CN103018377A - Enrichment method for detecting aflatoxin B1 by liquid chromatography - Google Patents
Enrichment method for detecting aflatoxin B1 by liquid chromatography Download PDFInfo
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- CN103018377A CN103018377A CN2012105073902A CN201210507390A CN103018377A CN 103018377 A CN103018377 A CN 103018377A CN 2012105073902 A CN2012105073902 A CN 2012105073902A CN 201210507390 A CN201210507390 A CN 201210507390A CN 103018377 A CN103018377 A CN 103018377A
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Abstract
The invention relates to an enrichment method for detecting aflatoxin B1 by liquid chromatography. The enrichment method is characterized by comprising the following steps of: (1) preparing an aflatoxin B1 enrichment material; and (2) adding the aflatoxin B1 enrichment material into a methanol aqueous solution of a sample to be detected, wherein the methanol content in the system is not more than 12.5 percent of the total volume of the system according to volume percentage, and a ratio of the mass of the aflatoxin B1 enrichment material to the total volume of the system is at least 0.625 g/mL, mixing at room temperature, oscillating for 10 to 30 min to enrich the aflatoxin B1, centrifuging, eliminating supernate, diluting with methanol to dilute the aflatoxin B1 enriched in the aflatoxin B1 enrichment material to the methanol, and obtaining a methanol diluted solution containing the aflatoxin B1 for liquid chromatography detection. The enrichment method can be applied to enrichment of liquid chromatography detection of the aflatoxin B1 and is operated easily and quickly; and a result is reliable.
Description
Technical field
The invention belongs to field of biological detection, be specifically related to a kind of AFB
1The liquid chromatographic detection enrichment method.
Background technology
Aflatoxin is the high toxicity secondary metabolite that a class is produced by aspergillus flavus and aspergillus parasiticus secretion.Since " turkey event " beginning of nineteen sixty, people begin to pay close attention to aflatoxin, and the aflatoxin (AFT) of finding now has tens kinds, wherein with AFB
1The most common, toxicity is the strongest.Aflatoxin extensively is present in the agricultural product such as corn, peanut, cottonseed, almond, fig and the food, and its pollution condition can occur in a plurality of links before, during and after the production process in addition, so that it becomes grave danger of agricultural industry and food security.Various standards are set up to total aflatoxin content (AFB in food and the agricultural product in countries in the world for this reason
1, B
2, G
1And G
2Total amount) and AFB
1Maximum allow content all to do mandatory provision.Total aflatoxin content must not surpass 20 μ g/kg, AFB in China GB GB 2761-2011 regulation peanut and the goods thereof
1Content must not surpass 5 μ g/kg.Therefore be present scientific research personnel's research emphasis to the exploitation of Determination Methods of Aflatoxins in food and the agricultural product.
Existing Determination Methods of Aflatoxins mainly contains three kinds of thin layer chromatography, liquid phase chromatography, euzymelinked immunosorbent assay (ELISA).Thin layer chromatography is as classic method, because need operating personnel to be exposed to for a long time in organic solvent and the toxin and be eliminated gradually.Enzyme-linked immunosorbent assay has overcome thin layer chromatography the experimenter has been injured large shortcoming as up-to-date analytical approach, and detects quick and conveniently, and development is rapidly in 20 years in the past.But this method relates to the antibody strong to temperature sensitivity, and common a kind of antibody can only be measured a kind of aflatoxin, in the situation that multiple aflatoxin is to be checked, may there be cross reaction and causes testing result inaccurate, all restricted the promotion and application of the method.So, at present can be by analytical approach or the liquid phase chromatographic analysis method of everybody approval, it is high that it has automaticity, and quantitative and qualitative is accurate, detects the polynary characteristics of kind, is countries in the world official detection method always.But the high efficiency liquid phase chromatographic analysis method for aflatoxin all adopts immune affinity column to come the enrichment toxin at present, and the shortcoming of the antibody that enzyme-linked immunosorbent assay is mentioned still can't be avoided, and therefore studies a kind of new AFB
1Enrichment method to be breaking away from the shortcoming of antibody, and can be at AFB
1Use in the survey, for the AFB in monitoring food and the agricultural product
1Very important meaning and using value.
Summary of the invention
Technical matters to be solved by this invention is the deficiency for above-mentioned prior art existence, and a kind of AFB of being used in the aflatoxin liquid chromatographic detection detects is provided
1The liquid chromatographic detection enrichment method.This enrichment method can be used for AFB
1Enrichment during liquid chromatographic detection is processed, and is simple to operate, fast, and reliable results.
