温控型胰岛素液体缓释肛门栓体外释放研究

In vitro release of thermosensitive slow-release rectal liquid suppository of insulin

目的 用高效液相色谱法建立温控型胰岛素液体缓释肛门栓中胰岛素含量测定的方法,以察温控型胰岛素液体缓释肛门栓的体外释放特性.方法 色谱柱：Welch ultimate C18（4.6 mm＆#215;250 mm,5μm）；流动相：0.2 mol/L硫酸盐缓冲液-乙腈（74∶26,乙醇胺调pH到2.3）；检测波长：214 nm;柱温：40℃;流速：1 ml/min,对胰岛素含量进行测定.采用无膜释放模型、牛肠黏膜释放模型、羊肠黏膜释放模型,以胰岛素原料药为模型药物；考察温控型胰岛素液体缓释肛门栓的体外释药机制.结果 温控型胰岛素液体缓释肛门栓在1.0 ～40.0 U/ml范围内线性关系良好（r=0.999 1）,平均回收率99.35％,RSD为0.79％；无膜释放：Q=0.4954t-2.6118（r=0.9977）;牛肠黏膜释放：（1-Q） 1/3 =0.001 6t ＋2.927 4（r=0.975 5）羊肠黏膜释放：lnQ=0.878 3lnt-4.931 1（r=0.9705）.结论 此方法简便,重复性好、专属性强、结果准确、可靠,适用于该制剂中胰岛素含量的测定.可考察温控型胰岛素液体缓释肛门栓机制.

Objective To establish HPLC method for the content determination of insulin in thermosensitive slow-release rec- tal liquid suppository , evaluate release characteristics in vitro. Methods The separation was performed on a Welch ultimate column （4.6 mm× 250 mm ,5 μm）. The mobile phase was composed of 0.2 mol/L sodium sulfate buffer （ pH was adjusted to 2.3 with etha- nolamine）-acetonitrile （74 ： 26） with flow rate lml/min,detected at 214 nm, the insulin levels were determined. Adopting membrane less release model, bovine intestinal mncosa release model, the gut mucosa release model, insulin was used as model drug, drug re- lease meehanism of thermosensitive slow-release rectal liquid suppository of insulin in vitro was evaluate. Results The insulin in situ gel was linear in the range of 1.0-40.0 U/ml（ r = 0. 999 1 ）. The average recovery was 99.35% , RSD was 0.79 ; membrane-less re- lease：Q =0. 495 4t -2. 611 8（r =0. 997 7） ;bovine intestinal mucosa release：（1 - Q） 1/3 = 0. 001 6t ＋ 2. 927 4（r = 0. 975 5） ;gut- mucosa release ：lnQ = 0. 878 31nt - 4.931 1 （ r = 0. 970 5 ）. Conclusion The method was simple, fine repeatability, sensitive, accu- rate and reliable. The method could be applicable to the preparation of the content determination of insulin and evaluate release mecha- nism thermosensitive slow-release rectal liquid suppository of insulin.