基于体外补血活性测定的当归品质评价新方法探索

  目的  建立一种基于补血活性的生物学评价方法, 通过与当归的主要药效相关联, 对其内在品质进行评价。  方法  以氯化血红素为标准品, 用联苯胺染色法测定当归作用于K562细胞后的血红蛋白含量, 通过量反应平行线测定法计算效价, 完成方法学考察并以此比较不同产地、不同炮制方法当归饮片的效价。  结果  当归样品可靠性检验显示, 回归项有显著差异(P＜0.01);方法学考察中重复性和精密度RSD分别为1.13%、4.73%;不同产地当归饮片补血效价为399.61~8 920.32 U·mg-1, 不同炮制方法当归饮片补血效价排序为生当归＞土炒当归＞酒当归, 不同入药部位当归饮片补血效价排序为当归身＞当归头＞当归须。  结论  所建立的当归饮片补血生物效价方法稳定可靠, 方法学考察结果符合《中国药典》2020年版“中药生物活性测定指导原则”相关要求, 可用于不同产地、不同炮制方法、不同入药部位的当归饮片补血疗效的评价, 为进一步完善当归质量评价与控制提供了新思路和新方法。 

Exploration of A New Method for Evaluating the Quality of Angelica Sinensis Based on the Determination of Blood Tonic Activity in vitro

  OBJECTIVE  To develop a biological evaluation method based on the blood tonic activity, combined with the main medicinal effects of Angelica sinensis, to evaluated its internal quality.  METHODS  Hemin was used as standard substance, the relative content of hemoglobin after added Angelica sinensis to K562 cells was determined by the benzidine staining method and the biological titer was calculated by parallel-line assay. After methodological investigation, the biological titers of different production areas and processing of Angelica sinensis were compared.  RESULTS  The reliability test of Angelica sinensis showed that the regression items were significantly different (P < 0.01). The RSDs of repeatability and precision were 1.13% and 4.73%, respectively, in the methodological investigation. The blood-enriching titers of its pieces from different origins were in the range of 399.61-8 920.32 U·mg-1. The rank of blood tonic effects of different processed Angelica sinensis products was raw > soil > wine, and that of different medicinal parts was body > head > tail.  CONCLUSION  The established biological titer method of blood tonic for Angelica sinensis is stable and reliable. The results of methodological investigation are in accord with the relevant requirements of the "Guidelines for the determination of biological activity of traditional Chinese medicines" in the Chinese Pharmacopoeia (2020 edition). It can be used to evaluate the blood tonic titer of Angelica sinensis from different areas, processing methods and medicinal parts, which provides a new idea for further study on the improving of Angelica sinensis quality control.