The present invention is that to solve the problems of the technologies described above the technical scheme of employing as follows:
A kind of AFB
1The liquid chromatographic detection enrichment method is characterized in that: it may further comprise the steps:
(1) AFB
1The preparation of enrichment material:
In crystalline flake graphite, add nitration mixture and potassium permanganate that sulfuric acid and phosphoric acid form, carry out preliminary oxidation reaction, then form the graphite oxide that expands by hydrolysis reaction, repeatedly wash successively purifying with hydrogen peroxide, hydrochloric acid and deionized water and obtain graphite oxide afterwards, then with the graphite oxide behind the purifying by the high-intensity ultrasonic mono-layer graphite oxide alkene that is dissociated into, obtain through the centrifugal 2000rpm of choosing not sedimentation in centrifugal 30 minutes and in the centrifugal 30min sedimentation of 5000rpm again;
(2) with the AFB of above-mentioned preparation
1Enrichment material adds in the methanol aqueous solution of testing sample, the content of methyl alcohol the highest 12.5% of the system cumulative volume that do not exceed of percentage meter by volume in the described system, described AFB
1The quality of enrichment material will reach 0.625g/mL with the system cumulative volume than at least, mixed at room temperature, and 10-30min is to carry out AFB in concussion
1Enrichment, centrifugal, remove supernatant, then carry out wash-out with methyl alcohol, with AFB
1The AFB of enrichment in the enrichment material
1Be eluted in the methyl alcohol, obtain containing AFB
1Meoh eluate, liquid chromatographic detection is for subsequent use.
Press such scheme, the system of described step (2) adds water when needed dilutes, so that methanol content the highest 12.5% of the system cumulative volume that do not exceed of percentage meter by volume in the system; The methanol aqueous solution consumption of described testing sample is 4-8mL.
Press such scheme, AFB in the described step (2)
1The quality of enrichment material is preferably 0.625-0.75g/mL with system cumulative volume ratio.
Press such scheme, AFB in described step (2) system
1Content should be in the 0.8-100ng/mL scope, as exceed, can carry out the methanol aqueous solution of testing sample carrying out again enrichment after the corresponding concentrated or dilution process.
Press such scheme, the methanol-eluted fractions step is in the described step (2): with the methyl alcohol AFB that joined enrichment
1AFB
1In the enrichment material, mixing, centrifugal, collect supernatant, be and contain AFB
1Meoh eluate.
Press such scheme, the methanol-eluted fractions step also comprises the repetition elution step in the described step (2), namely adds methyl alcohol in the aforementioned centrifugal precipitation that obtains, and mixing is centrifugal, and supernatant and the aforementioned supernatant that obtains merged, and obtains containing AFB
1Meoh eluate.
Press such scheme, described step (2) also comprises blows the methyl alcohol in the meoh eluate near doing, and adds trifluoroacetic acid and normal hexane and derives, and then adds mobile phase and carries out constant volume, for subsequent use.
Press such scheme, described AFB
1The mass ratio of potassium permanganate and crystalline flake graphite is preferably 6:1 in the preparation of enrichment material; The volume ratio of sulfuric acid and phosphoric acid is preferably 9:1; The massfraction of described graphite in preliminary oxidizing process is preferably 0.70% ~ 0.75%, and the massfraction in the end reaction system after hydrolysis is preferably 0.3% ~ 0.375%; Described preliminary oxidation reaction condition is preferably 50 ℃ of stirring reactions 12 ~ 48 hours.
Press such scheme, in the described step (1) ultrasonic power be 800W or more than, ultrasonic time be 1h or more than.
Preferred 2000rpm not sedimentation in centrifugal 30 minutes and screen in the centrifugal 30min sedimentation of 5000rpm in the above-mentioned steps (1), can avoid centrifugation rate to cross low and may screen the graphene oxide lamella that obtains may be too large or too thick, even may also contain completely graphite of not oxidation, and then cause it to be used for the testing sample AFB
1Adsorption effect not good, can avoid again that centrifugation rate is too high and the graphene oxide lamella that obtains of screening is less, and AFB may cause last desorption the time
1The enrichment material wash-out is incomplete.
