HPLC法测定杜仲-淫羊藿药对中8个化学成分的含量
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摘要
目的:建立HPLC分析方法同时测定杜仲-淫羊藿药对中绿原酸、咖啡酸、松脂醇二葡萄糖苷、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷Ⅰ8个成分的含量,明确配伍前后各成分含量的变化。方法:采用HPLC多波长切换-梯度洗脱技术。色谱柱为Eclipse XDB-C_(18)(4.6 mm×250 mm,5μm);流动相为乙腈(A)-0.1%磷酸水溶液(B);体积流量1.0 mL·min~(-1);柱温25℃;进样量5μL;检测波长:绿原酸、咖啡酸320 nm,松脂醇二葡萄糖苷277 nm,朝藿定A、朝藿定B、朝藿定C 270 nm,淫羊藿苷、宝藿苷Ⅰ280nm。结果:杜仲-淫羊藿药对中绿原酸、咖啡酸、松脂醇二葡萄糖苷、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷Ⅰ8个成分均实现良好分离,进样量分别在4.933×10~(-2)～1.579μg、2.554×10~(-3)～8.172×10~(-2)μg、3.957×10~(-2)～1.266μg、4.493×10~(-2)～1.438μg、5.979×10~(-2)～1.913μg、5.108×10~(-2)～1.635μg、0.188 0～6.016μg、1.701×10~(-2)～0.544 3μg范围内与峰面积均呈良好的线性关系(r>0.999);平均加样回收率为97.3%～103.8%(RSD<3%,n=6)。配伍后绿原酸、咖啡酸、淫羊藿苷、宝藿苷Ⅰ的含有量均有不同程度的减少,朝藿定B含有量有不同程度的增加,松脂醇二葡萄糖苷、朝藿定A、朝藿定C含有量无明显变化。结论:本方法快速、稳定、准确、简便,可用于杜仲-淫羊藿药对及其制剂的质量评价与质量控制,并为杜仲、淫羊藿的临床应用及后续配伍研究提供科学依据。
Objective:To establish an HPLC method for simultaneous determination of chlorogenic acid,caffeic acid,pinoresinol diglucoside,epimedin A,epimedin B,epimedin C,icariin and baohuoside Ⅰ in drug pair of Eucommiae Cortex and Epimedii Folium,so as to clear the variation trend in the contents of all ingredients after compatibility.Methods:HPLC multi-wavelength switching and gradient elution was used.Separation was performed on an Eclipse XDB-C_(18) column(4.6 mm×250 mm,5 μm)and the mobile phase consisted of acetonitrile(A)-0.1% phosphoric acid aqueous solution(B)at a flow rate of 1.0 mL·min~(-1).The column temperature was 25 ℃ and the injection volume was 5 μL.Detection wavelength was set at 320 nm for chlorogenic acid and caffeic acid,at 277 nm for pinoresinol diglucoside,epimedin A,at 270 nm for epimedin B and epimedin C,and at 280 nm for icariin and baohuoside Ⅰ.Results:Good separation and linearity(r>0.999)were obtained for the 8 ingredients in the range of 4.933×10~(-2)-1.579 μg、2.554×10~(-3)-8.172×10~(-2) μg、3.957×10~(-2)-1.266 μg、4.493×10~(-2)-1.438 μg、5.979×10~(-2)-1.913 μg、5.108×10~(-2)-1.635 μg、0.188 0-6.016 μg、1.701×10~(-2)-0.544 3μg.The average recovery rates were 97.3%-103.8%(RSD<3%,n=6).After compatibility,the contents of chlorogenic acid,caffeic acid,icariin and baohuoside Ⅰ reduced by different degrees.The contents of epimedin B increased by different degrees while there was no significant change in the contents of pinoresinol diglucoside,epimedin A and epimedin C.Conclusion:The method is rapid,stable,precise and simple,and can be used to control and evaluate the quality of Eucommiae Cortex and Epimedii Folium;and can provide scientific basis for clinical application and follow-up compatibility study of drug pair of Eucommiae Cortex and Epimedii Folium and their preparations.
