HPLC法测定黄连、黄芩药对提取物中11个成分的含量

HPLC determination of 11 constituents in combination extracts of Coptidis Rhizoma and Scutellariae Radix

目的:建立高效液相色谱法测定黄连、黄芩药对提取物中11个成分(药根碱、表小檗碱、黄连碱、巴马汀、小檗碱、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、白杨素、千层纸素A)的含量。方法:采用Phenomoil 5μC18BDS(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-水(37∶63,内含磷酸二氢钾0.3400 g,十二烷基硫酸钠0.1700 g,磷酸调pH约为4),流速1.0 mL·min-1,检测波长349 nm,柱温30℃,对药对中黄连进行测定;采用Agilent ZORBAX SB-C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.3%磷酸三乙胺(B)水溶液为流动相,梯度洗脱,流速1.0 mL·min-1,检测波长为275 nm,柱温30℃,对药对中黄芩进行测定。结果:药根碱、表小檗碱、黄连碱、巴马汀、小檗碱、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、白杨素、千层纸素A进样量分别在0.015～0.15μg(r=0.9997)、0.033～0.33μg(r=0.9993)、0.17～1.7μg(r=0.9991)、0.079～0.79μg(r=0.9991)、0.31～3.1μg(r=0.9991)、0.42～4.2μg(r=0.9992)、0.078～0.78μg(r=0.9995)、0.053～0.53μg(r=0.9993)、0.016～0.16μg(r=0.9993)、1.2×10-3～0.012μg(r=0.9991)、0.010～0.10μg(r=0.9994)范围内呈良好线性关系;平均回收率(n=5)分别为100.1%、97.7%、97.6%、102.2%、99.4%、99.3%、99.9%、99.8%、101.1%、101.1%、100.3%,RSD分别为2.1%、1.1%、1.2%、0.9%、1.2%、1.8%、1.8%、2.2%、2.0%、1.9%、2.3%。结论:该方法为黄芩、黄连药对提取物的质量综合评价提供了科学依据。

Objective ：To establish an HPLC method for determination of eleven constituents （jatrorrhizine, epiber- berine, coptisine, palmatine, berberine, baicalin, wogonoside, baiealein, wogonin, chrysin, oroxylin- A） in combination extracts of the rhizome of Coptis chinensis Franch. and the root of Scutellaria baicalensis Georgi. Meth- ods：The Phenomoil 5μ C18 BDS column （250 mm ×4. 6 mm, 5 μm） with a column temperature of 30 ℃ was used ;the mobile phase consisted of aeetonitrile - water （ 37 ： 63, containing 0. 3400 g potassium phosphate and 0. 1700 g SDS, adjusted pH to 4） with flow rate of 1.0 mL. min-1 , and the detection wavelength was set at 349 nm for determination of above - mentioned alkaloids in the rhizome of C. chinensis. And the Agilent ZORBAX SB -C18 column（250 mm ×4.6 mm, 5 μm） with a column temperature of 30 ℃ was used;the mobile phase was acetonitrile（A） -0. 3% phosphoric acid -triethylamine （B） at gradient elution program and with a flow rate of 1.0 mL.mun -1, and the detection wavelength was set at 275 nm for determination of above - mentioned flavonoids in the root of S. baicalensis. Results： The content of eleven components in combination extracts of the rhizome of C. chinensis and the root of S. baicalensis was stable. The developed method had a good linearity in the ranges of 0. 015 -0. 15 μg（r =0. 9997） for jatrorrhizine, 0. 033 -0. 33 μg（r =0. 9993） for epiberberine, 0. 17 - 1.7 μg （r = 0. 9991 ） for coptisine, 0. 079 - 0. 79 μg（r = 0. 9991 ） μg for palmatine, 0.31 - 3.1μg（ r = 0. 9991 ） for ber-berine, 0. 42 -4.2 μg（r =0. 9992） for baicalin, 0. 078 -0.78 μg（r =0. 9995） for wogonoside, 0. 053 -0. 53 μg （r =0. 9993） for baicalein, 0. 016 -0. 16 μg（r =0. 9993） for wogonin, 1.2 x 10-3 -0. 012 μg（r =0. 9991 ） for ehrysin, 0. 010 -0. 10 μg（r =0. 9994） for oroxylin -A. The average reeoveries（n =5） were 100. 1%, 97.7%, 97.6%, 102.2%, 99.4%, 99.3%, 99.9%, 99.8%, 101.1%, 101.1%, 100.3% with RSDs of 2. 1%, 1.1%, 1.2%,0.9%, 1.2%, 1.8%, 1.8%, 2.2%, 2. 0%, 1.9%, 2. 3%, respectively. Conclusion： The developed method provides a scientific basis for quality assessment of combination extracts of the rhizome of C. chinensis and the root of S. baicalensis.