This AFB
1The enrichment method AFB
1The liquid chromatographic detection principle of work of enrichment: the inventive method adopts suitable AFB by a large amount of research experiments
1Enrichment material, and suitable methanol content scope in definite system, AFB
1The parameter areas such as the consumption that enrichment material is suitable and enrichment time, and reach based on AFB
1Enrichment material has high-specific surface area and adsorption activity preferably, and directly adsorption of aflatoxin is containing AFB
1The methanol aqueous solution of certain content in enrichment AFB preferably
1Effect, simultaneously because AFB
1Be soluble in the methyl alcohol, utilize the high characteristics of its dissolubility in methyl alcohol with methanol-eluted fractions and with AFB
1From AFB
1Elute in the enrichment material, reach the liquid chromatographic detection AFB
1The purpose of enrichment.
Beneficial effect of the present invention:
(1) AFB provided by the invention
1Liquid chromatographic detection is simple to operate with enrichment method, and fast, reliable results, whole process only need about 30 minutes/sample.
(2) with low cost, and can allow operating personnel that batch sample is processed simultaneously, be easy to popularize.
(3) use therein aspergillus flavus poison toxin B
1The enrichment material stable performance.Compare with antibody or immune affinity column, it does not need special storage condition, does not need other special instrument non-productive operations during enrichment yet.
(4) pollution is few, safe.Organic reagent and other toxic reagents that this enrichment method is related are few, have greatly reduced the pollution to operating personnel and environment.
Embodiment
Embodiment 1 AFB
1Liquid chromatographic detection is determined with enrichment condition in the enrichment method:
(1) AFB
1The preparation of enrichment material
The 3.0g crystalline flake graphite crossed join 400 mL sulfuric acid behind 325 mesh sieves and the phosphoric acid volume ratio is in the mixed solution of 9:1, stirred 10 minutes, then slowly add 18.0g potassium permanganate in potpourri, each addition is too much unsuitable, avoids temperature of reaction to surpass 20 ℃.After potassium permanganate all adds, be heated to 50 ℃ of stirring reactions 12 hours.After reaction finishes, allow potpourri naturally cool to room temperature, be poured into ~ frozen water of 400mL in, continue stirring at room 0.5 hour, be that 30% hydrogen peroxide dropwise adds reaction system and changes glassy yellow into to solution and stop at last with massfraction.Standing over night, outwell supernatant, be the salt pickling 3 times of 5%-10% with the 1L massfraction, then with 1L deionization washing 5 times, to remove metallic ion, sulfate ion and chlorion with the solid freeze drying that obtains, then place deionized water at last, be made into the aqueous solution of 1 mg/mL, use ultrasonic echography 1h, ultrasonic power is 800W, treats that graphene oxide is dissociated into mono-layer graphite oxide alkene, with hydro-extractor 2000 rpm centrifugal 30 minutes, remove sediment, then with centrifugal 30 minutes of supernatant 5000 rpm, remove supernatant, to precipitate again 50 ℃ of dryings, obtain enrichment material.
For ease of using, generally can in enrichment material, add the deionized water stirring at room it is mixed with the aqueous solution that content is 10mg/mL.
(2) AFB
1Liquid chromatographic detection is determined with enrichment condition in the enrichment method:
Under the condition of suitable methanol content (percent by volume is 12.5%), set the consumption (2-6mg) of different aflatoxin B1 enrichment materials in enrichment time (30min) by default suitable length and the system, study AFB
1The amount ranges of enrichment material, specific as follows:
The aflatoxin B1 enrichment material adds with solid form or aqueous solution form all can.Consider following AFB
1Enrichment material adds with solid form, and its impact on the system cumulative volume can be ignored, so add AFB
1AFB before the enrichment material
1The methanol-water standard solution in the content of methyl alcohol be the content of methyl alcohol in the system, add equally AFB
1AFB before the enrichment material
1The methanol-water standard solution in the content of aflatoxin B1 be the content of aflatoxin B1 in the system.When the aflatoxin B1 enrichment material adds fashionablely with the aqueous solution form, the volume of aflatoxin B1 enrichment material solution need be counted in the system cumulative volume.
