Simple and sensitive method for the quantification of total bilirubin in human serum using 3-methyl-2-benzothiazolinone hydrazone hydrochloride as a chromogenic probe.
We here describe a new spectrophotometric method for measuring total bilirubin in serum. The method is based on the cleavage of bilirubin giving formaldehyde which further reacts with diazotized 3-methyl-2-benzothiazolinone hydrazone hydrochloride giving blue colored solution with maximum absorbance at 630 nm. Sensitivity of the developed method was compared with Jendrassik-Grof assay procedure and its applicability has been tested with human serum samples. Good correlation was attained between both methods giving slope of 0.994, intercept 0.015, and R(2)=0.997. Beers law obeyed in the range of 0.068-17.2 μM with good linearity, absorbance y=0.044 C(bil)+0.003. Relative standard deviation was 0.006872, within day precision ranged 0.3-1.2% and day-to-day precision ranged 1-6%. Recovery of the method varied from 97 to 102%. The proposed method has higher sensitivity with less interference. The obtained product was extracted and was spectrally characterized for structural confirmation with FT-IR, ¹H NMR.
Nagaraja
Padmarajaiah
P
Department of Studies in Chemistry, University of Mysore, Manasagangotri, Mysore 570006, India. profpn58@yahoo.com
Avinash
Krishnegowda
K
Shivakumar
Anantharaman
A
Dinesh
Rangappa
R
Shrestha
Ashwinee Kumar
AK
D052160:Benzothiazoles; D001663:Bilirubin / Q000097:blood*; D002863:Chromogenic Compounds; D006801:Humans; D006835:Hydrazones; D008722:Methods; D012680:Sensitivity and Specificity; D013053:Spectrophotometry / Q000379:methods
D052160:Benzothiazoles; D002863:Chromogenic Compounds; D006835:Hydrazones; C002536:3-methyl-2-benzothiazolone hydrazone; D001663:Bilirubin