Semi-quantitative method for high molecular weight neuropeptides by high-performance liquid chromatography/thermospray mass spectrometry.
High-performance liquid chromatography/thermospray mass spectrometry (HPLC/MS) was evaluated and optimized for the determination of beta- and gamma-endorphin. Thermospray spectra for the endorphins could be acquired under ion evaporation conditions, when a low vaporizor temperature (94 degrees C) and high source temperature are used. The spectra showed [M + H]+, [M + Hx-1 + Nax]+(x = 1-3) together with double charged ions. Positive ion detection exhibited higher ion currents than negative ion detection. Thermospray HPLC/MS was used to generate a linear calibration curve from 10 to 300 pmol. Thermospray displayed good run-to-run reproducibility (2-7%) and accurately measured spiked quantities of beta- and gamma-endorphin within 15% of the spiked values.
Voyksner
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RD
Analytical and Chemical Sciences, Research Triangle Institute, Research Triangle Park, North Carolina 27709.
Pack
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TW
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D004723:Endorphins / Q000134:cerebrospinal fluid*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D001615:beta-Endorphin / Q000134:cerebrospinal fluid; D018823:gamma-Endorphin
D004723:Endorphins; D001615:beta-Endorphin; D018823:gamma-Endorphin