1. about AFB
1The consumption of enrichment material
With AFB
1Powder standard items (sigma purchase) are dissolved in and are made into the AFB that concentration is 10 ng/mL in the methyl alcohol
1Titer.Get respectively this titer of 1ml, add the 7ml pure water, be mixed to get 5 parts of 8ml and contain the 10ng AFB
1The methanol-water titer.Then contain the 10ng AFB at these 5 parts
1The methanol-water titer in respectively add 2mg, 3mg, 4mg, 5mg and 6mg aflatoxin B1 enrichment material, mixed at room temperature, the concussion 30min, centrifugal 30 minutes of 8000 rpm remove supernatant, add 2ml methyl alcohol, mixing, centrifugal, collect supernatant, and in the aforementioned centrifugal precipitation that obtains, add methyl alcohol, mixing, centrifugal, this supernatant that obtains and aforementioned supernatant are merged, namely contain AFB
1Meoh eluate, nitrogen blows, derivative and upper machine testing AFB
1Content.The recovery is by the AFB that records
1Actual value and known AFB
1The ratio of content value 10ng is tried to achieve, concrete AFB
1The relation of enrichment material consumption and the recovery as shown in Figure 1.As shown in Figure 1: work as AFB
1When the consumption of enrichment material was increased to 5mg, the recovery reached platform area, and namely enrichment material content will reach 5mg at least.
Then the variation of methanol content is tested as follows on the impact of the recovery in the concrete research system:
2. about the content of methyl alcohol in the system
Get the AFB of 1ml 10 ng/mL
1Titer adds the 14ml pure water, is mixed to get 15ml and contains the 10ng AFB
16.25% methanol-water standard solution.Get this titer of 1ml, add the 7ml pure water, be mixed to get 8ml and contain the 10ng AFB
112.5% methanol-water standard solution.Get this titer of 2ml, add the 6ml pure water, be mixed to get 8ml and contain the 10ng AFB
125% methanol-water standard solution.Get this titer of 3ml, add the 5ml pure water, be mixed to get 8ml and contain the 10ng AFB
137.5% methanol-water standard solution.Get this titer of 4ml, add the 4ml pure water, what be mixed to get 8ml contains the 10ng AFB
150% methanol-water standard solution.Contain AFB at above-mentioned 5 parts respectively
1The methanol-water of different proportion treat to add in the enrichment titer 5mg aflatoxin B1 enrichment material, mixed at room temperature, concussion 30min, centrifugal 30 minutes of 8000 rpm remove supernatant, add 2ml methyl alcohol, mixing, centrifugal, collect supernatant, and in the aforementioned centrifugal precipitation that obtains, add methyl alcohol, mixing, centrifugal, this supernatant that obtains and aforementioned supernatant are merged, namely contain AFB
1Meoh eluate, then nitrogen blow, derivative and upper machine testing aflatoxin content.The recovery is by the actual value that records and known AFB
1The ratio of content value 10ng is tried to achieve.AFB
1Methanol aqueous solution in methyl alcohol ratio and the recovery relation as shown in Figure 2.Show in conjunction with Fig. 2: when enrichment material was 5mg, when the methanol content for the treatment of pregnant solution was low to moderate 12.5% gradually by 50%, the recovery reached platform area, then required to treat the enrichment AFB
1The content of the methyl alcohol in the standard solution is the highest must not to surpass 12.5%.
Except this, about the enrichment time research of this enrichment method, specifically test as follows:
3. about enrichment time
Get respectively the AFB of 1ml 10ng/mL
1Titer adds the 7ml pure water, is mixed to get 5 parts of 8ml and contains the 10ng AFB
112.5% methanol-water standard solution.At the above-mentioned AFB that contains
1Methanol-water treat to add 5mg aflatoxin B1 enrichment material in the titer of enrichment, mixed at room temperature is shaken respectively 5min, 10min, 20min, 30min and 40min, centrifugal 30 minutes of 8000 rpm remove supernatant, add 2ml methyl alcohol, mixing, centrifugal, collect supernatant, and in the aforementioned centrifugal precipitation that obtains, add methyl alcohol, mixing, centrifugal, this supernatant that obtains and aforementioned supernatant are merged, namely contain AFB
1Meoh eluate, nitrogen blows, derivative and upper machine testing aflatoxin content.The recovery is by the AFB that records
1Actual value and the known scale value AFB that adds
1The ratio of content value 10ng is tried to achieve, and the relation of concrete enrichment time and the recovery as shown in Figure 3.In conjunction with Fig. 3 as can be known: the recovery is best when enrichment time reaches 10min, reach 30min after the recovery then reach platform area, namely enrichment Best Times section is 10-30min.
