HPLC detection scheme of aflatoxin in hottest feed

HPLC detection scheme of aflatoxin in feed

aflatoxin is a kind of metabolite produced by Aspergillus flavus and parasitic Aspergillus, which has strong toxicity and carcinogenicity, can cause carcinogenesis in many animals, and mainly induce liver cancer. Aflatoxin widely exists in grain, oil and food, among which peanut and corn are the most seriously polluted, and the feed pollution in tropical and subtropical areas is generally heavier. Because of its high toxicity, strong carcinogenicity and universality, it is particularly important to establish a convenient, fast and stable detection method

at present, the national standard methods for aflatoxin detection include thin-layer chromatography, enzyme-linked immunosorbent assay, immunoaffinity column purification high-performance liquid chromatography. Thin layer chromatography is cumbersome, polluting, quantitative and time-consuming; The enzyme-linked immunosorbent assay is simple and sensitive, but its specificity is poor; Immunoaffinity column high performance liquid chromatography is a post column derivatization method, which has a large deviation for samples with high aflatoxin content, and post column derivatization is not popular, which brings inconvenience to use

Saizhi technology application and detection department adopts self-made purification column for purification, pre column derivatization method for aflatoxin derivatization, and uses a new high-performance lc-10tvp high-performance liquid chromatograph and waters 2475 multi-channel fluorescence detector. Through practical detection, it can provide an HPLC detection scheme for the simultaneous determination of Solvay polymer materials in feed to meet the growth needs of the Chinese market. Aflatoxin B1, B2, G1, G2, M1, and the results are accurate and reliable, The detection limit is good. It is a good method for detecting aflatoxin, which is only for the reference of users

the following is a detailed detection method for the determination of aflatoxin in feed

1 instruments and reagents

1.1 instruments

lc-10tvp high performance liquid chromatography

vertex column 150mm 4.6mm 5 m

waters 2475 multi-channel fluorescent detector

self made purification column

1.2 reagents

methanol, chromatographic purity

acetonitrile, chromatographically pure

trifluoroacetic acid

n-hexane

silica gel

neutral alumina

polyamide powder

but all have their plans and objectives. Aflatoxin standard sample: purity 99%

2 preparation of sample solution

2.1 extraction and purification of sample

grind the feed sample with a pulverizer, weigh the ground sample and place it in a flask or stirring bottle, add 84% acetonitrile solution by volume, and vibrate it on a rotating oscillator. Filter it into the sample bottle with a funnel and qualitative filter paper, purify the filtrate through a self-made purification column, discard the previous one, and then transfer the purified solution to a clean brown bottle, and dry it with nitrogen under the condition of water bath at 60 ℃

2.3 pre column derivatization conditions

add n-hexane and trifluoroacetic acid to the residue dried with nitrogen, and screw the cover tightly immediately. Vortex, derivatize in an oven at 40 ℃, dry the mixture with nitrogen at room temperature, dissolve the residue with acetonitrile solution with a volume fraction of 15% and vortex, centrifuge at a speed of 3000 r/min, and inject the purified sample solution into liquid chromatography for analysis

3 chromatographic conditions

chromatographic column: vertex chromatographic column 150mm 4.6mm 5 m

mobile phase: acetonitrile solution

detector: fluorescent detector, wavelength: ex= 360 nm, EM = 440 nm

flow rate: 1.0ml/min

temperature: 25 ℃

injection volume: 20 L

4 liquid chromatogram

using the above liquid chromatographic conditions, five aflatoxins B1, B2, G1, G2, and Jiangsu Changzhou M1, known as the "Oriental carbon Valley" in the industry, can obtain a good separation effect in 10 minutes

add five kinds of aflatoxins to the feed sample containing aflatoxin B1, purify it with a self-made purification column, and then inject the sample for analysis

for more detection schemes, please contact Saizhi technology directly

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