目的: 建立分别以其中任何1个结合型蒽醌为内参物均可用于测定大黄药材中8个结合型蒽醌的一测多评法(QAMS法)。方法: 采用高效液相色谱法,色谱柱为Symmetry C18(250 mm×4.6 mm,5 μm),以甲醇(A)-0.1%磷酸水(B)为流动相进行梯度洗脱,流速为1.0 mL·min-1,柱温为30 ℃,检测波长为280 nm。分别以芦荟大黄素-8-O-葡萄糖苷、大黄酸-8-O-葡萄糖苷、大黄素-1-O-葡萄糖苷、大黄酚-1-O-葡萄糖苷、大黄酚-8-O-葡萄糖苷、芦荟大黄素-3-羟甲基-O-葡萄糖苷、大黄素-8-O-葡萄糖苷、大黄素甲醚-8-O-葡萄糖苷为内参物,采用多点校正法,建立另外7个待测成分与内参物的相对校正因子(RCFs),计算待测成分与内参物色谱峰的相对保留时间(RRTs)。采用RCFs计算分别以其中1个成分为内参物时28批大黄药材中另外7个待测成分的含量,以差异百分比(PD)为参数,与外标法(ESM)测定结果进行比较,验证QAMS的准确性。结果: 分别以芦荟大黄素-8-O-葡萄糖苷、大黄酸-8-O-葡萄糖苷、大黄素-1-O-葡萄糖苷、大黄酚-1-O-葡萄糖苷、大黄酚-8-O-葡萄糖苷、芦荟大黄素-3-羟甲基-O-葡萄糖苷、大黄素-8-O-葡萄糖苷、大黄素甲醚-8-O-葡萄糖苷为内参物时,另外7个待测成分与内参物的RCFs分别为1.435、0.624、1.206、0.849、1.718、0.563、0.549,0.697、0.435、0.841、0.592、1.197、0.392、0.383,1.603、2.3、1.932、1.36、2.753、0.897、0.88,0.83、1.191、0.518、0.704、1.425、0.464、0.455,1.178、1.691、0.736、1.421、2.024、0.663、0.647,0.582、0.835、0.363、0.702、0.494、0.328、0.32,1.788、2.566、1.117、2.157、1.518、3.072、0.982和1.822、2.616、1.137、2.197、1.547、3.13、1.025,RRTs分别为1.207、1.47、1.68、1.719、1.763、1.768、2.015,0.829、1.218、1.393、1.425、1.461、1.466、1.671,0.681、0.821、1.143、1.169、1.199、1.203、1.371,0.596、0.719、0.876、1.023、1.049、1.053、1.2,0.583、0.703、0.856、0.977、1.026、1.029、1.173,0.568、0.685、0.835、0.953、0.975、1.003、1.143,0.566、0.683、0.832、0.95、0.972、0.997、1.139和0.497、0.599、0.73、0.834、0.853、0.875、0.878。8个成分含量的QAMS法计算结果的均值与ESM法测定结果之间的PD均＜3%。结论: 所建立的QAMS法准确、可行,可在实际工作中只有任何1个对照品情况下同时测定大黄药材中8个结合型蒽醌的含量。
Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) method for the determination of eight anthraquinone glycosides in Rhei Radix et Rhizoma when any one of them was chosen as the internal reference substance. Methods: HPLC method was developed on a Symmetry C18 (250 mm×4.6 mm, 5 μm) column with methanol (A)-0.1% phosphoric acid (B) as mobile phase for a gradient elution. The flow rate was set at 1.0 mL·min-1, the temperature maintained at 30 ℃, and the detection wavelength was set at 280 nm. Aloe emodin-8-O-glucoside, rhein-8-O-glucoside, emodin-1-O-glucoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucoside, aloe emodin-3-CH2-O-glucoside, emodin-8-O-glucoside, and physcion-8-O-glucoside was chosen as the internal reference substance, respectively, the relative correction factors (RCFs) of other seven components to be measured to the internal reference substances were established by multi-point correction method, and the relative retention time (RRTs) of the chromatographic peaks of components to be measured to the ones of the internal reference substances were calculated, respectively. The contents of other seven components to be measured in 28 batches of Rhei Radix et Rhizoma were calculated by RCFs when one of the eight components was used as the internal reference substance respectively, and the percentage difference (PD) was used as a parameter to compare the results with that of external standard method (ESM) to verify the accuracy of QAMS. Results: When aloe emodin-8-O-glucoside, rhein-8-O-glucoside, emodin-1-O-glucoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucoside, aloe emodin-3-CH2-O-glucoside, emodin-8-O-glucoside, and