TY - JOUR
T1 - 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
AU - Spindel, Eliot
AU - Pettibone, Douglas
AU - Fisher, Laurel
AU - Fernstrom, John
AU - Wurtman, Richard
N1 - Funding Information:
These studies were supported in part by grants from the National Institutes of Health (HD 11722) and The Center for Brain Sciences and Metabolism Charitable Trust. We thank Dr. S.E. Leeman for providing the antisubstance P serum used in these studies. Dr. Spindel and Dr. Pettibone were recipients of fellowships from the Whittaker Foundation. Dr. Fisher was the recipient of a predoctoral fellowship from NIMH.
PY - 1981/3/13
Y1 - 1981/3/13
N2 - 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.
AB - 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.
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U2 - 10.1016/S0378-4347(00)84138-9
DO - 10.1016/S0378-4347(00)84138-9
M3 - Article
C2 - 6164686
AN - SCOPUS:0019422756
SN - 0378-4347
VL - 222
SP - 381
EP - 387
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
IS - 3
ER -