Quantitative analysis of 6-keto-prostaglandin F1 alpha using immunoaffinity purification and gas chromatography-mass spectrometry.
This paper describes an immunoaffinity purification technique for 6-keto-prostaglandin F1 alpha (6KPGF1 alpha) prior to quantitative analysis by high-resolution gas chromatography-negative-ion chemical ionization mass spectrometry (HRGC-NICIMS). Polyclonal antibodies to 6KPGF1 alpha were partially purified using Staphylococcus aureus Protein A immobilized on Sepharose CL-4B. This partially purified fraction was covalently bound to silica gel using N-hydroxysuccinimidyl-functionalized silica. Columns constructed using this gel quantitatively bound 6KPGF1 alpha which could be eluted quantitatively with acetonitrile-water (19:1). Binding capacity was reconstituted by washing with 0.01 M phosphate buffer (pH 7.4). Human urinary and canine plasma 6KPGF1 alpha was sufficiently purified using these columns that HRGC-NICIMS analysis of the methoxime-pentafluorobenzyl-tris-trimethylsilyl derivative was interference-free.
D015121:6-Ketoprostaglandin F1 alpha / Q000032:analysis*; D002846:Chromatography, Affinity; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D008297:Male; D012822:Silicon Dioxide; D005719:gamma-Globulins / Q000302:isolation & purification
D005719:gamma-Globulins; D015121:6-Ketoprostaglandin F1 alpha; D012822:Silicon Dioxide