波长切换HPLC法同时测定六神曲中9个成分的含量

目的：建立波长切换HPLC法同时测定六神曲中苦杏仁苷、青蒿素、香草酸、芦丁、金丝桃苷、阿魏酸、槲皮苷、槲皮素、山柰酚共9个成分的含量。方法：采用Agela Venusil XBP C₁₈（2）硅胶色谱柱（4.6mm×250 mm，5 μm），以乙腈为流动相A，0.08%磷酸水溶液为流动相B，梯度洗脱，流速为1 mL·min^-1，柱温为30℃，波长切换（0~15 min，46~62 min，210 nm，检测苦杏仁苷、青蒿素；15~24.9 min，27~46 min，258 nm，检测香草酸、芦丁、金丝桃苷、槲皮苷、槲皮素、山柰酚；24.9~27 min，218 nm，检测阿魏酸）。结果：在上述条件下，1次进样可同时测定9个成分。苦杏仁苷、青蒿素、香草酸、芦丁、金丝桃苷、阿魏酸、槲皮苷、槲皮素、山柰酚的进样量分别在31.95~639 μg（r=0.999 5），595~11 900 μg（r=0.999 4），17.82~356.32μg（r=0.999 9），17.27~345.44 μg（r=0.999 9），20.88~417.60 μg（r=0.999 9），75.6~1 512 μg（r=0.999 9），15.93~318.50 μg（r=0.999 9），8.7~174 μg（r=0.999 8），13.2~264 μg（r=0.999 9）范围内与峰面积呈良好线性关系；平均加样回收率（n=6）均高于95.0%，RSD均小于3.0%。10批样品中，上述9个成分含量测定结果依次为9.720~57.265、0~554.585、2.223~32.172、1.790~26.464、1.395~34.503、14.433~82.195、0.265~16.167、0.133~7.463、0.866~5.433 μg·g^-1。结论：该方法简便，测定结果准确，可用于六神曲的质量控制。

Objective: To establish a method of simultaneous determination of amygdalin, artemisinin, vanillic acid,rutin,hyperoside,ferulic acid,quercitrin,quercetin and kaempferol in Massa Medicata Fermentata by HPLC with wavelength switching. Methods: Agela Venusil XBP C₁₈ (2) (4.6 mm×250 mm, 5 μm) was chosen, and gradient elution was conducted with acetonitrile as mobile phase A and 0.08% phosphoric acid as mobile phase B. The flow rate was 1.0 mL·min^-1 and the column temperature was 30℃. The detection wavelength was set at 210 nm for Amygdalin and artemisinin during 0-15 min and 46-62 min,258 nm for vanillic acid,rutin,hyperoside,quercitrin, quercetin and kaempferol during 15-24.9 min and 27-46 min,218 nm for ferulic acid during 24.9-27 min. Results: The linear ranges of amygdalin,artemisinin,vanillic acid,rutin,hyperoside,ferulic acid,quercitrin,quercetin and kaempferol were 31.95-639 μg (r=0.999 5),595-11 900 μg (r=0.999 4),17.82-356.32 μg (r=0.999 9),17.27-345.44 μg (r=0.999 9),20.88-417.60 μg (r=0.999 9),75.6-1 512 μg (r=0.999 9),15.93-318.50 μg (r=0.999 9),8.7-174 μg (r=0.999 8) and 13.2-264 μg (r=0.999 9), respectively. The average recoveries (n=6) were above more than 95.0% with RSD less than 3.0%. The content ranges of ten batches of pilot samples were 9.720-57.265,0-554.585,2.223-32.172,1.790-26.464,1.395-34.503,14.433-82.195,0.265-16.167,0.133-7.463 and 0.866-5.433 μg·g^-1,respectively. Conclusion: The method is simple and accurate,and can be used for the quality control of Massa Medicata Fermentata.

