校对 ---鹿瓜多肽注射剂质量评价与标准优化

通过探索性研究对鹿瓜多肽注射剂质量现状及存在问题进行评价,并对现行质量标准进行优化。首先,对影响终产品新鲜度的因素进行考察。建立丹磺酰氯柱前衍生-HPLC 法测定鹿瓜多肽注射剂中 8 种生物胺的含量,结果显示个别企业的样品中检出较高浓度的尸胺；采用免疫亲和柱净化-柱后光化学衍生-HPLC -FLD 法测定鹿瓜多肽注射剂及中间提取液中黄曲霉毒素 B₁、B₂、G₁ 和 G₂ 的含量,并采用自动电位滴定法测定甜瓜子原料的酸价和过氧化值,结果表明部分企业生产原料甜瓜子存在霉变、酸败等问题,应在新鲜度方面给予重视。其次,优化了鹿瓜多肽注射剂质量标准中高分子量物质和多肽含量测定的方法。采用 Tricine-SDS-PAGE 电泳法代替凝胶色谱法测定高分子量物质,提高了测定的准确性；采用试剂盒法代替福林酚法测定多肽含量,该法操作简便、专属性更强,适用于大批量样品的快速测定。最后,对鹿瓜多肽注射剂质量标准中缺失的溶血与凝聚、生物学活性测定项目进行研究。建立直接观察法结合酶标仪测定法考察溶血与凝聚反应,结果各企业样品均无溶血与凝聚现象；采用 CCK-8 法考察鹿瓜多肽注射剂对 THP-1 细胞增殖的抑制作用,以此评价其抗炎活性,结果显示鹿瓜多肽注射剂对 THP-1 细胞具有增殖抑制作用,各企业样品的体外抗炎效果具有一定差异。探索性研究结果显示,鹿瓜多肽注射剂原材料、中间产品及终产品的安全性均存在一定隐患,表明本品的整体质量水平一般,且现行质量标准存在缺陷,亟待提高。

Quality evaluation and standard optimization of Cervus and Cucumis polypeptide injection

To evaluate the quality status and existing problems of Cervus and Cucumis polypeptide injection through exploratory research and optimize its current quality standards. First, the factors affecting the freshness of the injection were investigated. Establish dansyl chloride pre-column derivatization-HPLC method to determine the content of 8 biogenic amines in Cervus and Cucumis polypeptide injection. The results showed that relatively higher concentrations of cadaverine have been detected in the products from company B. The contents of aflatoxin B1, B2, G1 and G2 in Cervus and Cucumis polypeptide injection and its intermediate extracts were determined by immunoaffinity column purification, post-column photochemical derivatization and HPLC-FLD methods. The acid value and peroxide value of the melon seeds were determined by automatic potentiometric titration. The results showed that the melon seeds from some companies were with some problems such as rancidity and mildew, which indicated that these companies should pay more attention to the freshness of their products. Secondly, the methods for the determination of high molecular weight substances and polypeptides in the quality standard were optimized. Tricine-SDS-PAGE electrophoresis was used instead of gel chromatography to determine the high molecular weight substances, which improved the accuracy. Compared to the Folin-phenol method, the determination of polypeptide content by the kit is simpler to manipulate and more specific, thus suitable for rapid determination of large batches of samples. Finally, the haemolysis and agglomeration and biological activity of Cervus and Cucumis polypeptide injection were studied. The haemolysis and agglomeration was tested by direct observation combined with microplate reader method, and the results showed that no haemolysis and agglomeration were found in all samples. The anti-inflammatory activity of Cervus and Cucumis polypeptide injection was investigated by its inhibitory effect on THP-1 proliferation in vitro with CCK-8 assay, and the results showed that the inhibitory effect of samples from different companies was different to some extent. In conclusion, certain hidden problems are existing in the raw materials, intermediate extracts and final products of Cervus and Cucumis polypeptide injection, indicating that the overall quality level of this product is not good enough. Furthermore, the current quality standards exhibit defects, required to be improved urgently.