Spectrofluorimetric determination of vitamin B₁ using horseradish peroxidase as catalyst in the presence of hydrogen peroxide

Abstract

In this work a new spectrofluorimetric method for the determination of vitamin B(1), based on the catalytic activity of horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2), has been developed. Non-fluorescent vitamin B(1) was easily converted through catalytic oxidation in alkaline medium into a fluorescent compound, even without exposure to light. The linear range for vitamin B(1) observed was 0.026-16.83 microg/mL (RSD = 1.75%). The correlation coefficient for the calibration curve and limit of detection were found to be 0.9964 and 0.015 microg/mL, respectively. The developed method is practical, simple, sensitive and relatively free from interference by coexisting substances and has been successfully applied for the determination of vitamin B(1) in pharmaceutical preparations.

CHAPTER 15. Spectrofluorimetric Analysis of Vitamin B1 in Pharmaceutical Preparations, Bio-fluid and Food Samples

“…The resulting product from the oxidation reaction of vitamin B 1 and H 2 O 2 exhibits a strong fluorescence emission peak at 382 nm when excited at 368 (shown in Figure 15.2) in the presence of HRP; in the absence of HRP, it shows almost no fluorescence peak (Khan et al 2009). Figure 15.2 implies that the non-fluorescent vitamin B 1 is converted to fluorescent thiochrome in the presence of H 2 O 2 by the catalytic activity of HRP, which enhances the fluorescence intensity significantly.…”

“…Thiamin is oxidized to TC by the formation of thiamin disulfide (TDS) from two thiamin molecules in the presence of oxidants. Several spectrofluorimetric methods have been reported for the determination of thiamin based on the oxidation of thiamin using several oxidants including cyanogen bromide (Fujiwara and Matsui 1953), copper (Pe´rez-Ruiz et al 1992), mercury(II) (Martinez-Lozano et al 1990;Perez-Ruiz et al 2004), cobalt(II) (Jie et al 1993), iron(III) tetrasulfonatophthalocyanine (Chen et al 1999), peroxidate (Amjadi et al 2007), horseradish peroxidase (Khan et al 2009). Fujiwara and Matsui (1953) reported a thiochrome reaction to produce thiochrome from thiamin for the determination of thiamin.…”

“…The non-fluorescent substrate (AH 2 ) can be determined by measuring the fluorescence intensity of the oxidation product (A) of the reaction shown in eqn (1) (Khan et al 2009). The resulting product from the oxidation reaction of vitamin B 1 and H 2 O 2 exhibits a strong fluorescence emission peak at 382 nm when excited at 368 (shown in Figure 15.2) in the presence of HRP; in the absence of HRP, it shows almost no fluorescence peak (Khan et al 2009).…”

Stability-Indicating Photochemical Method for the Assay of Thiamine by Spectrophotometry

“…It is extensively used as a component of single vitamin B complex and multivitamin preparations [2][3][4]. Several analytical methods, including UV and visible spectrophotometric and multivariate analysis [5][6][7], fluorimetric [8][9][10][11][12][13][14][15][16][17][18][19][20][21], high performance liquid chromatography (HPLC) [22][23][24][25][26][27][28][29][30][31][32][33][34][35], UHPLC/MS-MS [36], turbidimetric and nephelometric [37], amperometric [38], and voltammetric methods [39][40][41][42][43][44][45], have been used for the assay of thiamine and its salts in pharmaceutical preparations, food materials and biological fluids. Some of these methods have previously been reviewed [46][47][48]…”

“…iamine undergoes chemical [1,8,9,11,14,16,17,19,[51][52][53][54] and photochemical oxidation [55,56] to form thiochrome (TC) which exhibits a blue fluorescence (440 nm). is has been made as the basis of fluorimetric determination of TH [8,9,11,14,16,17,19,[54][55][56][57][58][59][60].…”