艾塞那肽缓释微球体外释放度研究及其方法学验证

Research on In Vitro Release of Exenatide Extended-release Microspheres and Method Validation

目的建立艾塞那肽缓释微球的体外释放检测方法,并对其进行方法学研究,验证方法的可靠性及耐用性。方法使用菲罗门Jupiter XB-C18柱(25 cm×4.6 mm,5μm,300?),以0.5%磷酸水/乙腈溶液(80/20)为流动相A,以0.5%磷酸乙腈溶液为流动相B,采用梯度洗脱;检测波长为210 nm;柱温:50℃;流速:1.0 ml/min;进样体积:20μl。采用释放度测定法,通过检测球内剩余药物的量间接测定体外累积释放度。结果计算累积释放度及其相关性表明,释放曲线拟合符合波尔兹曼方程;37和45℃释放度有良好的线性相关性。结论为了缩短样品周期,质量标准中选取45℃释放方法,并通过方法学研究,验证了方法的可靠性、科学性及耐用性。

Objective To develop a method for determination of in vitro release of exenatide extended-release microspheres, and verify the reliability and durability of the method. Methods An HPLC with gradient elution was adopted. Separation was accomplished on C18 column（Phenomenex, Jupiter XB-C18, 25 cm×4.6 mm, 5 μm, 300 ）, using 0.5 % phosphoric acid-water/acetonitrile（80：20） as mobile phase A, and 0.5 % phosphoric acid acetonitrile solution as mobile phase B, with a flow rate of 1.0 ml/min. The detection wavelength was 210 nm, the column temperature was 50 ℃, and the injection volume was 20 μl. The in vitro release of exenatide extendedrelease microspheres was determined indirectly through determination of residual drug in microspheres. Results The cumulative release and its correlation indicated that the release curve fits Boltzmann equation. The release at 37 ℃ and 45 ℃ has good correlation. Conclusions 45 ℃ can be selected as the ideal temperature to detect the in vitro release of exenatide extended-release microspheres. This method is reliable, scientific and durable.