蕲蛇毒质量标准及其稳定性研究

Quality Standard and Stability of Agkistrodon Acutus Venom

关键词:  蕲蛇毒  质量标准  稳定性

Key words:  Agkistrodon acutus venom  quality standard  stability

摘要:

目的 探讨蕲蛇毒的质量标准及其稳定性。方法 收集17批不同来源的蕲蛇毒,采用双向免疫扩散法、紫外吸收光谱法鉴别蕲蛇毒,对蕲蛇毒效价、干燥失重指标进行检测,采用双缩脲法对蕲蛇毒中的蛋白质进行含量测定,采用SDS-PAGE法对蕲蛇毒中的蛋白质及多肽分子量的分布情况进行考察,采用加速试验和长期试验考察其稳定性。结果 双向免疫扩散实验中,17批尖吻蝮蛇毒皆可与抗五步蛇毒血清产生沉淀线,说明该方法能够准确鉴别出蕲蛇毒。此外,紫外吸收光谱法可以辅助鉴别尖吻蝮蛇毒,本品在278～280nm的波长处吸收度应在1.30~1.70之间；蕲蛇毒干燥失重不得超过10.0%；以每1mg蛋白含酶活力计,其效价应在40单位~100单位之间；蕲蛇毒条带在10~130KDa范围内均有分布；以人血清白蛋白作对照,蕲蛇毒中蛋白质含量不得少于90.0%；3批蕲蛇毒经6个月加速试验和12个月长期试验考察,各项指标均符合质量标准要求。结论 本研究建立的方法专属性好,准确可靠,可用于蕲蛇毒的质量控制,为规范蕲蛇毒质量控制提供科学依据。

Abstract:

Objective To investigate the quality standard and stability of Agkistrodon acutus venom. Methods Seventeen batches of Agkistrodon acutus venom from different sources were collected, and its were identified by two-way immunodiffusion and ultraviolet absorption spectrometry; potency and loss on drying were examined; the protein content of its were determined by bis-urea; the distribution of molecular weights of the proteins and peptides in AAV were investigated by SDS-PAGE; and the stability of its were examined by accelerated and long term tests. RESULTS In the two-way immunodiffusion experiment, all 17 batches of AAV could produce precipitation lines with Agkistrodon acutus antivenin, indicating that this method can accurately identify AAV. In addition, ultraviolet absorption spectroscopy can assist in the identification of its, and the absorbance of this product at the wavelength of 278-280 nm should be between 1.30 and 1.70; loss of weight on drying of its shall not exceed 10.0%; the enzyme activity per 1 mg of protein, its potency shall be between 40 and 100 units; the bands of its are distributed within the range of 10~130KDa; the protein content of its shall not be less than 90.0% when human serum albumin is used as the control ; three batches of AAV were examined in 6-month accelerated test and 12-month long-term test, and all the indexes met the requirements of quality standard. Conclusion The method established in this study is exclusive, accurate and reliable, and can be used for the quality control of AAV, which provides a scientific basis for standardizing the quality control of its.