A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms

Mushrooms have been used as part of the average diet and as a nutraceutical for thousands of years due to their immense health benefits. The purpose of this study was to develop a simple, fast, accurate, specific, reproducible, and robust chromatographic method to identify and quantify two water-sol...

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Main Authors: Mohammad F. Hossain, Mamoon Rashid, Rajjit Sidhu, Randy Mullins, Susan L. Mayhew
Format: Article
Language:English
Published: Hindawi Limited 2019-01-01
Series:International Journal of Food Science
Online Access:http://dx.doi.org/10.1155/2019/8716986
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spelling doaj-9e89617497314b219ae7fbd0ea76abdf2020-11-24T21:26:24ZengHindawi LimitedInternational Journal of Food Science2356-70152314-57652019-01-01201910.1155/2019/87169868716986A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in MushroomsMohammad F. Hossain0Mamoon Rashid1Rajjit Sidhu2Randy Mullins3Susan L. Mayhew4Appalachian College of Pharmacy, Oakwood, VA 24631, USAAppalachian College of Pharmacy, Oakwood, VA 24631, USAAppalachian College of Pharmacy, Oakwood, VA 24631, USAAppalachian College of Pharmacy, Oakwood, VA 24631, USAAppalachian College of Pharmacy, Oakwood, VA 24631, USAMushrooms have been used as part of the average diet and as a nutraceutical for thousands of years due to their immense health benefits. The purpose of this study was to develop a simple, fast, accurate, specific, reproducible, and robust chromatographic method to identify and quantify two water-soluble vitamins: thiamine (B1) and riboflavin (B2) in mushrooms. The method employed for qualitative and quantitative analysis of these vitamins was Reversed Phase-High Performance Liquid Chromatography (RP-HPLC) equipped with Ultraviolet–Visible (UV-Vis) Detector. The extraction process involved acid hydrolysis followed by enzymatic dephosphorylation with takadiastase enzyme. Chromatographic separation was achieved with a Shimadzu prominence HPLC system using isocratic elution mode on a Waters Xterra® MS C-18 column (4.6mm × 150mm, 5 μm) integrated with a XBridge® BEH C-18 Guard column (2.1mm × 5 mm, 5 μm). The mobile phase of this study consisted of buffer and methanol in the ratio of 80:20, where the buffer contained sodium-1-hexanesulfonate, glacial acetic acid, methanol, and pH adjusted to 3.0 with diethylamine. Vitamins were detected simultaneously at their lambda max wavelengths B1: 245nm and B2: 268nm using dual-wavelength UV detection technique to get their highest response. The proposed method was found to be specific, linear R>1.0, accurate, precise (% recovery ± SD; B1:104.45±4.5 and B2: 104.88±2.04), sensitive, (limit of detection for B1 and B2 was 0.043 and 0.029 μg/mL, respectively), and robust for mushrooms analysis. No coeluting peaks were observed at the retention time of the vitamins and all the peaks were spectrally homogenous. The standard and sample solutions were found to remain stable at cold temperature for 72 hours. In summary, our data suggest that the proposed method could be used in food industries to monitor the product quality during routine quality control purposes.http://dx.doi.org/10.1155/2019/8716986
collection DOAJ
language English
format Article
sources DOAJ
author Mohammad F. Hossain
Mamoon Rashid
Rajjit Sidhu
Randy Mullins
Susan L. Mayhew
spellingShingle Mohammad F. Hossain
Mamoon Rashid
Rajjit Sidhu
Randy Mullins
Susan L. Mayhew
A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms
International Journal of Food Science
author_facet Mohammad F. Hossain
Mamoon Rashid
Rajjit Sidhu
Randy Mullins
Susan L. Mayhew
author_sort Mohammad F. Hossain
title A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms
title_short A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms
title_full A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms
title_fullStr A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms
title_full_unstemmed A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms
title_sort simplified, specific hplc method of assaying thiamine and riboflavin in mushrooms
publisher Hindawi Limited
series International Journal of Food Science
issn 2356-7015
2314-5765
publishDate 2019-01-01
description Mushrooms have been used as part of the average diet and as a nutraceutical for thousands of years due to their immense health benefits. The purpose of this study was to develop a simple, fast, accurate, specific, reproducible, and robust chromatographic method to identify and quantify two water-soluble vitamins: thiamine (B1) and riboflavin (B2) in mushrooms. The method employed for qualitative and quantitative analysis of these vitamins was Reversed Phase-High Performance Liquid Chromatography (RP-HPLC) equipped with Ultraviolet–Visible (UV-Vis) Detector. The extraction process involved acid hydrolysis followed by enzymatic dephosphorylation with takadiastase enzyme. Chromatographic separation was achieved with a Shimadzu prominence HPLC system using isocratic elution mode on a Waters Xterra® MS C-18 column (4.6mm × 150mm, 5 μm) integrated with a XBridge® BEH C-18 Guard column (2.1mm × 5 mm, 5 μm). The mobile phase of this study consisted of buffer and methanol in the ratio of 80:20, where the buffer contained sodium-1-hexanesulfonate, glacial acetic acid, methanol, and pH adjusted to 3.0 with diethylamine. Vitamins were detected simultaneously at their lambda max wavelengths B1: 245nm and B2: 268nm using dual-wavelength UV detection technique to get their highest response. The proposed method was found to be specific, linear R>1.0, accurate, precise (% recovery ± SD; B1:104.45±4.5 and B2: 104.88±2.04), sensitive, (limit of detection for B1 and B2 was 0.043 and 0.029 μg/mL, respectively), and robust for mushrooms analysis. No coeluting peaks were observed at the retention time of the vitamins and all the peaks were spectrally homogenous. The standard and sample solutions were found to remain stable at cold temperature for 72 hours. In summary, our data suggest that the proposed method could be used in food industries to monitor the product quality during routine quality control purposes.
url http://dx.doi.org/10.1155/2019/8716986
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