Abstract
This paper describes an immunoaffinity purification technique for 6-keto-prostaglandin F 1α (6KPGF 1α) prior to quantitative analysis by high-resolution gas chromatography—negative-ion chemical ionization mass spectrometry (HRGC—NICIMS). Polyclonal antibodies to 6KPGF 1α 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 6KPGF 1α 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 6KPGF 1α was sufficiently purified using these columns that HRGC—NICIMS analysis of the methoxime—penta-fluorobenzyl—tris-trimethylsilyl derivative was interference-free.
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