UHPLC法同时测定苏子降气丸中10个成分含量的研究

目的：建立超高效液相色谱法（UHPLC法）同时测定苏子降气丸中阿魏酸、沉香四醇、甘草苷、橙皮苷、迷迭香酸、甘草酸铵、白花前胡甲素、和厚朴酚、白花前胡乙素、厚朴酚10个成分含量。方法：采用UHPLC方法，使用AQUITY UPLC BEH C₁₈色谱柱（2.1 mm×100 mm，1.8 μm），以乙腈-0.1%磷酸溶液为流动相，梯度洗脱，流速0.2 mL·min^-1、检测波长280 nm、柱温30℃、进样量2 μL。结果：阿魏酸、沉香四醇、甘草苷、橙皮苷、迷迭香酸、甘草酸铵、白花前胡甲素、厚朴酚、白花前胡乙素和厚朴酚含量测定结果分别为0.102 3~0.135 2、0.132 6~0.164 1、0.584 7~0.634 1、4.725~5.215、0.405 8~0.475 2、2.25 8~2.684、1.105~1.401、0.763 2~0.962 1、0.319 6~0.352 5、1.762~2.215 mg·g^-1；线性范围分别为1.001~18.02 μg·mL^-1（r=0.999 8）、1.224~22.04 μg·mL^-1（r=0.999 6）、4.778~86.00 μg·mL^-1（r=0.999 4）、38.80~698.4 μg·mL^-1（r=0.999 7）、3.752~67.54 μg·mL^-1（r=0.999 5）、20.52~369.4 μg·mL^-1（r=0.999 5）、9.916~178.5 μg·mL^-1（r=0.999 4）、7.262~130.7 μg·mL^-1（r=0.999 8）、2.624~47.23 μg·mL^-1（r=0.999 5）、14.86~267.5 μg·mL^-1（r=0.999 4）；定量下限分别为0.862、1.124、0.905、0.921、1.172、1.231、1.201、1.132、1.057、1.325 μg·mL^-1；平均加样回收率（n=6）分别为99.2%、98.9%、98.8%、98.6%、99.0%、98.6%、98.5%、98.8%、98.6%、98.4%（RSD<2.0%）；精密度（n=6）、重复性（n=6）、稳定性（48 h）（n=7）试验的RSD<2.0%。结论：所建立的多成分测定方法快捷、准确，重复性好，可用于苏子降气丸的质量控制。

Objective: To establish an UHPLC method for the simultaneous determination of 10 components in Suzi Jiangqi pills including ferulic acid,agarotetrol,liquiritin,hesperidin,rosmarinic acid,ammonium glycyrrhizinate,praeruptorin A,honokiol,praeruptorin B and magnolol. Methods: UHPLC was adopted using AQUITY UPLC BEH C₁₈(2.1 mm ×100 mm,1.8 μm)column and the mobile phase was acetonitrile-0.1% phosphoric acid solution with gradient elution at a flow rate of 0.2 mL·min^-1. The detection wavelength was 280 nm,and the column temperature was 30℃ and the injection volume was 2 μL. Results: Determination results of ferulic acid,agarotetrol,liquiritin,hesperidin,rosmarinic acid,ammonium glycyrrhizinate,praeruptorin A,honokiol,praeruptorin B and magnolol were 0.102 3-0.135 2 mg·g^-1,0.132 6-0.164 1 mg·g^-1,0.584 7-0.634 1 mg·g^-1,4.725-5.215 mg·g^-1,0.405 8-0.475 2 mg·g^-1,2.258-2.684 mg·g^-1,1.105-1.401 mg·g^-1,0.763 2-0.962 1 mg·g^-1,0.319 6-0.352 5 mg·g^-1 and 1.762~2.215 mg·g^-1,respectively. The linear ranges of 10 components were 1.001-18.02 μg·mL^-1(r=0.999 8),1.224-22.04 μg·mL^-1(r=0.999 6),4.778-86.00 μg·mL^-1(r=0.999 4),38.80-698.4 μg·mL^-1(r=0.999 7),3.752-67.54 μg·mL^-1(r=0.999 5),20.52-369.4 μg·mL^-1(r=0.999 5),9.916-178.5 μg·mL^-1(r=0.999 4),7.262-130.7 μg·mL^-1(r=0.999 8),2.624-47.23 μg·mL^-1(r=0.999 5)and 14.86-267.5 μg·mL^-1(r=0.999 4),respectively. Limits of quantitation were 0.862,1.124,0.905,0.921,1.172,1.231,1.201,1.132,1.057 and 1.325 μg·mL^-1,respectively. The average recoveries(n=6)were 99.2%,98.9%,98.8%,98.6%,99.0%,98.6%,98.5%.,98.8%,98.6% and 98.4% respectively with RSDs all below 2.0%. RSDs of precision(n=6),reproducibility(n=6)and stability tests(48 h)(n=7)were below 2.0%. Conclusion: The established multi-component method is fast,accurate and reproducible. It can be used for the quality control of Suzi Jiangqi pills.

