Wang Ting-Ge, Luo Lu-Yao, Xue Rui, Li Jin-Cao, Li Shuo, Zhang Yang, Zhang You-Zhi
Nanjing University of Chinese Medicine Nanjing 210023, China State Key Laboratory of Toxicology and Medical Countermeasures, Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences, Academy of Military Sciences Beijing 100850, China.
State Key Laboratory of Toxicology and Medical Countermeasures, Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences, Academy of Military Sciences Beijing 100850, China School of Pharmacy, North China University of Science and Technology Tangshan 063210, China.
Zhongguo Zhong Yao Za Zhi. 2024 Jan;49(1):141-150. doi: 10.19540/j.cnki.cjcmm.20230919.201.
This study established an HPLC fingerprint and multi-component content determination method for salt-fired Eucommiae Cortex, and evaluated the quality of salt-fired Eucommiae Cortex from different sources using fingerprint similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and orthogonal partial least square discriminate analysis(OPLS-DA). HPLC was launched on a Cosmosil 5C_(18)-MS-Ⅱ column(4.6 mm×250 mm, 5 μm) by gradient elution with a mobile phase of methanol-0.2% phosphoric acid aqueous solution at a flow rate of 1.0 mL·min~(-1), detection wavelength of 238 nm, column temperature of 30 ℃, and an injection volume of 10 μL. The results of fingerprint similarity evaluation for 20 batches of salt-fired Eucommiae Cortex indicated that, except for batch S3 with a similarity of 0.893, the similarity of the other 19 batches was of ≥ 0.919, suggesting good similarity. Fourteen common peaks were calibrated and seven common peaks were identified including geniposidic acid. The mass fractions of geniposidic acid, chlorogenic acid, geniposide, genipin, pinoresinol diglucoside, liriodendrin, and pinoresinol-4-O-β-D-glucopyranoside were 0.062 0%-0.426 9%, 0.024 9%-0.116 5%, 0.009 5%-0.052 9%, 0.005 5%-0.034 8%, 0.115 9%-0.317 8%, 0.016 4%-0.108 8%, and 0.026 4%-0.039 8%, respectively. Using CA, PCA, and OPLS-DA, the 20 batches of salt-fired Eucommiae Cortex were classified into three categories. Additionally, through the analysis of variable importance in projection(VIP) under OPLS-DA, two differential quality markers, geniposidic acid and chlorogenic acid, were identified. The established HPLC fingerprint and multi-component content determination method is stable and reliable, providing a reference for quality control of salt-fired Eucommiae Cortex.
本研究建立了盐杜仲的高效液相色谱指纹图谱及多成分含量测定方法,并采用指纹图谱相似度评价、聚类分析(CA)、主成分分析(PCA)和正交偏最小二乘法判别分析(OPLS - DA)对不同来源的盐杜仲质量进行评价。采用Cosmosil 5C₁₈ - MS -Ⅱ色谱柱(4.6 mm×250 mm,5 μm)进行高效液相色谱分析,以甲醇 - 0.2%磷酸水溶液为流动相,梯度洗脱,流速为1.0 mL·min⁻¹,检测波长为238 nm,柱温为30℃,进样量为10 μL。20批盐杜仲指纹图谱相似度评价结果表明,除S3批相似度为0.893外,其余19批相似度均≥0.919,相似度良好。标定了14个共有峰,鉴定了7个共有峰,包括京尼平苷酸。京尼平苷酸、绿原酸、京尼平苷、京尼平、松脂醇二葡萄糖苷、杜仲脂素和松脂醇 - 4 - O -β - D - 葡萄糖苷的质量分数分别为0.062 0% - 0.426 9%、0.024 9% - 0.116 5%、0.009 5% - 0.052 9%、0.005 5% - 0.034 8%、0.115 9% - 0.317 8%、0.016 4% - 0.108 8%和0.026 4% - 0.039 8%。采用CA、PCA和OPLS - DA将20批盐杜仲分为三类。此外,通过OPLS - DA下的变量投影重要性(VIP)分析,鉴定了两个差异质量标志物京尼平苷酸和绿原酸。所建立的高效液相色谱指纹图谱及多成分含量测定方法稳定可靠,为盐杜仲的质量控制提供了参考。