通过反相离子对液相色谱法结合柱后放射免疫测定法对神经肽进行表征。应用于促甲状腺激素释放激素、P物质和加压素。

Characterization of neuropeptides by reversed-phase, ion-pair liquid chromatography with post-column detection by radioimmunoassay. Application to thyrotropin-releasing hormone, substance P, and vasopressin.

作者信息

Spindel E, Pettibone D, Fisher L, Fernstrom J, Wurtman R

出版信息

J Chromatogr. 1981 Mar 13;222(3):381-7. doi: 10.1016/s0378-4347(00)84138-9.

Abstract

Neuropeptide contents of rat brain samples were determined by radioimmunoassay (RIA) after fractionation of tissue extracts by high-performance liquid chromatography (HPLC). Solvent systems were composed of acetic acid, acetonitrile and short-chain (5--8 carbons) alkylsulfonic acids. Separate solvent systems were developed for thyrotropin-releasing hormone, substance P. arginine vasopressin and biologic analogs, and the enkephalins. All separation systems tested gave 80--90% recovery of picogram quantities of peptides. When lyophilized, the HPLC solvents did not interfere significantly with the RIAs, allowing quantitation of tissue concentrations of isolated neuropeptides using the lyophilized eluent from the HPLC. The combination of liquid chromatography with RIA should allow for very accurate identification and quantification of peptides in biologic samples containing large numbers of potentially cross-reacting species of molecules.

摘要

通过高效液相色谱法（HPLC）对大鼠脑样本组织提取物进行分级分离后，采用放射免疫分析法（RIA）测定神经肽含量。溶剂系统由乙酸、乙腈和短链（5 - 8个碳）烷基磺酸组成。针对促甲状腺激素释放激素、P物质、精氨酸加压素及其生物类似物以及脑啡肽开发了单独的溶剂系统。所有测试的分离系统对皮克级肽的回收率均为80 - 90%。冻干后，HPLC溶剂对放射免疫分析无显著干扰，从而可以使用HPLC冻干洗脱液对分离出的神经肽的组织浓度进行定量。液相色谱法与放射免疫分析法相结合，应能非常准确地鉴定和定量生物样本中含有大量可能发生交叉反应的分子种类的肽。