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Title	Discrete analysis of bile acid in serum and bile with 3⍺-hydroxysteroid dehydrogenase and diaphorase immobilized onto alkylamine glass beads

Creator	Rani, Kirti Garg, P Pundir, C S

Subject	Bile acid 3α-hydroxysteroid dehydrogenase Diaphorase Immobilization Alkylamine glass beads Serum Bile Gallstone

Description	98-104 3α-Hydroxysteroid dehydrogenase (3α-HSD) from Pseudomonas testosteronei and diaphorase (lipoyl dehydrogenase) from Clostridium spp were immobilized individually onto alkylamine glass beads through glutaraldehyde coupling. A cost-effective enzymic colorimetric method for determination of bile acid in the serum and bile was developed employing mixture of the immobilized enzymes. The method was based upon measurement of NADH generated from NAD⁺ during oxidation of bile acid by immobilized 3⍺-HSD with a color reagent consisting of nitrobluetetrazolium (NBT) chloride salt and immobilized diaphorase in 0.065 M sodium phosphate buffer (pH 7.0). The minimum detection limit of the method was 4.8 μmol/L in the serum and 19.5 μmol/L in bile. The per cent recovery of added bile acid in the serum and bile was 89.1 and 95.0, respectively. Within and between batch coefficients of variation (CV) for bile acid determination were

Date	2009-02-25T04:00:06Z 2009-02-25T04:00:06Z 2006-04

Type	Article

Identifier	0301-1208 http://hdl.handle.net/123456789/3263

Language	en_US

Publisher	CSIR

Source	IJBB Vol.43(2) [April 2006]

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