Tang Dan, Cao Dong-Min, Tan Lan-Fang, Xu You-Hua, Duan Ting-Ting, Zhu Quan, Wang Shu-Mei
School of Traditional Chinese Medicine, Guangdong Pharmaceutical University Guangzhou 510006, China Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of SATCM Guangzhou 510006, China Engineering Technology Research Center for Chinese Materia Medica Quality of Guangdong Province Guangzhou 510006, China Engineering & Technology Research Center for Chinese Materia Medica Quality of the Universities of Guangdong Province Guangzhou 510006, China.
State Key Laboratory of Quality Research in Chinese Medicine, Faculty of Chinese Medicine,Macau University of Science and Technology Macau 999078, China.
Zhongguo Zhong Yao Za Zhi. 2019 Apr;44(7):1410-1415. doi: 10.19540/j.cnki.cjcmm.20190118.006.
This research aims to develop an UHPLC method, based on core-shell column(i.e. superficially porous particles), for simultaneous determination of eight isoflavonoids including formononetin,(6αR,11αR)-3-hydroxy-9,10-dimethoxypterocarpan, calycosin-7-O-β-D-glucopyranoside,(3R)-7,2-dihydroxy-3,4-dimethoxyisoflavone, calycosin, ononin,(6αR,11αR)-9,10-dimethoxypterocarpan-3-O-β-D-glucopyranoside, and(3R)-7,2-dihydroxy-3,4-dimethoxyisoflavan-7-O-β-D-glucopyranoside in Astragali Radix. The analysis was performed on an Agilent Poroshell EC-C_(18 )column(2.1 mm×100 mm, 2.7 μm) with 0.2% formic acid solution(A)-acetonitrile(B) as mobile phase for gradient elution. The flow rate was 0.5 mL·min(-1), with column temperature of 40 ℃ and the wavelengths were set at 260 and 280 nm. According to the results, all calibration curves showed good linearity(R2>0.999 8) within the tested concentration ranges. Both the intra-and inter-day precisions for 8 isoflavonoids were less than 0.80%, with the mean recovery at the range of 94.71%-104.6%. Thus, the newly developed UHPLC method using core-shell column owned the advantages in terms of rapid analysis, low column pressure and less solvent consumption, thus enabling the usage of conventional HPLC systems. Meanwhile, quantitative evaluation was carried out for 22 batches of commercial Astragali Radix. It has been found that great variations occurred for the content of the individual isoflavonoids among different batches; in contrast, the total content of total 8 isoflavonoids(>0.1%) was stable in most samples, indicating that it was reasonable to involve all isoflavonoids as the chemical markers for the quality control of Astragali Radix.
本研究旨在开发一种基于核壳柱(即表面多孔颗粒)的超高效液相色谱法,用于同时测定黄芪中包括芒柄花黄素、(6αR,11αR)-3-羟基-9,10-二甲氧基紫檀烷、毛蕊异黄酮葡萄糖苷、(3R)-7,2-二羟基-3,4-二甲氧基异黄酮、毛蕊异黄酮、鹰嘴豆芽素A、(6αR,11αR)-9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷和(3R)-7,2-二羟基-3,4-二甲氧基异黄烷-7-O-β-D-葡萄糖苷在内的8种异黄酮类化合物。分析采用安捷伦Poroshell EC-C₁₈柱(2.1 mm×100 mm,2.7 μm),以0.2%甲酸溶液(A)-乙腈(B)为流动相进行梯度洗脱。流速为0.5 mL·min⁻¹,柱温为40℃,检测波长设定为260和280 nm。结果表明所有校准曲线在测试浓度范围内均呈现良好的线性关系(R²>0.999 8)。8种异黄酮类化合物的日内和日间精密度均小于0.80%,平均回收率在94.71% - 104.6%之间。因此,新开发的基于核壳柱的超高效液相色谱法具有分析速度快、柱压低和溶剂消耗少等优点,可使用常规高效液相色谱系统。同时,对22批市售黄芪进行了定量评价。结果发现,不同批次中各单种异黄酮类化合物的含量存在很大差异;相比之下,大多数样品中8种异黄酮类化合物的总含量(>0.1%)较为稳定,这表明将所有异黄酮类化合物作为黄芪质量控制的化学指标是合理的。