Comparison of Paired-Ion Liquid Chromatographic Method with AOAC Fluorometric and Microbiological Methods for Riboflavin Determination in Selected Foods

Abstract

A paired-ion liquid chromatographic (LC) technique coupled with fluorometric detection to determine riboflavin in various food matrices is described. Chromatograms of many foods showed 2 peaks of interest due to presence of riboflavin and flavin mononucleotide (FMN). Relatively high levels of FMN were found in raw beef, corned beef, chicken liver, and canned mushrooms. When riboflavin and FMN contents were summed, LC values were comparable to those obtained by the AOAC standard procedures. The LC technique was sensitive, rapid, and simple, yielding a mean standard deviation of 3.1% which was comparable to the AOAC fluorometric method (3.0%) and better than the AOAC microbiological assay (9.6%). Mean spike recoveries were 91.8% for LC compared to 90.5% and 89.6% for the AOAC fluorometric and microbiological methods, respectively.

Detection and Assay of Vitamin B-2 (Riboflavin) in Alkaline Borate Buffer with UV/Visible Spectrophotometry

“…Utilizing high-performance thin layer chromatography and various mobile phase solvents, such as 1-butanol-chloroform-acetic acid-ammonia-water, 7 : 4 : 5 : 1 : 1, or benzene-methanol-acetone-acetic acid, 70 : 20 : 5 : 5, or chloroform-ethanol-acetone-ammonia, 2 : 2 : 2 : 1, achieved some useful level of separation for riboflavin identification [ 10 ]. Other various liquid chromatography approaches were able to extract vitamins B-1 and B-2 from foods for determination [ 11 , 12 ] and use reverse-phase chromatography with 254 nm detection [ 13 ], biosensor technology [ 14 ], ion-pair chromatography [ 15 , 16 ], determination in dairy products [ 17 ], foodstuffs [ 18 ], detection of casein [ 19 ], total riboflavin in foods [ 20 ], riboflavin in infant formula [ 21 , 22 ], and simultaneous thiamine, riboflavin, and niacin in food [ 23 ]. Methods of a riboflavin assay that utilize UV/visible spectrometry with solvation into aqueous media for speed, ease of application, low cost, high sensitivity, accuracy, and extensive application are very few.…”

Determination of Total Riboflavin in Cooked Sausages

“…For the HPLC conditions, the preferred mode has been reversed-phase C 18 (Finglas and Faulks, 1984;Macrae et al, 1984;Ashoor et al, 1985;Brubacher et al, 1985;Suhara et al, 1985;Johnsson and Branzell, 1987;Ollilainen et al, 1990;Barna, 1991), C 8 (Fellman et al, 1982;Macrae et al, 1984), and C 22 (Lumley and Wiggins, 1981). Some researchers (Toma and Tabekhia, 1979;Reyes et al, 1988;Vidal-Valverde and Reche, 1990;Barna and Dworschak, 1994) used ion-pair reversed phase. We found the latter technique better than the former, in which the rapid elution of riboflavin makes difficult its separation from interferences.…”

“…Even though numerous methods have been reported for the determination of riboflavin content in foods, reversed-phase liquid chromatography has been recommended, employing an ultraviolet detector (Toma and Tabekhia, 1979;Ashoor et al, 1985;Suhara et al, 1985;Vidal-Valverde and Reche, 1990;Barna, 1991;Barna and Dworschak, 1994) or a fluorescence detector (Lumley and Wiggins, 1981;Fellman et al, 1982;Finglas and Faulks, 1984;Macrae et al, 1984;Brubacher et al, 1985;Johnsson and Branzell, 1987;Reyes et al, 1988;Ollilainen et al, 1990), because of the strong ultraviolet light absorption of flavins and their intense fluorescence.…”

Assay of Riboflavin in Sample Wines by Capillary Zone Electrophoresis and Laser-Induced Fluorescence Detection

“…Detailed knowledge of the role of RF in wines can be successfully gained only if a sensitive, reliable, and rapid analytical method is available, which should be applicable to all varieties of wines. Although high-performance liquid chromatography (HPLC) is a popular analytical technique for the determination of RF in foodstuffs and biological samples ( − ), and some reference papers and a book have been published that describe the technology in detail ( − ), the technique suffers from a number of drawbacks. These include the need to use gradient elution to separate the compounds of interest in a reasonable time set, a significant consumption of mobile phase, and unsatisfactory peak resolution, especially when real samples with a relevant number of UV-absorbing constituents, such as red wines, are involved.…”