[1] 中华人民共和国药典2015年版. 四部[S]. 2015:419 ChP 2015. Vol Ⅳ[S].2015:419
[2] 张鹏,陈冰,李宝银,等. 六神曲的质量调查与评价研究[J]. 北京中医药,2017, 36(7):658 ZHANG P, CHEN B, LI BY, et al. The quality investigation and evaluation of Massa Medicata Fermentata[J]. Beijing J Tradit Chin Med, 2017, 36(7):658
[3] 刘振启,刘杰. 道地"六神曲"的工艺与混乱品的鉴别[J]. 首都医药, 2009, 16(1):40 LIU ZQ, LIU J. The production process of chinese drugs of Massa Medicata Fermentata and the identification of chaos[J].Cap Med, 2009, 16(1):40
[4] 任巧玲,宋潇潇. 六神曲的质量情况分析[J]. 中国现代药物应用, 2010, 4(10):113 REN QL, SONG XX. Analysis of the quality of Massa Medicata Fermentata[J]. Chin J Mod Drug Appl,2010, 4(10):113
[5] 王彬辉,章文红,张晓芬,等. 苦杏仁苷提取工艺及药理作用研究新进展[J]. 中华中医药学刊, 2014, 32(2):381 WANG BH,ZHANG WH,ZHANG XF,et al. New research progress on extraction process and harmacological effects of amygdalin[J]. Chin Arch Tradit Chin Med,2014, 32(2):381
[6] 刘冲,刘荫贞,乐智勇,等. 高效液相色谱-蒸发光散射检测器法测定六神曲中青蒿素的含量[J]. 中南药学,2017,15(2):225 LIU C, LIU YZ, LE ZY, et al. HPLC-ELSD determination of artemisinin in Liushenqu[J]. Cent South Pharm,2017,15(2):225
[7] 陈静,陈铭阳,孙宇峰,等. 六神曲中辣蓼的本草考证[J]. 中国现代中药,2017, 19(1):116 CHEN J, CHEN MY, SUN YF, et al. Herbal textual of Laliao in Liushenqu[J]. Mod Chin Med,2017, 19(1):116
[8] 何立美,陈玉婷,刘洪梅,等. 高效液相色谱法同时测定水辣蓼提取物中6种黄酮成分的含量[J]. 广东农业科学,2014, 13:94 HE LM, CHEN YT, LIU HM, et al. Simultaneous determination of six kinds of flavonoid in extract of Polygonum hydropiper L. by HPLC[J]. Guangdong Agric Sci,2014, 13:94
[9] 刘娟秀,罗益远,刘训红,等. 不同采收期苍耳草中酚酸类及蒽醌类成分的动态积累分析[J]. 中草药, 2016, 47(7):1204 LIU JX, LUO YY, LIU XH, et al. Dynamic changes of phenolic acids and anthraquinones in aerial parts of Xanthium sibiricum from different collection periods[J].Chin Tradit Herb Drugs, 2016, 47(7):1204
[10] 付小梅,刘德鸿,孙艳朝,等. 蒙古苍耳草化学成分研究[J]. 中药材,2016, 39(6):1308 FU XM, LIU DH, SUN YC, et al. Chemical constituents from Xanthium mongolicum[J]. J Chin Med Mater, 2016, 39(6):1308
[11] 谢彦博,徐云,孙盛,等. HPLC测定六神曲中青蒿素、芦丁和槲皮素含量[J]. 中国酿造,2014, 33(10):140 XIE YB, XU Y, SUN S, et al. Determination of artemisinin, rutin and quercetin in Massa Medicata Fermentata by HPLC[J]. China Bre,2014, 33(10):140
[12] 王丽芳,高文远,徐鑫,等. 鲜干品组方六神曲发酵前后指标成分的LC-MS测定及指纹图谱分析[J]. 中国实验方剂学杂志, 2017,24(1):8 WANG LF,GAO WY,XU X,et al. Determination of index components and fingerprint about medicated leaven composed of fresh or dry medicines before and after fermentation by LC-MS[J]. Chin J Exp Tradit Med Form,2017,24(1):8
[13] 马维维,戚岑聪,张艳聪,等. 六神曲固态协同发酵工艺优化及物质动态变化研究[J]. 中华中医药学刊, 2017, 35(5):1290 MA WW, QI CC, ZHANG YC, et al. Optimization of medicated leaven Solid-state fermentation process and research of material change in the process[J]. Chin Arch Tradit Chin Med, 2017, 35(5):1290
[14] 练晶军. 六神曲质量特征及发酵变化研究[D]. 北京:北京中医药大学, 2011 LIAN JJ. Study on the Quality Characteristics and Fermentation Changes of Massa Medicata Fermentata[D]. Beijing:Beijing University of Chinese Medicine,2011
[15] 唐亮,谭淑珍,华坤鹏,等. RH-HPLC法测定青蒿素中脱氢青蒿素及9-表-青蒿素的含量[J]. 湖南文理学院学报(自然科学版), 2016, 28(1):24 TANG L,TAN SZ,HUA KP,et al.Determination of artemisitene and 9-epi-artemisinin in artemisinin raw material by RH-HPLC[J]. J Hunan Univ Arts Sci (Sci Technol),2016, 28(1):24