Analysis of inositol by high-performance liquid chromatography.
A high-performance liquid chromatographic (HPLC) method has been developed for identification and quantification of inositol isomers and monosaccharides in inositol-containing glycans. The method, which can determine 10 pmol of inositol, utilizes an Aminex HPX-87C column packed with an 8% crosslinked cation-exchange resin in the calcium form eluted with deionized water at 50 degrees C. NaOH solution is added to the column effluent through a postcolumn tee to increase the pH (pH greater than 11.6) before entering a pulsed amperometric detector which is highly sensitive for polyhydroxylated compounds. Samples in which inositol is linked to sugar through a glycosidic bond are hydrolyzed with 5.5 N trifluoroacetic acid, 100 degrees C, 4 h, and then reduced with NaBH4. Samples in which inositol is linked via a phosphate ester are hydrolyzed with 6 N HCl, 110 degrees C, 24 h. This method has been applied to the analysis of inositol in the hamster prion proteins (PrP) PrP27-30, and PrPSc.
Wang
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WT
BioCarb Inc., Gaithersburg, Maryland 20879.
Safar
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J
Zopf
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D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006224:Cricetinae; D006863:Hydrogen-Ion Concentration; D007294:Inositol / Q000032:analysis*; D007700:Kinetics; D015394:Molecular Structure; D009005:Monosaccharides / Q000032:analysis; D011134:Polysaccharides / Q000032:analysis; D013237:Stereoisomerism; D013696:Temperature
D009005:Monosaccharides; D011134:Polysaccharides; D007294:Inositol