Objective:To establish an HPLC method for simultaneous determination of chlorogenic acid,caffeic acid,pinoresinol diglucoside,epimedin A,epimedin B,epimedin C,icariin and baohuoside Ⅰ in drug pair of Eucommiae Cortex and Epimedii Folium,so as to clear the variation trend in the contents of all ingredients after compatibility.Methods:HPLC multi-wavelength switching and gradient elution was used.Separation was performed on an Eclipse XDB-C_(18) column(4.6 mm×250 mm,5 μm)and the mobile phase consisted of acetonitrile(A)-0.1% phosphoric acid aqueous solution(B)at a flow rate of 1.0 mL·min~(-1).The column temperature was 25 ℃ and the injection volume was 5 μL.Detection wavelength was set at 320 nm for chlorogenic acid and caffeic acid,at 277 nm for pinoresinol diglucoside,epimedin A,at 270 nm for epimedin B and epimedin C,and at 280 nm for icariin and baohuoside Ⅰ.Results:Good separation and linearity(r>0.999)were obtained for the 8 ingredients in the range of 4.933×10~(-2)-1.579 μg、2.554×10~(-3)-8.172×10~(-2) μg、3.957×10~(-2)-1.266 μg、4.493×10~(-2)-1.438 μg、5.979×10~(-2)-1.913 μg、5.108×10~(-2)-1.635 μg、0.188 0-6.016 μg、1.701×10~(-2)-0.544 3μg.The average recovery rates were 97.3%-103.8%(RSD<3%,n=6).After compatibility,the contents of chlorogenic acid,caffeic acid,icariin and baohuoside Ⅰ reduced by different degrees.The contents of epimedin B increased by different degrees while there was no significant change in the contents of pinoresinol diglucoside,epimedin A and epimedin C.Conclusion:The method is rapid,stable,precise and simple,and can be used to control and evaluate the quality of Eucommiae Cortex and Epimedii Folium;and can provide scientific basis for clinical application and follow-up compatibility study of drug pair of Eucommiae Cortex and Epimedii Folium and their preparations.
引文
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[7]张立超,李士春,云才,等.绿原酸通过Shp2激活Erk1/2促进大鼠成骨细胞增殖的实验研究[J].中成药,2014,36(4):694ZHANG LC,LI SC,YUN C,et al.Chlorogenic acid promotes rat osteoblast proliferation via Shp2 activation Erk1/2[J].Chin Tradit Pat Med,2014,36(4):694
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[9]陈克明.淫羊藿总黄酮的抗骨质疏松作用机制[J].中华中医药杂志,2017,32(12):5486CHEN KM.Anti-osteoporotic mechanism of total flavonoids from Herba Epimedii[J].Chin J Tradit Chin Med Pharm,2017,32(12):5486
[10]肖芳,黄勤挽,范润勇,等.HPLC法同时测定杜仲3个药用部位中8种成分[J].中成药,2016,38(8):1783XIAO F,HUANG QW,FANG RY,et al.Simultaneous determination of eight constituents in three medicinal parts of Eucommiae Cortex by HPLC[J].Chin Tradit Pat Med,2016,38(8):1783
[11]邢婧,任红,汪轩,等.HPLC法同时测定淫羊藿-川芎药对8种化学成分的含量[J].药物分析杂志,2015,35(6):961XING J,REN H,WANG X,et al.Simultaneous determination of eight ingredients in drugpair of Epimedii Folium and Chuanxiong Rhizoma by HPLC[J].Chin J Pharm Anal,2015,35(6):961
[12]杜寅筱,汪轩,张琳,等.UPLC法测定淫羊藿-威灵仙药对4种化学成分含量.[J].中华中医药杂志,2016,31(11):4782DU YX,WANG X,ZHANG L,et al.Determination of contents of4 chemical components in herb pair of epimedium herb Chinese clematis root by using UPLC method[J].Chin J Tradit Chin Med Pharm,2016,31(11):4782
