HPLC指纹图谱与一测多评法相结合的姜炭质量控制方法探索

Exploration of Combinational Quality Control Method of Carbonized Ginger Based on HPLC Fingerprint and QAMS

目的:建立姜炭饮片的HPLC指纹图谱,并应用一测多评法同时对该饮片中姜酮,6-姜辣素,6-姜烯酚,10-姜酚,8-姜烯酚,10-姜烯酚的含量进行测定。方法:采用Waters Symmetry ShieldTMRP18色谱柱(4.6 mm×250 mm,5μm),以乙腈-纯水为流动相进行梯度洗脱(0~30 min,25%~70%A;30~50 min,70%~90%A;50~60 min,90%A),流速1 m L·min^-1,检测波长240 nm,柱温30℃,以姜酮,6-姜辣素,8-姜酚,6-姜烯酚,10-姜酚,8-姜烯酚,10-姜烯酚为指标性成分建立姜炭饮片的指纹图谱。以6-姜辣素为参照内标,通过相对校正因子对姜酮,6-姜烯酚,10-姜酚,8-姜烯酚,10-姜烯酚进行含量测定,检测波长选择280 nm和220 nm。结果:建立了姜炭饮片HPLC指纹图谱,标定了10个共有峰,对7个共有峰进行了化学成分确认,分别为姜酮,6-姜辣素,8-姜酚,6-姜烯酚,10-姜酚,8-姜烯酚,10-姜烯酚。对姜炭中6个成分用一测多评法进行了测定,计算值与外标法实测值无明显差异。建议姜炭的质控指标为以干燥品计,姜酮(C11H14O3)不得少于0.020%,6-姜辣素(C17H26O4)不得少于0.050%,6-姜烯酚(C17H24O3)不得少于0.120%,10-姜酚(C21H34O4)不得少于0.080%,8-姜烯酚(C19H28O3)不得少于0.030%,10-姜烯酚(C21H32O3)不得少于0.050%。结论:所建立的方法准确、可行,为姜炭饮片的质量控制提供了简单有效的评价方法。

Objective:To establish the HPLC fingerprint of carbonized ginger and to determine the contents of zingerone,6-gingerol,6-shogaol,10-gingerol,8-shogaol and 10-shogaol with quantitative analysis of multi-components by single marker(QAMS).Method:The fingerprint of carbonized ginger was established by HPLC.All samples were analyzed by Waters SymmetryShieldTMRP18 column(4.6 mm×250 mm,5μm)with gradient elution by acetonitrile(A)-water(B)(0-30 min,25%-70%A;30-50 min,70%-90%A;50-60 min,90%A),the flow rate was 1.0 m L·min^-1,the detection wavelength was set at 240 nm and the column temperature was 30℃.Zingerone,6-gingerol,8-gingerol,6-shogaol,10-gingerol,8-shogaol and 10-shogaol was chosen as marker ingredients to establish HPLC fingerprint of carbonized ginger decoction pieces.Taking 6-gingerol as internal reference standard,the contents of zingerone,6-shogaol,10-gingerol,8-shogaol and 10-shogaol were determined at the detection wavelength of 220 nm and 280 nm according to the relative correction factor.Result:The HPLC fingerprint of carbonized ginger was obtained and 10 common peaks were designated,and 7 of them were identified as zingerone,6-gingerol,8-gingerol,6-shogaol,10-gingerol,8-shogaol and 10-shogaol,respectively.And there were no significant differences between the quantitative results of external standard method and QAMS.It is suggested that the content limits of carbonized ginger should be not less than 0.020%of zingerone(C11H14O3),0.050%of 6-gingerol(C17H26O4),0.120%of 6-shogaol(C17H24O3),0.080%of 10-gingerol(C21H34O4),0.030%of 8-shogaol(C19H28O3)and 0.050%of 10-shogaol(C21H32O3)calculated with reference to the dried products,respectively.Conclusion:The developed method is accurate and feasible,which can provide a simple and effective method for the quality control of carbonized ginger.