Optimizing recovery of peptides from an octadecylsilyl (ODS) cartridge.
An analytical procedure has been studied in detail to optimize the recovery of the radiolabeled and unlabeled synthetic peptides substance P and methionine-enkephalin from a reversed phase octadecylsilyl disposable cartridge, which is one of the first steps to preferentially enrich peptides from a biologic matrix. Our previous research on biologic peptides indicated the need for the present study. The column flow-rate was found to be one of the most important experimental parameters; and its effect on the recovery of those two synthetic peptides was determined by the measurement of recovered radioactivity and by high performance liquid chromatography u.v. detection. The accuracy and reproducibility of the flow-rate affected recovery, and it was necessary to regulate the flow-rate with a syringe infusion pump. Recovery was optimum using a slow rate for both sample application and elution. Furthermore, a slow elution was more important than a slow rate of sample application. Such basic recovery studies on individual synthetic peptides are required before undertaking peptide extractions from biologic matrices.
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Charles B. Stout Neuroscience Mass Spectrometry Laboratory, College of Medicine, University of Tennessee, Memphis.
Desiderio
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D002851:Chromatography, High Pressure Liquid; D004209:Disposable Equipment; D004744:Enkephalin, Methionine / Q000138:chemical synthesis / Q000302:isolation & purification*; D007553:Isotope Labeling; D012822:Silicon Dioxide; D013056:Spectrophotometry, Ultraviolet; D013373:Substance P / Q000138:chemical synthesis / Q000302:isolation & purification*; D014316:Tritium
D014316:Tritium; D013373:Substance P; D004744:Enkephalin, Methionine; D012822:Silicon Dioxide