physcion-8-O-glucoside was used as internal reference substance respectively, the RCFs of the other seven components to be measured to the internal reference substances were (1.435, 0.624, 1.206, 0.849, 1.718, 0.563, 0.549), (0.697, 0.435, 0.841, 0.592, 1.197, 0.392, 0.383), (1.603, 2.3, 1.932, 1.36, 2.753, 0.897, 0.88), (0.83, 1.191, 0.518, 0.704, 1.425, 0.464, 0.455), (1.178, 1.691, 0.736, 1.421, 2.024, 0.663, 0.647), (0.582, 0.835, 0.363, 0.702, 0.494, 0.328, 0.32), (1.788, 2.566, 1.117, 2.157, 1.518, 3.072, 0.982) and (1.822, 2.616, 1.137, 2.197, 1.547, 3.13, 1.025), and the RRTs were (1.207, 1.47, 1.68, 1.719, 1.763, 1.768, 2.015), (0.829, 1.218, 1.393, 1.425, 1.461, 1.466, 1.671), (0.681, 0.821, 1.143, 1.169, 1.199, 1.203, 1.371), (0.596, 0.719, 0.876, 1.023, 1.049, 1.053, 1.2), (0.583, 0.703, 0.856, 0.977, 1.026, 1.029, 1.173), (0.568, 0.685, 0.835, 0.953, 0.975, 1.003, 1.143), (0.566, 0.683, 0.832, 0.95, 0.972, 0.997, 1.139) and (0.497, 0.599, 0.73, 0.834, 0.853, 0.875, 0.878), respectively. The PD between the mean values of the contents of eight components calculated by QAMS and the results determined by ESM were less than 3%. Conclusion: The established QAMS method is accurate and feasible, and can simultaneously determine eight anthraquinone glycosides in Rhei Radix et Rhizoma with only one reference substance in practical work.
[1] 涂栩秀,唐琼,赵映娟,等.同时采用一测多评法和外标法测定灯盏细辛注射液中2类物质的含量[J].药物分析杂志,2021,41(5):798
TU XX, TANG Q, ZHAO YJ, et al. Simultaneous determination two types of substances in Dengzhan Xixin injection by quantitative analysis of multie-components with single marker and the external standard method[J].Chin J Pharm Anal,2021, 41(5): 798
[2] DOU ZH, DAI Y, ZHOU YZ, et al. Quality evaluation of rhubarb based on qualitative analysis of the HPLC fingerprint and UFLC-Q-TOF-MS/MS combined with quantitative analysis of eight anthraquinone glycosides by QAMS[J].Biomed Chromatogr, 2021, 35(6): e5074
[3] ZHU CS, LI XP, ZHANG B, et al. Quantitative analysis of multi-components by single marker——a rational method for the internal quality of Chinese herbal medicine[J].Integr Med Res, 2017, 6(1): 1
[4] 万红才,徐作刚,段萍,等.一测多评法测定乌蕨中5个黄酮成分含量[J].药物分析杂志,2021,41(12):2211
WAN HC, XU ZG, DUAN P, et al. Determination of 5 flavonoids in Sphenomeris chinensis by quantitative analysis of multi-components by single marker[J].Chin J Pharm Anal,2021, 41(12): 2211
[5] 曹瑞,窦志华,倪丽丽,等. HPLC指纹图谱、Q-TOF/MS定性及多成分定量相结合的大黄饮片质量评价[J].中草药,2019,50(5): 1100
CAO R, DOU ZH, NI LL, et al. Quality evaluation of rhubarb pieces by HPLC fingerprint, Q-TOF/MS qualitative and multi-component quantitative analysis[J].Chin Tradit Herb Drugs, 2019, 50(5): 1100
[6] 戴莹,施凯,窦志华,等.大黄标准汤剂量值传递规律研究[J].中草药,2021, 52(10): 2938
DAI Y, SHI K, DOU ZH,et al. Study on law of quality value transmitting of Rhei Radix et Rhizoma standard decoction[J].Chin Tradit Herb Drugs, 2021, 52(10): 2938
[7] 陈艳琰,唐于平,陈嘉倩,等.大黄资源化学研究进展与利用策略[J].中草药, 2018, 49(21): 5170
