TLC结合HPLC-DAD法检查红花及西红花中13个红色色素

Test of thirteen red pigments in Carthami Flos and Stigma Croci by TLC and HPLC-DAD

目的:建立红花、西红花中13个非法添加红色色素的薄层色谱(TLC)和高效液相色谱(HPLC)检测方法。方法:采用TLC法,μL乙酸乙酯-正丁醇-乙醇-氨水-水(1∶3∶3∶1∶1);检测:日光下检视。采用HPLC-DAD梯度洗脱技术。色谱柱:Welch Ultimate^■XB-C18反相色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈-0.05 mol·L-1乙酸铵溶液为流动相流速:1.0 mL·min^-1,柱温35℃;检测波长:508 nm。并进行TLC和HPLC检测方法的检测下限、专属性及耐用性研究。通过比对对照品的比移值、保留时间与光谱图,确定红花样品与西红花样品中非法添加的色素。结果:已建立的13个红色色素的TLC与HPLC分析法灵敏度较高,专属性强,耐用性好。在TLC与HPLC佐证一致的情况下,22批红花药材中有10批检出胭脂红,有11批检出酸性红73;5批西红花药材中1批检出胭脂红、酸性红73、新品红、罗丹明B,1批检出胭脂红、新品红,1批检出胭脂红。结论:建立的TLC及HPLC检测方法经方法学验证,可用于红花、西红花药材中13个红色色素的检查。市售红花与西红花药材严重存在色素非法添加问题,应加大对中药材、中药饮片市场的监管力度。

Objective:To establish TLC and HPLC-DAD methods to detect the illegally added thirteen red pigments in Carthami Flos and Stigma Croci.Methods:TLC was used in the study.Ethyl acetate-butanolethanol-ammonia-water(1∶3∶3∶1∶1)was used as developing solvent and the plate was detected under visible light.HPLC-DAD with gradient elution was used in the research.The column was WelchUltimate?XB-C18(250 mm×4.6 mm,5μm)and the mobile phase was acetonitrile and 0.05 mol·L-1 ammonium acetate at a flow rate of 1 mL·min^-1.The detection wavelength was 508 nm and the column temperature was 35℃.Detection limit,specificity and durability of TLC and HPLC methods were evaluated.By comparing with the Rf values,retention time and spectra of the reference standards,the pigments in Carthami Flos and Stigma Croci were detected.Results:Based on consistent results from TLC and HPLC tests,10 and 11 out of 22 Carthami Flo samples were proven to contain ponceau 4 r and forbidden food pigment acid red 73,respectively.Each 1 out of 5 Stigma Croci samples were proven to contain ponceau 4 r,acid red 73,new fuchsin and rhodamine B,new fuchsin,and ponceau 4 r.Conclusion:The methods have been validated to be feasible for detection of thirteen red pigments in Carthami Flos and Stigma Croci.The results showe that illegally added pigments in Carthami Flos and Stigma Croci are severe.The government should strengthen the supervision on Chinese material medica and the prepared slides.