Measurement of proenkephalin A-derived peptides in biological tissues by high pressure liquid chromatography coupled with amperometric electrochemical detection.

Abstract

We describe an analytic method for the separation and quantitation of a number of proenkephalin A-derived peptides using high pressure liquid chromatography coupled with amperometric electrochemical detection (HPLC-AECD). Initially, we coupled our HPLC separation system with AECD in series with a UV detector for additional confirmation of peak specificity. AECD provided a 10(6)-fold increase in sensitivity over UV detection for these peptides. In addition to Met-enkephalin (ME), ME-Arg, ME-Arg-Phe, ME-Arg-Gly-Leu, Leu-enkephalin (LE) and LE-Arg (Dyn 1-6), we separated and detected the sulfoxides of ME and its extended peptides. Subsequently, we used minor modifications of the isocratic mobile phase to separate and detect enkephalin-related peptides with greater sensitivity and shorter chromatographic run times; each of these mobile phases was used to separate and detect two to three peptides. We have applied this HPLC-AECD methodology to quantitate ME, ME-Arg-Phe, ME-Arg-Gly-Leu and LE in pheochromocytoma tumors.

• Authors: S A Mousa, G R Van Loon
• Journal: Life sciences (Life Sci) Vol. 37 Issue 19 Pg. 1795-802 (Nov 11 1985) ISSN: 0024-3205 [Print] Netherlands
• PMID: 4058253 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Enkephalins
• Protein Precursors
• proenkephalin

Topics

• Adrenal Gland Neoplasms (analysis)
• Chromatography, High Pressure Liquid (methods)
• Electrochemistry
• Enkephalins (analysis)
• Humans
• Pheochromocytoma (analysis)
• Protein Precursors (analysis)