After enrichment condition is tentatively determined, about AFB in this enrichment method system applicatory
1Content range has carried out concrete research:
4. with AFB
1Powder standard items (sigma purchase) are dissolved in and are made into the AFB that concentration is respectively 0.8,1,5,10,20,50,100 ng/mL in the methyl alcohol
1Titer.Respectively get the above-mentioned titer of 1ml, add the 7ml pure water, be mixed to get 8ml and contain different amount AFBs
112.5% methanol-water standard solution.Respectively at the above-mentioned 7 parts AFBs that contain different amounts
1Methanol-water treat in the enrichment titer to add 5mg aflatoxin B1 enrichment material, mixed at room temperature, concussion 30min, centrifugal 30 minutes of 8000 rpm remove supernatant, add 2ml methyl alcohol, mixing, centrifugal, collect supernatant, and in the aforementioned centrifugal precipitation that obtains, add methyl alcohol, mixing, centrifugal, this supernatant that obtains and aforementioned supernatant are merged, namely contain AFB
1Meoh eluate, then nitrogen blow, derivative and upper machine testing aflatoxin content.The recovery is by the actual value that records and known AFB
1The ratio of content value is tried to achieve.AFB
1Methanol aqueous solution in methyl alcohol ratio and the recovery relation as shown in Figure 4.Show in conjunction with Fig. 4: when enrichment material was 5mg, when treating that methanol content is lower than 12.5% in the pregnant solution system, the recovery was all between 80-120%.
Embodiment 2 AFBs
1Liquid chromatographic detection uses enrichment method at mark-on peanut sample AFB
1Application in the liquid chromatographic detection:
(1) AFB
1The preparation of enrichment material:
Adopt embodiment 1 described method to prepare enrichment material;
(2) get the blank peanut sample of 5g and add a certain amount of 100 μ L AFBs
1Mark liquid (methanol solutions of 531.25 ng/mL concentration) is placed half an hour, treats that the methyl alcohol volatilization is dried.Then add 15mL 70% methanol-water, the vortex mixing, 50 ℃ ultrasonic 12 minutes, quantitative filter paper filters, get filtrate 4mL and add 2m L sherwood oil (boiling point 60-90 ℃), vortex 1 minute, standing demix takes off a layer methanol-water 3m L, add 14mL water, cross 0.22 μ m organic film, get 8mL filtrate and 5mg graphene oxide mixed at room temperature, concussion 30min.Centrifugal 15 minutes of 13000 rpm, abandoning supernatant adds 2ml methyl alcohol, and mixing is centrifugal, and collect supernatant, and in the aforementioned centrifugal precipitation that obtains, add methyl alcohol, mixing, centrifugal, this supernatant that obtains and aforementioned supernatant are merged, namely contain AFB
1Meoh eluate, the methyl alcohol that nitrogen blows to the meoh eluate is closely dried, add 100 μ L trifluoroacetic acids and 200 μ L normal hexanes, vortex mixing 30 seconds, 40 ℃ derive 20 minutes, and nitrogen blows organic phase, add 1mL 15% acetonitrile water, mixing, through the peak area of liquid chromatogram measuring aflatoxin, quantitative by typical curve.The recovery is tried to achieve by the actual value that records and the known ratio that adds scale value.Testing result repeats through 3 times, tries to achieve the coefficient of variation.
As a comparison experiment is got the same peanut sample of 5g and is added 15ml 70% methanol-water, vortex mixing, 50 ℃ ultrasonic 12 minutes, quantitative filter paper filters, and gets filtrate 4mL and adds 2m L sherwood oil (boiling point 60-90), vortex 1 minute, standing demix takes off a layer methanol-water 3m L, adds 8mL water, cross 0.22 μ m organic film, get 8mL filtrate and cross immune affinity column, use the 1mL methanol-eluted fractions, nitrogen blows meoh eluate to be done near, add 100 μ L trifluoroacetic acids and 200 μ L normal hexanes, vortex mixing 30 seconds, 40 ℃ derive 20 minutes, and nitrogen blows organic phase, add 1mL 15% acetonitrile water, mixing, through the peak area of liquid chromatogram measuring aflatoxin, quantitative by typical curve.The recovery is tried to achieve by the actual value that records and the known ratio that adds scale value.Testing result repeats through 3 times, tries to achieve the coefficient of variation.
