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  • P-ISSN 1225-0163
  • E-ISSN 2288-8985

Assay of <TEX>$\beta$</TEX>-Galactosidase Using High Performance Liquid Chromatography

Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
1992, v.5 no.4, pp.465-469
Shin, Myung Gon
Chang, Pahn Shick
Min, Bong Kee
Kim, Sun Chang
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Abstract

An analytical procedure is presented for the quantitative determination of lactose, glucose, and galactose in the hydrolyzate of lactose by <TEX>${\beta}$</TEX>-galactosidase with high-performance liquid chromatography. An Aminex HPX-87C column at <TEX>$85^{\circ}C$</TEX> and refractive index detector were used to resolve lactose, glucose, and galactose in only 12 minutes with distilled and deionized water as a mobile phase. The validity of high-performance liquid chromatography as a method for the assay of <TEX>${\beta}$</TEX>-galactosidase was supported by recovery experiments and comparision of results with those by ONPG method, a spectrophotometric assay. The procedure was appropriate for determination of sugars in the enzyme reaction mixture and could by applied to analysis of <TEX>${\beta}$</TEX>-galactosidase activity.

keywords
<TEX>${\beta}$</TEX>-Galactosidase, HPLC, ONPG, quantitative determination


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