title: Simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 and ochratoxin A in breast milk by high-performance liquid chromatography/fluorescence after liquid-liquid extraction with low temperature purification (LLE-LTP).
abstract: The aims of this study were to optimize and validate a methodology for the simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 (AFB1, AFB2, AFG1, AFG2, AFM1) and ochratoxin A (OTA) in breast milk, and to analyze these mycotoxins in samples obtained from human milk banks in the Federal District, Brazil. The optimized analytical method was based on liquid-liquid extraction with low temperature purification (3.25mL of acidified acetonitrile+0.75mL of ethyl acetate), followed by analysis by high-performance liquid chromatography with fluorescence detector (HPLC/FLD) and a photochemical post-column reactor. Limits of quantification (LOQ) ranged from 0.005 to 0.03ng/mL, recoveries from 73 to 99.5%, and relative standard deviations (RSD) from 1.8 to 17.3%. The LLE-LTP extraction method was shown to be simple and cost-effective, since no columns were needed for clean-up. Only 2 of the 224 breast milk samples analyzed were positive for the mycotoxins, both samples containing AFB2 at the LOQ level (0.005ng/mL). The identity of the mycotoxin detected was confirmed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). This result indicates that infants who are fed with breast milk from the milk banks are not at risk from aflatoxin and ochratoxin exposure.
authors.affiliation: Laboratory of Toxicology, Faculty of Health Sciences, University of Brasília, 70910-900 Brasília, DF, Brazil.
authors.forename: Patricia Diniz
authors.initials: PD
authors.lastname: Andrade
authors.forename: Julyane Laine
authors.initials: JL
authors.lastname: Gomes da Silva
authors.forename: Eloisa Dutra
authors.initials: ED
authors.lastname: Caldas
mesh_terms: D000348:Aflatoxins / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003080:Cold Temperature; D005260:Female; D006801:Humans; D007223:Infant; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000379:methods*; D008895:Milk, Human / Q000737:chemistry / Q000382:microbiology*; D009793:Ochratoxins / Q000032:analysis*
chemical_list: D000348:Aflatoxins; D009793:Ochratoxins; C025589:ochratoxin A
keywords: Aflatoxins; Analysis; Breast milk; Liquid–liquid extraction; Ochratoxin A