[13]刘慧,张盛,刘仲华.HPLC法同时测定杜仲皮中京尼平苷酸、绿原酸、京尼平苷和松脂醇二葡萄糖苷[J].中草药,2012,43(8):1547LIU H,ZHANG S,LIU ZH.Simultaneous determination of geniposidic acid,chlorogenic acid,geniposide,and pinoresinol diglucoside in Eucommiae Cortex by HPLC[J].Chin Tradit Herb Drugs,2012,43(8):1547
[2]严红梅,张振海,孙娥,等.中药治疗骨质疏松症的研究进展[J].中草药,2014,45(8):1174YAN HM,ZHANG ZH,SUN E,et al.Research progress on treatment of osteoporosis using Chinese materia medica[J].Chin Tradit Herb Drugs,2014,45(8):1174
[3]袁航,曹树萍,陈抒云,等.淫羊藿的化学成分及质量控制研究进展[J].中草药,2014,45(24):3631YUAN H,CAO SP,CHEN SY,et al.Research progress on chemical constituents and quality control of Epimedii Folium[J].Chin Tradit Herb Drugs,2014,45(24):3631
[4]杜贵友,曹春雨.中药治疗骨质疏松症实验研究进展[J].中国中药杂志,2011,36(4):401DU GY,CAO CY.Experimental research progress of Chinese herbal medicine in the treatment of osteoporosis[J].China J Chin Mater Med,2011,36(4):401
[5]肖亚平,曾杰,焦琳娜,等.补肾中药对骨质疏松症的治疗及其信号通路调节作用的研究进展[J].中国中药杂志,2018,43(1):21XIAO YP,ZENG J,JIAO LN,et al.Review for treatment effect and signaling pathway regulation of kidney-tonifying traditional Chinese medicine on osteoporosis[J].China J Chin Mater Med,2018,43(1):21
[6]王娟娟,秦雪梅,高晓霞,等.杜仲化学成分、药理活性和质量控制现状研究进展[J].中草药,2017,48(15):3228WANG JJ,QIN XM,GAO XX,et al.Research progress on chemical compounds,pharmacological action,and quality status of Eucommiae Cortex[J].Chin Tradit Herb Drugs,2017,48(15):3228
[7]张立超,李士春,云才,等.绿原酸通过Shp2激活Erk1/2促进大鼠成骨细胞增殖的实验研究[J].中成药,2014,36(4):694ZHANG LC,LI SC,YUN C,et al.Chlorogenic acid promotes rat osteoblast proliferation via Shp2 activation Erk1/2[J].Chin Tradit Pat Med,2014,36(4):694
[8]石其福.咖啡酸对破骨细胞形成及分化的影响[D].石家庄:河北医科大学,2010SHI QF.Effects of Caffeic Acid on the Formation and Differentiation of Osteoclasts[D].Shijiazhuang:Hebei Medical University,2010
[9]陈克明.淫羊藿总黄酮的抗骨质疏松作用机制[J].中华中医药杂志,2017,32(12):5486CHEN KM.Anti-osteoporotic mechanism of total flavonoids from Herba Epimedii[J].Chin J Tradit Chin Med Pharm,2017,32(12):5486
[10]肖芳,黄勤挽,范润勇,等.HPLC法同时测定杜仲3个药用部位中8种成分[J].中成药,2016,38(8):1783XIAO F,HUANG QW,FANG RY,et al.Simultaneous determination of eight constituents in three medicinal parts of Eucommiae Cortex by HPLC[J].Chin Tradit Pat Med,2016,38(8):1783
[11]邢婧,任红,汪轩,等.HPLC法同时测定淫羊藿-川芎药对8种化学成分的含量[J].药物分析杂志,2015,35(6):961XING J,REN H,WANG X,et al.Simultaneous determination of eight ingredients in drugpair of Epimedii Folium and Chuanxiong Rhizoma by HPLC[J].Chin J Pharm Anal,2015,35(6):961
[12]杜寅筱,汪轩,张琳,等.UPLC法测定淫羊藿-威灵仙药对4种化学成分含量.[J].中华中医药杂志,2016,31(11):4782DU YX,WANG X,ZHANG L,et al.Determination of contents of4 chemical components in herb pair of epimedium herb Chinese clematis root by using UPLC method[J].Chin J Tradit Chin Med Pharm,2016,31(11):4782
[13]刘慧,张盛,刘仲华.HPLC法同时测定杜仲皮中京尼平苷酸、绿原酸、京尼平苷和松脂醇二葡萄糖苷[J].中草药,2012,43(8):1547LIU H,ZHANG S,LIU ZH.Simultaneous determination of geniposidic acid,chlorogenic acid,geniposide,and pinoresinol diglucoside in Eucommiae Cortex by HPLC[J].Chin Tradit Herb Drugs,2012,43(8):1547