Given severe health-hazardous effects of aflatoxin B1(AFB1) widely occurring in cereal grains and animal feeds,it is highly urgent to develop analytical methods for its rapid screening.In this work,we proposed a simpl...Given severe health-hazardous effects of aflatoxin B1(AFB1) widely occurring in cereal grains and animal feeds,it is highly urgent to develop analytical methods for its rapid screening.In this work,we proposed a simple and high-throughput method for the determination of AFB1 in millet and buckwheat samples using high performance thin layer chromatography(HPTLC) linked to fluorescence densitometry.The first step was to optimize the solid-liquid extraction for the crude clean-up of the samples.The QuEChERS(Quick,Easy,Cheap,Effective,Robust and Safe) extraction strategy was used and different solvent systems for their extraction efficiency of AFB1 from the samples were evaluated.Then,trichloromethane:ethyl acetate(7:3,V/V) was used as the mobile phase to realize the separation of the targeted compound from background noises on silica gel plates.Quantification was readily performed with densitometry in fluorescence mode.In order to fix the optimal excitation wavelength,spectra scanning ranging 250-400 nm was carried out,revealing that 364 nm light gave the highest signal.With the optimized optical system,high sensitivity to AFB1 was achieved,with a limit of detection(LOD) at 3 μg/kg.Apart from that,good linearity(0.999) was obtained within the range of 1-80 ng/band of AFB 1.To assess the analysis accuracy,2 levels of AFB 1(50 and100 μg/kg) were spiked into real grain samples.The obtained results showed that the recovery rates were within the range of 81.6%-114.0%.The proof-of-concept results of this work evidenced that HPTLC is a promising analytical tool for the screening of mycotoxin in difficult samples.展开更多
Aflatoxin B_(1)(AFB_(1))and deoxynivalenol(DON)are among the most frequently detected mycotoxin in cereals,po sing great challenge to human health.This work proposed a co st-efficient and robust planar chromatography ...Aflatoxin B_(1)(AFB_(1))and deoxynivalenol(DON)are among the most frequently detected mycotoxin in cereals,po sing great challenge to human health.This work proposed a co st-efficient and robust planar chromatography method for the rapid screening AFB_(1) in millet and buckwheat,as well as DON in wheat,corn,barley,and oats,achieving detection of the two common fungal toxins in cereals.First,the sample preparation was realized by a simple extraction with 10 mL of acetonitrile,both containing 4 g of anhydrous MgSO_(4),1 g of NaCl,and25 mg of primary secondary amine adsorbent.After high throughput chromatography,which resulted in the separation of the targeted compound from background noise and derivatization with 10%ethanol solution of AlCl_(3)(only for the visualization of DON).The separation results were recorded using a smartphone in a dark chamber equipped with a 366 nm ultraviolet source.To simplify the analysis,the quantification of the graphical outcome was carried out using the open-source software ImageJ,based on quantifying digitalized pixel values.It was demonstrated that the detection sensitivity(AFB_(1):limit of detection(LOD)=14.2μg/kg,DON:LOD=64.3μg/kg)was adequate to meet the strict maximum residue limit stipulated for both targeted compounds,while calibration curves gave good linearity(AFB_(1):correlation coefficient(R^(2))=0.967 within 1-80 ng/band,DON:R^(2)=0.998 within 50-800 ng/band).Then the method accuracy and precision were evaluated by spikingrecovery experiments,showing that the recovery rates of AFB_(1) was within 76.9%-113.6%and those DON within 91.0%-116.5%,with relative standard deviations of less than 4.9%and 5.8%for AFB_(1) and DON,respectively.The conceptual validation of this study demonstrates that the high performance thin layer chromatography(HPTLC)-image quantification method has effectively addressed the limitation of HPTLC imaging results being confined to preliminary screening without the capability of precise quantification,thereby bridging the gap between imaging and quantification.This method exhibits promising analytical tool characteristics and is suitable for screening mycotoxins in cereal samples.展开更多
As a widely used food preservative,methyl paraben was experimentally evidenced with serious hormonelike adverse effects.Herein,a high performance thin-layer chromatography platformed bioluminescent bioautography and i...As a widely used food preservative,methyl paraben was experimentally evidenced with serious hormonelike adverse effects.Herein,a high performance thin-layer chromatography platformed bioluminescent bioautography and image analysis for the selective quantification and confirmation of methyl paraben was proposed and validated in vinegar and coconut juice.First,the detectability of the bioautography to the analyte on different layer materials was estimated,revealing that normal silica gel was the best choice.After that,the liquid of sample extract and working solution were separated to overcome the background noises due to co-extracted matrices.The separation result was then coupled to the optimized bioautography,enabling instant and straightforward screening of the targeted conpound.For accurate quantification,bioluninescent inhibition pattern caused by the analyte was processed by image analysis,giving useful sensitivity(LOD>16 mg/kg),precision(RSD<10.1%)and accuracy(spike-recovery rate 76.9%-112.2%).Finally,the suspected result was confirmed by determining its MS fingerprint,further strengthening the reliability of screening.展开更多
