摘要
早期研发抗生素品种的纯度与效价之间的定量关系尚不明确,同时控制HPLC纯度和微生物效价是目前各国药典的共同质控策略。由于效价属于特定计量单位,其量值无法直接溯源至国际单位(SI),因此探讨抗生素效价与纯度间的定量关系,实现利用HPLC法同时测定抗生素的纯度与效价成为当前抗生素质量控制的热点。本研究选择多组分抗生素庆大霉素,通过分别制备庆大霉素C1a、C2、C2a和C1单组分样品,利用NMR、HPLC和微生物检定法确定每个C组分的有效成分纯度与理论效价之间的定量关系:每1 mg庆大霉素C1a纯品相当于1 286.98庆大霉素效价单位;每1 mg庆大霉素C2纯品相当于1 095.74庆大霉素效价单位;每1 mg庆大霉素C2a纯品相当于1 079.52庆大霉素效价单位;每1 mg庆大霉素C1纯品相当于739.61庆大霉素效价单位。进而建立了根据HPLC分析得到的庆大霉素C组分的比例与含量确定庆大霉素效价的方法,实现了庆大霉素HPLC纯度分析与效价测定的统一。此外,证明了在蒸发光散射检测器中庆大霉素诸组分与小诺霉素的响应因子一致,即不需要制备庆大霉素诸组分标准品,仅通过小诺霉素标准品就能准确定量诸庆大霉素组分,为HPLC定量庆大霉素诸组分提供了方便。上述方法有望作为常规的质量控制方法,简化目前药典中的繁琐质控策略。
The quality of some earlier developed antibiotics is usually ensured by the combination of HPLC purity and microbiological potency measurement in the pharmacopoeias of various countries because the relationship between their purity and potency is not clearly quantified. Due to potency is assessed using certain units of measurement, it can not be directly traced to the international system of units （SI unit）. This has become a hotspot in the study of the quantitative relationship between purity and potency of antibiotics. It would be quite an achievement to simultaneously determine both purity and potency using HPLC methods during quality control. This study evaluated a multicomponent antibiotic product, gentamycin, as a test sample. First, pure samples of the C components of gentamycin： C1a, C2, C2a and C1 were prepared, separately. Second, quantitative relationship （theoretical potency） between the purity and potency of each C component of gentamycin were determined using IH NMR, HPLC-ELSD and microbiological assay method. One milligram of gentamycin C1a, C2, C2a and C1 was equal to 1 286.98, 1 095.74, 1 079.52 and 739.61 gentamycin units, respectively. Finally, a method for the determination of gentamycin potency was established based on the proportion and content of C components of gentamycin. The unification of purity and potency for gentamycin was achieved using only HPLC-ELSD. It is also demonstrated that C components of gentamycin and micronomicin produce the same responses under ELSD, which means that it is not necessary to prepare separate reference standards for each C component of gentamycin and that quantitative testing can be performed accurately using only one micronomicin reference standard. This study simplified the previous method for the determination of the content of C components of gentamycin using HPLC-ELSD. The developed method is suitable for regular use as a part of quality control and can simplify the rigmarole quality control procedures provided in current pharmacopeias.
出处
《药学学报》
CAS
CSCD
北大核心
2012年第12期1660-1666,共7页
Acta Pharmaceutica Sinica
基金
supported by National Science and Technology Major Project of the Ministry of Science and Technology of China(Grant No.2010ZX09401-403)
