基于网络药理学和HPLC法建立宠物用复方石淋通片中多指标成分的质量控制方法

Establishment of a Quality Control Method based on Network Pharmacology and HPLC Method for the Quality Control of Multi-indicator Components in Compound Shi-lin-tong Tablets for Pets

关键词:  复方石淋通片  网络药理学  HPLC  多指标成分  含量测定

Key words:  Compound Shi-lin-tong Tablets  network pharmacology  HPLC  multi-index component  content determination

摘要:

建立宠物用复方石淋通片多指标成分含量测定方法。利用网络药理学方法筛选复方石淋通片治疗尿石症、膀胱炎的关键活性成分，结合相关文献及《中国药典》相关要求，确定指标性成分。利用HPLC分析方法，使用CAPCELL PAK C18 MG (S-5) 4.6×250mm色谱柱，柱温30℃，进样量10μL，流速为1.0mL·min-1，检测波长335nm，以乙腈-0.1%甲酸水为流动相，梯度洗脱，建立指标性成分含量测定的方法。通过网络药理学筛选出槲皮素、咖啡酸、绿原酸等十个关键成分。结合《中国药典》及相关文献选出新绿原酸、绿原酸、隐绿原酸、咖啡酸、夏佛塔苷、异夏佛塔苷六种指标性成分。含量测定方法学结果表明：各成分线性关系良好（r≥0.999）；精密度良好，RSD为0.03%~0.21%；重复性良好，RSD为0.70%~2.02%；供试品在24h内稳定性良好，RSD为0.30~2.02%；平均加样回收率（RSD）在98.31%~104.31%（0.36%~2.28%）；三批样品中六种成分的含量分别在0.990~1.434mg·g-1、1.533~2.083mg·g-1、1.311~2.023mg·g-1、0.332~0.353mg·g-1、1.594~1.768mg·g-1、0.787~1.057mg·g-1之间。该含量测定方法有较好的重复性和稳定性，可用于宠物用复方石淋通片的质量分析。

Abstract:

To establish a multi-indicator method for the determination of the content of Compound Shi-lin-tong Tablets for pets. Screening the key active ingredients of Compound Shi-lin-tong Tablet for the treatment of urolithiasis and cystitis by using network pharmacological methods, combining with the relevant literature and the relevant requirements of Chinese Pharmacopoeia, to determine the indicator ingredients. The method was established for the determination of the content of the indicator components by HPLC analysis using a CAPCELL PAK C18 MG (S-5) 4.6×250 mm column at 30℃ with an injection volume of 10μL, a flow rate of 1.0 mL/min, and a detection wavelength of 335nm, with a gradient elution using acetonitrile-0.1% formic acid in water as the mobile phase. Ten key components such as quercetin, caffeic acid, and chlorogenic acid were screened by network pharmacology. Six index components, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, charophylloside and isochlorophylloside, were selected in conjunction with the Chinese Pharmacopoeia and related literature. The methodological results of the content determination showed: The linearity of the components was good (r≥0.999); Good precision, RSD of 0.03%~0.21%; Repeatability was good with RSD of 0.70% to 2.02%; The stability of the test material was good over 24h with RSD of 0.30-2.02%; The average spiked recoveries (RSD) ranged from 98.31% to 104.31% (0.36% to 2.28%); The levels of the six components in the three batches of samples ranged from 0.990 to 1.434mg·g-1, 1.533 to 2.083mg·g-1, 1.311 to 2.023mg·g-1, 0.332 to 0.353mg·g-1, 1.594 to 1.768mg·g-1, and 0.787 to 1.057mg·g-1, respectively. The content determination method has good reproducibility and stability, and can be used for the quality analysis of Compound Shi-lin-tong Tablets for pets.