[1] 中华人民共和国药典2015年版.一部[S].2015:930,265,185,191 ChP 2015.Vol 1[S].2015:930,265,185,191
[2] 谷丽华,林晨,吴弢,等.紫苏子药材质量标准研究[J].中国中药杂志,2010,35(16):2087 GU LH,LIN C,WU T,et al.Quality standard of Perillae Fructus[J].China J Chin Mater Med,2010,35(16):2087
[3] 马屏南.紫苏子镇咳、祛痰、平喘作用的药理研究[J].中南药学,2015,10(10):135 MA PN.Antitussive,expectorant and antiasthmatic effects of Perillae Fructus[J].Cent South Pharm,2015,10(10):135
[4] 王远,郭萍,李晖,等.RP-HPLC法同时测定紫苏子中α-亚麻酸和亚油酸的含量[J].药物分析杂志,2012,32(2):252 WANG Y,GUO P,LI H,et al.RP-HPLC simultaneous determination of α-linolenic acid and linoleic acid in Fructus Perillae[J].Chin J Pharm Anal,2012,32(2):252
[5] 张淑洁,钟凌云.厚朴化学成分及其现代药理研究进展[J].中药材,2013,36(5):838 ZHANG SY,ZHONG LY.Advance in chemical composition and modern pharmacology studies of Magnoliae Officinalis Cortex[J]. J Chin Med Mater,2013,36(5):838
[6] 罗燕燕,刘效栓,肖正国,等.HPLC双波长法同时测定甘草药渣提取物中甘草苷、异甘草苷、甘草素、异甘草素及甘草酸含量[J].中国中医药信息杂志,2017,24(12):64 LUO YY,LIU XS,XIAO ZG,et al.Determination of liquiritin,isoliquiritin,glycyrrhizin,isoliquiritigenin,glycyrrhizin,isoliquiritigenin and glycyrrhizic acid in licorice extract by HPLC dual wavelength spectrophotometry[J].Chin J Inf TCM,2017,24(12):64
[7] 陈方,向阳,朱立彬,等.当归药材质量标准提升研究[J].中国药师,2018,21(10):1861 CHEN F,XIANG Y,ZHU LS,et al.Quality standard improvement for Angelica sinensis[J].China Pharm,2018,21(10):1861
[8] 杨立军,高辉,王雪萍,等.苏子降气丸中厚朴酚与和厚朴酚含量测定[J].辽宁中医杂志,2008,35(4):589 YANG LJ,GAO H,WANG XP,et al.Determination of magnolol and honokiol in Suzi Jiangqi pills[J].Liaoning J Tradit Chin Med,2008,35(4):589
[9] 黎轶.HPLC法测定苏子降气丸中厚朴酚与和厚朴酚的含量[J]. 湖北中医杂志,2006,28(6):51 LI Y.Determination of magnolol and honokiol in Suzi Jiangqi pills by HPLC[J].Hubei J Tradit Chin Med,2006,28(6):51
[10] 张凤瑞.苏子降气丸质量标准研究[J].中成药,2005,27(7):775 ZHANG FR.Quality standard of Suzi Jiangqi pills[J].Chin Tradit Pat Med,2005,27(7):775
[11] 陈少萍,王丽丽,梁艺坚.超高效液相色谱法同时测定通宣理肺丸中白花前胡甲素和白花前胡乙素[J].中国中医药信息杂志,2018,25(9):76 CHEN SP,WANG LL,LIANG YJ.Simultaneous determination of praeruptorion A and praeruptorion B in Tongxuan Lifei pills by UPLC[J].Chin J Inf TCM,2018,25(9):76
[12] 陈亚,江滨,曾元儿.沉香色酮类化学成分研究[J].中国现代中药,2011,13(1):21 CHEN Y,JIANG B,ZENG YE.Study on chromones of Aquilariae Lignum Resinatum[J].Mod Chin Med,2011,13(1):21
[13] POIVRE M,DUEZ P.Biological activity and toxicity of the Chinese herb Magnolia officinalis Rehder & E.Wilson(Houpo)and its constituents[J].浙江大学学报B辑(生物医学与生物技术):英文版,2017,18(3):194
[14] 李欣然,王静,余晓燕,等.陈皮的研究进展[J].中国医药指南,2016,14(24):31 LI XR,WANG J,YU XY,et al.Advances of research work in Citri Reticulatae Pericarpium[J].Guide China Med,2016,14(24):31
[15] 李柯柯,任顺成.陈皮中黄酮类化合物的研究进展[J].食品研究与开发,2017,38(17):221 LI KK,REN SC.The development of flavonoids in dried orange peel[J].Food Res Dev,2017,38(17):221
[16] 杨爱霞,易凯,林世和.双波长高效液相色谱法同时测定祛白颗粒中二苯乙烯苷、阿魏酸及丹皮酚的含量[J].中国药师,2018,21(6):1093 YANG AX,YI K,LIN SH.Simultaneous determination of stilbene glycosides,ferulic acid and paeonol in Qubai granules by a dual-wavelength HPLC method[J].China Pharm,2018,21(6):1093
[17] 朱艳华,曹蓉蓉,张卫莲.生化丸质量标准的提高和优化[J].中国药师,2017,20(11):2082 ZHU YH,CAO RR,ZHANG WL.Improvement and optimization of the quality standard for Shenghua pills[J].China Pharm,2017,20(11):2082
[18] 王元,秦龙,李平,等.不同浓度乙醇对当归保肝活性成分超声提取的影响[J].中国药师,2016,19(8):1479 WANG Y,QIN L,LI P,et al.Effect of ethanol at different concentrations on liver protection active components in Angelica sinensis extracted by ultrasonic wave[J].Chin Pharma,2016,19(8):1479