title: Screening Natural Content of Water-Soluble B Vitamins in Fish: Enzymatic Extraction, HILIC Separation, and Tandem Mass Spectrometric Determination.
abstract: Despite the potential of LC with tandem MS (MS/MS) in improving sensitivity and selectivity, analytical methods are scarce for the determination of protein-bound and phosphorylated forms of B vitamins in food. This prompted us to develop a method for LC-MS/MS determination of naturally occurring nicotinamide, nicotinic acid, thiamine, pyridoxine, riboflavin, pantothenic acid, biotin, folic acid, and cyanocobalamin in fish. Baseline separation of the vitamins was achieved in a hydrophilic interaction LC condition. An ultrasonication-assisted enzymatic extraction protocol for sample preparation was optimized and validated. The time required for extraction was significantly reduced (to 4 h), while maintaining good extraction efficiency. Acetonitrile content (80%, v/v) in the prepared sample was found to be optimum for excellent peak shape and sensitivity. The dynamic linear range of the vitamins ranged from 2.5 to 500 ng/g, and the regression coefficient values were greater than 0.99. LOQ values ranged from 0.4 to 50 ng/g for the different vitamins. The spike recovery values at 50 and 100 ng/g ranged from 87.5 to 97.5%. The intra- and interday precision values were satisfactory. Accuracy of the developed method was determined by analysis of a Certified Reference Material. The method could also be used for unambiguous determination of the natural content of the target vitamins in fish.
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: Niladri Sekhar
authors.initials: NS
authors.lastname: Chatterjee
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: K Ashok
authors.initials: KA
authors.lastname: Kumar
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: K K
authors.initials: KK
authors.lastname: Ajeeshkumar
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: K R Remya
authors.initials: KRR
authors.lastname: Kumari
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: K V
authors.initials: KV
authors.lastname: Vishnu
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: Rangasamy
authors.initials: R
authors.lastname: Anandan
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: Suseela
authors.initials: S
authors.lastname: Mathew
authors.affiliation: Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri PO, Willingdon Island, Cochin-682 029, Kerala, India.
authors.forename: C N
authors.initials: CN
authors.lastname: Ravishankar
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D005399:Fishes; D017747:Seafood / Q000032:analysis*; D053719:Tandem Mass Spectrometry; D014803:Vitamin B Complex / Q000032:analysis*
chemical_list: D014803:Vitamin B Complex