UHPLC-FLD法测定不同产地大黄中6种大黄蒽醌

Determination of six rhubarb anthraquinones of Rhei Radix et Rhizoma decoction pieces from different growing areas by UHPLC-FLD

目的 建立超高效液相色谱荧光(UHPLC-FLD)法同时测定大黄中大黄素、芦荟大黄素、大黄酸、大黄酚、大黄素甲醚和大黄素-8-O-β-D-葡萄糖苷的含量,并应用于10个不同产地大黄饮片的分析。方法 大黄粉末经甲醇提取制备供试品溶液,采用Agilent Poroshell 120 EC-C_(18)柱(50 mm×4.6 mm,2.7μm);流动相为0.1%甲酸水溶液(A)-0.1%甲酸甲醇(B),梯度洗脱;体积流量0.6 m L·min^(-1);柱温30℃;荧光检测器激发波长435 nm,发射波长515 nm。进行专属性、线性关系、定量限及检出限、精密度、稳定性、重复性、加样回收率考察。应用已建立的方法 分析10个不同产地大黄饮片中6种大黄蒽醌含量。结果 6种成分在各自范围内线性关系良好,精密度、稳定性、重复性均良好;平均加样回收率为97.86%~103.2%,RSD为1.27%~2.15%。10个不同产地大黄饮片中蒽醌总量均符合《中国药典》要求;但不同产地的大黄中蒽醌单体的含量差别较大。结论 建立的UHPLC-FLD法灵敏、快速、准确、稳定且重复性好,可用于大黄中6种大黄蒽醌的含量测定。

Objective To establish a UHPLC-FLD method for the simultaneous determination of emodin,aloe emodin,rhein,chrysophanic acid,emodin-3-methyl ether and emodin-8-O-β-D-glucoside in Rhei Radix et Rhizom decoction pieces.Methods The analysis of methanol extract of Rhei Radix et Rhizom decoction piece was performed on a 30℃thermostatic Agilent Poroshell 120EC-C_(18)column (50 mm×4.6 mm,2.7μm),with the mobile phase comprising of 0.1% formic acid water (A)-0.1%formic acid methanol (B) flowing at 0.6 m L·min^(-1)in a gradient elution manner,and the excitation wavelength and emission wavelength were set at 435 nm and 515 nm,respectively.Specificity,linear relationship,limit of quantitation and detection,precision,stability,repeatability and recovery were investigated.The established method was used to analyze the anthraquinone content of six kinds of rhubarb from 10 different areas.The established method was used to analyze the anthraquinone content of six kinds of rhubarb from10 different areas.Results Six constituents showed good linear relationships within their own ranges.Precision,stability and repeatability all meet the testing requirements.whose average recoveries were 97.86%-103.2%with the RSDs of 1.27%-2.15%.The total amount of anthraquinone in 10 Rhei Radix et Rhizoma decoction pieces from different origins met the requirements of pharmacopoeia.However,the content of anthraquinone monomer in Rhei Radix et Rhizoma decoction pieces from different producing areas varies greatly.Conclusion The method is sensitive,rapid,accurate,stable and reproducible,and can be used for the determination of related compounds in Rhei Radix et Rhizom.