The recovery of two methods and cost contrast are as follows:
| Method | The recovery (the %) ± coefficient of variation (%) | Cost (unit/sample) |
| The affine method of immunity | 89.2±8.8 | 120-180 |
| This experimental technique | 94.5±12.3 | 40-60 |
Claims (9)
1. AFB
1The liquid chromatographic detection enrichment method is characterized in that: it may further comprise the steps:
(1) AFB
1The preparation of enrichment material:
In crystalline flake graphite, add nitration mixture and potassium permanganate that sulfuric acid and phosphoric acid form, carry out preliminary oxidation reaction, then form the graphite oxide that expands by hydrolysis reaction, repeatedly wash successively purifying with hydrogen peroxide, hydrochloric acid and deionized water and obtain graphite oxide afterwards, then with the graphite oxide behind the purifying by the high-intensity ultrasonic mono-layer graphite oxide alkene that is dissociated into, obtain through the centrifugal 2000rpm of choosing not sedimentation in centrifugal 30 minutes and in the centrifugal 30min sedimentation of 5000rpm again;
(2) with the AFB of above-mentioned preparation
1Enrichment material adds in the methanol aqueous solution of testing sample, the content of methyl alcohol the highest 12.5% of the system cumulative volume that do not exceed of percentage meter by volume in the described system, described AFB
1The quality of enrichment material will reach 0.625g/mL with the system cumulative volume than at least, mixed at room temperature, and 10-30min is to carry out AFB in concussion
1Enrichment, centrifugal, remove supernatant, then carry out wash-out with methyl alcohol, with AFB
1The AFB of enrichment in the enrichment material
1Be eluted in the methyl alcohol, obtain containing AFB
1Meoh eluate, liquid chromatographic detection is for subsequent use.
2. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: the system of described step (2) adds water when needed dilutes, so that methanol content the highest 12.5% of the system cumulative volume that do not exceed of percentage meter by volume in the system; The methanol aqueous solution consumption of described testing sample is 4-8mL.
3. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: AFB in the described step (2)
1The quality of enrichment material is 0.625-0.75g/mL with system cumulative volume ratio.
4. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: AFB in described step (2) system
1Content should be in the 0.8-100ng/mL scope, as exceed, can carry out the methanol aqueous solution of testing sample carrying out again enrichment after the corresponding concentrated or dilution process.
5. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: the methanol-eluted fractions step is in the described step (2): with the methyl alcohol AFB that joined enrichment
1AFB
1In the enrichment material, mixing, centrifugal, collect supernatant, be and contain AFB
1Meoh eluate.
6. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: the methanol-eluted fractions step also comprises the repetition elution step in the described step (2), namely adds methyl alcohol in the aforementioned centrifugal precipitation that obtains, mixing, centrifugal, supernatant and the aforementioned supernatant that obtains merged, obtain containing AFB
1Meoh eluate.
7. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: described step (2) also comprises blows the methyl alcohol in the meoh eluate near doing, and adds trifluoroacetic acid and normal hexane and derives, and then adds mobile phase and carries out constant volume, for subsequent use.
8. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: described AFB
1The mass ratio of potassium permanganate and crystalline flake graphite is 6:1 in the preparation of enrichment material; The volume ratio of sulfuric acid and phosphoric acid is 9:1; The massfraction of described graphite in preliminary oxidizing process is 0.70% ~ 0.75%, and the massfraction in the end reaction system after hydrolysis is 0.3% ~ 0.375%; Described preliminary oxidation reaction condition is 50 ℃ of stirring reaction 12-48 hours.
9. according to claim 1 AFB
1The liquid chromatographic detection enrichment method is characterized in that: in the described step (1) ultrasonic power be 800W or more than, ultrasonic time be 1h or more than.
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| CN106770806A (en) * | 2017-03-03 | 2017-05-31 | 浙江工业大学 | It is a kind of to determine aflatoxin B1, the method for B2 contents in Chinese patent drug simultaneously |
| CN106841480A (en) * | 2017-03-06 | 2017-06-13 | 上海市农业科学院 | A kind of enrichment purification method of aflatoxin |
| CN107875737A (en) * | 2017-11-14 | 2018-04-06 | 中国农业科学院油料作物研究所 | A kind of adsorption of aflatoxin filter cloth, preparation method and applications |
| CN110711405A (en) * | 2019-09-29 | 2020-01-21 | 中国农业科学院油料作物研究所 | Synchronous enrichment and purification method of aflatoxin B1 and capsaicin |
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