CHEN YY, TANG YP, CHEN JQ, et al. Research progress and utilization strategy on resource chemistry of Rhei Radix et Rhizoma[J].Chin Tradit Herb Drugs, 2018, 49(21): 5170
[8] 李冬华,张平,张明童,等.全国大黄药材及其饮片评价性抽验结果分析与一测多评法研究[J].中国药事, 2018, 32(6): 775
LI DH, ZHANG P, ZHANG MT, et al. Analysis of the nationwide random inspection results of Rheum officinale and its pieces in QAMS[J].Chin Phar Aff, 2018, 32(6): 775
[9] 中华人民共和国药典2020年版. 一部[S].2020: 24
ChP 2020. Vol Ⅰ[S].2020: 24
[10] 窦志华,曹瑞,卞理,等.正交试验法优选大黄中蒽醌类成分提取工艺[J].中草药,2018, 49(14): 3279
DOU ZH, CAO R, BIAN L, et al. Optimization of extract process of anthraquinones from Rhei Radix et Rhizoma by orthogonal test[J].Chin Tradit Herb Drugs, 2018, 49(14): 3279
[11] 王玉,杨雪,夏鹏飞,等. 大黄化学成分、药理作用研究进展及质量标志物的预测分析[J].中草药, 2019, 50(19): 4821
WANG Y, YANG X, XIA PF, et al. Research progress on chemical composition and pharmacological effects of Rhei Radix et Rhizoma and predictive analysis on quality markers[J].Chin Tradit Herb Drugs, 2019, 50(19): 4821
[12] 毛春芳,施忠,罗琳,等. HPLC法同时测定大黄中芦荟大黄素等11种成分的量[J].中草药,2014, 45(16): 2400
MAO CF, SHI Z, LUO L, et al. Simultaneous determination of 11 constituents in Rhei Radix et Rhizoma by HPLC[J].Chin Tradit Herb Drugs, 2014, 45(16): 2400
[13] EP 9.0[S].2017: 1496
[14] WIANOWSKA D. Hydrolytical instability of hydroxyanthraquinone glycosides in pressurized liquid extraction[J].Anal Bioanal Chem, 2014, 406(13): 3219
[15] CHEN AZ, SUN L, YUAN H, et al. Simultaneous qualitative and quantitative analysis of 11 active compounds in rhubarb using two reference substances by UHPLC[J].J Sep Sci, 2018, 41(19): 3686
[16] 胡樱凡,向丽,王平,等. 大黄肝肾毒性及其减毒方法现代研究进展[J].中国实验方剂学杂志, 2019, 25(11): 34
HU YF, XIANG L, WANG P, et al. Hepatotoxicity and nephrotoxicity of Rhei Radix et Rhizoma and its attenuation methods[J].Chin J Exp Med Form, 2019, 25(11): 34
[17] 邓诺.大黄及其总蒽醌肝肾毒性研究[D].北京:中国医学科学院, 2018
DENG N. Study on Liver and Kidney Toxicity of Rhubarb and Its Total Anthraquinones[D]. Beijing: China Academy of Chinese Medical Sciences, 2018
[18] TAN P, ZHANG L, ZHAO YL, et al. A practical method for the simultaneous quantitative determination of twelve anthraquinone derivatives in rhubarb by a single-marker based on ultra-performance liquid chromatography and chemometric analysis[J].Anal Methods, 2016, 8: 3927
[19] 刘移兰,蔡卫华,窦志华,等.一测多评法同时测定五味子中12个木脂素类成分的含量[J].药物分析杂志, 2019, 39(7):1207
LIU YL, CAI WH, DOU ZH, et al. Simultaneous determination of 12 lignans in Schisandrae Chinensis Fructus by quantitative analysis of multi-components with single marker[J].Chin J Pharm Anal,2019, 39(7): 1207
[20] 何兵,杨世艳,张燕.一测多评中待测成分校正和定位的新方法研究[J].药学学报, 2012, 47(12): 1653
HE B, YANG SY, ZHANG Y. A new method of calibration and positioning in quantitative analysis of multi components by single marker[J].Acta Pharm Sin, 2012, 47(12): 1653
[21] 石莉尧,房蕴歌,陈两绵,等.以蟾酥制剂为例,探讨一测多评法在中成药质量评价和控制中的应用研究[J].中国中药杂志, 2021, 46(12):2931
SHI LY, FANG YG, CHEN LM, et al. Application of QAMS for quality evaluation and control of Chinese patent medicines: taking Bufonis Venenum-contained preparations as examples[J].China J Chin Mater Med, 2021, 46(12): 2931
