015:375 ChP 2015.Vol Ⅳ[S].2015:375
[12] 王智民,钱忠直,张启伟,等.一测多评法建立的技术指南[J].中国中药杂志,2011,36(6):657 WANG ZM,QIAN ZZ,ZHANG QW,et al.The guidelines for establishment of QAMS[J].China J Chin Mater Med,2011,36(6):657
[13] 何兵,杨世艳,张燕.一测多评中待测成分校正和定位的新方法研究[J].药学学报,2012,47(12):1653 HE B,YANG SY,ZHANG Y,et al.A new method of calibration and positioning in quantitative analysis of multicomponents by single marker[J].Acta Pharm Sin,2012,47(12):1653