Abstract:Objective To develop an approach for simultaneous detection of multi-mycotoxins in fresh fruits, so as to provide technical supports for mycotoxins surveillance in fresh fruits. Methods Fresh fruits were collected from markets and homogenized. Then, 2 g of fresh fruits were added with 10 mL of 0.1% formic acid ( 99∶1, v/v ) in acetonitrile and wortexed for 10 min. Following extraction with 1 g of sodium chloride and 4 g of anhydrous sodium sulfate, samples were centrifuged and 5 mL of the supernatant was cleaned up with 25 mg C18. Following centrifugation, the supernatant was dried under nitrogen. The residue was dissolved in 300 μL of methanol-acetonitrile mixture solution ( 1∶1, v/v ), and mixed evenly in 700 μL of the distilled water. Samples were then eluted in gradient series of 0.1% formic acid and 5 mmol ammonium formate and methanol-acetonitrile mixture solution ( 1∶1, v/v ). The 15 mycotoxins were determined using liquid chromatography-tandem mass spectrometry ( LC-MS/MS ) with electrospray ion source (ESI+/ESI-) under multiple reaction monitoring. In addition, a matrix-matched standard curve was employed for quantitative analysis. Results There was a good linear relationship for 15 mycotoxins at concentrations of 0.25 to 10 ng/mL ( R2>0.992 ), the LC-MS/MS method showed the detection limits of 0.1-1.0 μg/kg, the spiked recovery rates of 71.68%-117.50%, and the relative standard deviations ( RSDs ) of 0.01%-13.60%. The detection rate of mycotoxins was 27.09% in 203 fresh fruits sold in markets. Conclusions The optimized LC-MS/MS method can be used for simultaneous determination of multi-mycotoxins in fresh fruits.
[1] ESKOLA M,KOS G,ELLIOTT C T,et al.Worldwide contamination of food-crops with mycotoxins:validity of the widely cited‘FAO estimate’of 25%[J].Crit Rev Food Sci Nutr,2022,60(16):2773-2789. [2] WANG Y J,NIE J Y,YAN Z,et al.Multi-mycotoxin exposure and risk assessments for Chinese consumption of nuts and dried fruits[J].J Integr Agric,2018,17(7):1676-1690. [3] SENGLING CEBIN COPPA C F,MOUSAVI KHANEGHAH A,ALVITO P,et al.The occurrence of mycotoxins in breast milk,fruit products and cereal-based infant formula:a review[J].Trends Food Sci Technol,2019,92:81-93. [4] WANG Y J,NIE J Y,YAN Z,et al.Occurrence and co-occurrence of mycotoxins in nuts and dried fruits from China[J].Food Control,2018,88:181-189. [5] ZHOU J,ZHANG D D,CHEN X H,et al.Investigation on the occurrence and contamination of multi-mycotoxin in chestnut and jujube(red date)[J/OL].J Chromatogr A,2021,1659(2021-08-23)[2022-03-22].https://doi.org/10.1016/j.chroma.2021.462486. [6] AROUD H I,MAY B,DIETRICH H,et al.Influence of processing steps on the fate of ochratoxin A,patulin,and alternariol during production of cloudy and clear apple juices[J].Mycotoxin Res,2021,37(4):341-354. [7] SADHASIVAM S,BARDA O,ZAKIN V,et al.Rapid detection and quantification of patulin and citrinin contamination in fruits[J/OL].Molecules,2021,26(15)(2021-07-27)[2022-03-22].https://doi.org/10.3390/molecules26154545. [8] ZHOU J,XU J,CAI Z,et al.Simultaneous determination of five alternaria toxins in cereals using QuEChERS-based methodology[J].J Chromatogr B Analyt Technol Biomed Life Sci,2017,1068/1069:15-23. [9] FAN Y Y,LIU F J,HE W Z,et al.Screening of multi-mycotoxins in fruits by ultra-performance liquid chromatography coupled to ion mobility quadrupole time-of-flight mass spectrometry[J/OL].Food Chem,2022,368(2021-08-13)[2022-03-22].https://doi.org/10.1016/j.foodchem.2021.130858. [10] LAGO L O,NIEVIEROWSKI T H,MALLMANN L P,et al.QuEChERS-LC-QTOFMS for the simultaneous determination of legislated and emerging mycotoxins in malted barley and beer using matrix-matched calibration as a solution to the commercial unavailability of internal standards for some mycotoxins[J/OL].Food Chem,2021,345(2020-12-08)[2022-03-22].https://doi.org/10.1016/j.foodchem.2020.128744. [11] 中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会.合格评定化学分析方法确认和验证指南:GB/T 27417—2017[S].北京:中国标准出版社,2017. General Administration of Quality Supervision, Inspection and Quarantine,Standardization Administration of the People's Republic of China.Comformity assessment-Guidance on validation and verification of chemical analytical methods:GB/T 27417-2017[S].Beijing:Standards Press of China,2017. [12] European Commision.Commission Regulation(EU)No 519/2014 of 16 May 2014 amending Regulation(EC)No 401/2006 as regards methods of sampling of large lots,spices and food supplements,performance criteria for T-2,HT-2 toxin and citrinin and screening methods of analysis[J].Off J Eur Union,2014,147:29-43. [13] JUAN C,MAÑES J,FONT G,et al.Determination of mycotoxins in fruit berry by-products using QuEChERS extraction method[J].LWT,2017,86:344-351. [14] GUO W B,YANG J H,NIU X K,et al.A reliable and accurate UHPLC-MS/MS method for screening of Aspergillus,Penicillium and Alternaria mycotoxins in orange,grape and apple juices[J].Anal Methods,2021,13(2):192-201. [15] ESCRIVÁ L,OUESLATI S,FONT G,et al.Alternaria mycotoxins in food and feed:an overview[J/OL].J Food Qual,2017[2022-03-22].https://doi.org/10.1155/2017/1569748.