Sulfate and phosphate analysis in glycoproteins and other biologic compounds using ion chromatography. Application to glycoprotein hormones and sugar esters.
An ion chromatography procedure was devised for the simultaneous determination of phosphate and sulfate in the same sample. In order to eliminate interference from zwitterionic compounds (particularly amino acids and peptides) generated during hydrolysis of the phosphate- or sulfate-containing compounds a pretreatment step with a cation-exchange column was required. The detection of sulfate is approximately twice as sensitive as phosphate on a molar basis. The useful working range for sulfate was 200 pmole to 35 nmole with the ion chromatography employed; the range for phosphate was 400 pmole to 65 nmole. Linearity in this range was very satisfactory. Representative analyses are presented for hydrolyzates of several glycoprotein hormones and sugar sulfates and phosphate esters. Replicate analyses were +/- 3.0% or better. The glycoprotein hormone analyses for sulfate did not indicate whole integers per mole, suggesting mixtures of isohormones as has been found by others using chromatofocusing or isoelectric focussing and immunoassay.
Ward
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Department of Biochemistry and Molecular Biology, University of Texas M.D. Anderson Hospital and Tumor Institute at Houston 77030.
Wen
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Bousfield
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D000818:Animals; D002241:Carbohydrates / Q000032:analysis*; D002852:Chromatography, Ion Exchange; D006023:Glycoproteins / Q000032:analysis*; D006062:Gonadotropins / Q000032:analysis; D006728:Hormones / Q000032:analysis*; D006801:Humans; D006868:Hydrolysis; D007986:Luteinizing Hormone / Q000032:analysis; D010710:Phosphates / Q000032:analysis*; D013431:Sulfates / Q000032:analysis*
D002241:Carbohydrates; D006023:Glycoproteins; D006062:Gonadotropins; D006728:Hormones; D010710:Phosphates; D013431:Sulfates; D007986:Luteinizing Hormone