基金supported by National Key Research and Development Program of China (2021YFF0601902)Shanxi Scholarship Council of China (2021-068)+1 种基金Opening Project of Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs China (SWDSJC2021001)Shanxi Agricultural University High-Level Talent Project (2021XG013)。
文摘Given severe health-hazardous effects of aflatoxin B1(AFB1) widely occurring in cereal grains and animal feeds,it is highly urgent to develop analytical methods for its rapid screening.In this work,we proposed a simple and high-throughput method for the determination of AFB1 in millet and buckwheat samples using high performance thin layer chromatography(HPTLC) linked to fluorescence densitometry.The first step was to optimize the solid-liquid extraction for the crude clean-up of the samples.The QuEChERS(Quick,Easy,Cheap,Effective,Robust and Safe) extraction strategy was used and different solvent systems for their extraction efficiency of AFB1 from the samples were evaluated.Then,trichloromethane:ethyl acetate(7:3,V/V) was used as the mobile phase to realize the separation of the targeted compound from background noises on silica gel plates.Quantification was readily performed with densitometry in fluorescence mode.In order to fix the optimal excitation wavelength,spectra scanning ranging 250-400 nm was carried out,revealing that 364 nm light gave the highest signal.With the optimized optical system,high sensitivity to AFB1 was achieved,with a limit of detection(LOD) at 3 μg/kg.Apart from that,good linearity(0.999) was obtained within the range of 1-80 ng/band of AFB 1.To assess the analysis accuracy,2 levels of AFB 1(50 and100 μg/kg) were spiked into real grain samples.The obtained results showed that the recovery rates were within the range of 81.6%-114.0%.The proof-of-concept results of this work evidenced that HPTLC is a promising analytical tool for the screening of mycotoxin in difficult samples.
基金financially supported by the Opening Project of Key Laboratory of Detection for My cotoxins,Ministry of Agriculture and Rural Affairs,China(SWDSJC2021001)。
文摘Aflatoxin B_(1)(AFB_(1))and deoxynivalenol(DON)are among the most frequently detected mycotoxin in cereals,po sing great challenge to human health.This work proposed a co st-efficient and robust planar chromatography method for the rapid screening AFB_(1) in millet and buckwheat,as well as DON in wheat,corn,barley,and oats,achieving detection of the two common fungal toxins in cereals.First,the sample preparation was realized by a simple extraction with 10 mL of acetonitrile,both containing 4 g of anhydrous MgSO_(4),1 g of NaCl,and25 mg of primary secondary amine adsorbent.After high throughput chromatography,which resulted in the separation of the targeted compound from background noise and derivatization with 10%ethanol solution of AlCl_(3)(only for the visualization of DON).The separation results were recorded using a smartphone in a dark chamber equipped with a 366 nm ultraviolet source.To simplify the analysis,the quantification of the graphical outcome was carried out using the open-source software ImageJ,based on quantifying digitalized pixel values.It was demonstrated that the detection sensitivity(AFB_(1):limit of detection(LOD)=14.2μg/kg,DON:LOD=64.3μg/kg)was adequate to meet the strict maximum residue limit stipulated for both targeted compounds,while calibration curves gave good linearity(AFB_(1):correlation coefficient(R^(2))=0.967 within 1-80 ng/band,DON:R^(2)=0.998 within 50-800 ng/band).Then the method accuracy and precision were evaluated by spikingrecovery experiments,showing that the recovery rates of AFB_(1) was within 76.9%-113.6%and those DON within 91.0%-116.5%,with relative standard deviations of less than 4.9%and 5.8%for AFB_(1) and DON,respectively.The conceptual validation of this study demonstrates that the high performance thin layer chromatography(HPTLC)-image quantification method has effectively addressed the limitation of HPTLC imaging results being confined to preliminary screening without the capability of precise quantification,thereby bridging the gap between imaging and quantification.This method exhibits promising analytical tool characteristics and is suitable for screening mycotoxins in cereal samples.
基金financially supported by National Natural Science Foundation of China (21804058)Shanxi Postdoc Reward (SXBYKY2022001)+1 种基金Shanxi Scholarship Council of China (2021068)Shanxi Agricultural University High-Level Talent Project (2021XG013)。
文摘As a widely used food preservative,methyl paraben was experimentally evidenced with serious hormonelike adverse effects.Herein,a high performance thin-layer chromatography platformed bioluminescent bioautography and image analysis for the selective quantification and confirmation of methyl paraben was proposed and validated in vinegar and coconut juice.First,the detectability of the bioautography to the analyte on different layer materials was estimated,revealing that normal silica gel was the best choice.After that,the liquid of sample extract and working solution were separated to overcome the background noises due to co-extracted matrices.The separation result was then coupled to the optimized bioautography,enabling instant and straightforward screening of the targeted conpound.For accurate quantification,bioluninescent inhibition pattern caused by the analyte was processed by image analysis,giving useful sensitivity(LOD>16 mg/kg),precision(RSD<10.1%)and accuracy(spike-recovery rate 76.9%-112.2%).Finally,the suspected result was confirmed by determining its MS fingerprint,further strengthening the reliability of screening.
