HPLC-ESI／MS法测定大鼠血浆及脑组织中H102肽的浓度

目的建立大鼠血浆和脑组织中H102肽浓度测定的HPLC．ESI／MS法。方法血浆和脑组织样品采用甲醇沉淀蛋白后进样测定。色谱柱：GeminiC18（50mill×2．00lnrn，5／．tm），流动相：乙腈（0．1％甲酸）．水（0．1％甲酸）（14．5：85．5），流速0．4mL min-1，柱温40-12。采用电喷雾正离子模式离子化，选择m／x645．3和m／z843．2分别作为H102肽及内标安普利泰的检测离子。结果血浆及脑组织中H102肽在5—500μg·L-1内线性良好（r=0．9992，0．9993）；日内、日间精密度良好（RSD均〈15％）；提取回收率〉85％。SD舍大鼠鼻腔给予H102肽溶液（2mg·kg-1）后，主要药动学参数：l。。5min，pmax 103．13μg·L-1。大鼠嗅球、大脑、小脑及海马中H102肽的AUC0-60min分别为3149．07＇、266．98、243．72及407．77μg·min·L-1。结论本方法快速、准确、重复性好，适用于大鼠体内H102肽浓度的测定。

Determination of H102 peptide in rat plasma and brain tissues by HPLC-ESI/MS

AIM To establish a HPLC-ESI/MS method for the determination of H102 peptide in rat plasma and brain tissues. METHODS Methanol was used for protein precipitation. The separation was carded out on a Gemini Cls column （50 mmx 2.00 ram, 5 t_tm）with the mobile phase consisted d acetonitrile （0.1% formic acid）-water （0.1% formic acid） （14.5：85.5）, which was pumped at a flow rate of 0.4 mL＂ min-1. The positive ionization mode was cho- sen. The chosen ions were m/z 645.3 for H102 and m/z 843.2 for eptifibatide as internal standard, which were de- tected by electro-spray ionization （ESI）. RESULTS The liner range of H102 in rat plasma and brain tissues were 5 - 500 μg L-1（ r = 0. 999 2,0. 999 3）. The intra-day RSD and inter-day RSD for plasma and brain samples were below 15% and the extraction recovery was more than 85%. The pharmacokinetic parameters after intranasal administration with H102 were as follows： tmax was 5 min,pmax was 103.13 μg L-1. The AUC0-60min of H102 in olfactory bulb, cere- brum, cerebellum and hippoeampus were 3 149.07, 266.98, 243.72 and 407.77 μg min L- 1, respectively. CON- CLUSION A fast, accurate and good reproducible HPLC-ESI/MS method for the determination of H102 in rat plasma and brain tissues is established.