CN107478478A - A kind of fumonisin B1Substrate standard substance and preparation method thereof - Google Patents

A kind of fumonisin B1Substrate standard substance and preparation method thereof Download PDF

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CN107478478A
CN107478478A CN201710671618.4A CN201710671618A CN107478478A CN 107478478 A CN107478478 A CN 107478478A CN 201710671618 A CN201710671618 A CN 201710671618A CN 107478478 A CN107478478 A CN 107478478A
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standard substance
substrate standard
preparation
agricultural product
substrate
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韩铮
赵志辉
郭文博
聂冬霞
何卫警
杨俊花
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Shanghai Academy of Agricultural Sciences
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Shanghai Academy of Agricultural Sciences
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/38—Diluting, dispersing or mixing samples
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N2001/2893—Preparing calibration standards

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
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  • General Health & Medical Sciences (AREA)
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Abstract

The invention provides a kind of FB1The preparation method of substrate standard substance, this method comprise the following steps：By FB1Toxigenic fungi inoculation into the culture medium of edible agricultural product, culture production poison, high toxin producing medium is freezed using freeze drier, pulverized and sieved；According to proportional diluted principle, adding the edible agricultural product powder of blank makes FB1To magnitude level, uniform FB is remixed1Edible agricultural product substrate standard substance.The FB that the present invention is prepared1Edible agricultural product substrate standard substance, be daily FB1Residue detection provides magnitude tracing material, solves the problems, such as the testing result inaccuracy caused by matrix difference, accurately ensure that FB in the agricultural product such as corn, wheat and rice1Monitoring.

Description

A kind of fumonisin B1Substrate standard substance and preparation method thereof
Technical field
The present invention relates to agricultural product security detection technique field, is specifically related to a kind of fumonisin B1Matrix standard Material and preparation method thereof.
Background technology
Fumonisin is mainly by fusarium moniliforme (Fusariummoniliforme) under certain temperature and damp condition Caused water-soluble secondary metabolite, so far there occurs a kind of ten fumonisins, wherein most importantly lying prostrate horse Toxin B1(FB1).Research shows, takes in FB1Horse cerebral white matter malacosis (Equine can be caused Leucoencephalomalacia, ELEM), pig pulmonary edema syndrome (Porcine Pulmonary Edema, PPE), to old The toxicity research of mouse shows, FB1Easily liver or kidney are caused to damage.In addition, FB1Also there is very strong cytotoxicity and exempt from Epidemic disease toxicity, it is closely related with the generation of mankind's cancer of the esophagus.Therefore, IARC (International Agency For Research on Cancer, IARC) FB12B levels carcinogenic substance is classified as the potential carcinogen of the mankind.In China, FB1It is even more a kind of widely distributed, huge mycotoxin is endangered, accounts for the 70% of fumonisin total amount of pollutants discharge.
Currently used for detecting FB1Method mainly include：After ELISA, liquid chromatography tandem mass spectrometry, derivatization Liquid chromatogram connection uv detection method etc..It is same but different experiments room even same laboratory uses different detection methods Obvious difference between the analysis result of sample be present, value is difficult to accomplish unification, it is difficult to ensures different experiments room testing result Comparativity.
Substrate standard substance is a kind of measurement criteria for having high homogeneity, good stability and value accuracy, Be advantageous to overcome the error caused by matrix effect, so as to ensure accurate and value the traceability of testing result, simultaneously It can be used for calibration instrument, verify the quality control in detection method and laboratory.Further, since FB1Standard solution and quilt FB1There is larger difference in the wheat and corn sample of pollution, its physiologically active entered in humans and animals body, bioconversion road Footpath and toxicity power are inconsistent, so as to cause its metabolism, toxicological study result it is unreliable.And substrate standard substance is in property Above, matrix composition is upper consistent with actual sample, therefore, substitutes pure verticillium toxin standard solution using substrate standard substance and carries out Research in this respect has more application value and directive significance.At present, there has been no FB1Substrate standard substance and preparation method thereof Report.
The content of the invention
The purpose of the present invention, which first consists in, provides a kind of FB1The preparation method of substrate standard substance, this method include as follows Step：
By FB1Toxigenic fungi inoculation into the culture medium of edible agricultural product (such as corn, wheat, rice), culture Production poison, high toxin producing medium is freezed, pulverize and sieve using freeze drier；
According to proportional diluted principle, adding the edible agricultural product powder (such as corn, wheat, rice) of blank makes FB1To value water It is flat, remix uniform FB1Edible agricultural product (Wheat/Maize/rice) substrate standard substance；
This method can provide FB1Edible agricultural product (such as corn, wheat, rice) substrate standard substance；
Wherein toxigenic bacterium strain is FB1High yield poison fungal bacterial strain, such as fusarium moniliforme (Fusarium moniliforme)； The production poison ring border of bacterial strain：Temperature is 10-30 DEG C, humidity 0.92-0.98aw, culture medium is corn, wheat or rice, black dull Culture, incubation time are 7-35 days；
Culture medium wherein after bacterial strain production poison pulverizes and sieves (mesh of 60 mesh -500) after must being freezed by freeze drier, freezes The dry time is 0.5-7 days；
High toxin producing medium after sieving, after adding the edible agricultural product powder of blank, it is necessary to mixed by mixed plastic device equal It is even；Wherein used blender is three-dimensional mixer, what tempering tank must using the not Plastic Drum of adhesive mixture or quite Container, can not be steel drum or metal bucket etc.；
Wherein described incorporation time changes and changed with the value of the substrate standard substance of preparation, when preparation 0-100g bases During body standard substance, required incorporation time is 0.1-4h, when preparing 100-300g substrate standard substances (being free of 100g), institute The incorporation time needed is 0.5-8h, and when preparing 300-500g substrate standard substances (being free of 300g), required incorporation time is 0.5-16h, when preparing 500-2000g substrate standard substances (being free of 500g), required incorporation time is 0.5-24h；
It should be without FB that wherein used blank, which eats agricultural product powder,1Dried powder, i.e., less than detection method examine The μ g/kg of rising limit 1；
Present invention also offers the FB being prepared by above-mentioned preparation method1Substrate standard substance, the matrix reference material Matter is dispensed after uniformity testing；
Wherein described packing process is that the substrate standard substance that will be finally prepared is dispensed using brown reagent bottle ,- 20~50 DEG C of storages.
Wherein described uniformity testing is with evidence《Primary standard material technical specification》The requirement pair of (JJG 1006-1994) The data of measurement carry out variance analysis, judge FB by the comparison of between-group variance and intra-class variance1Substrate standard substance Systematic divergence is whether there is between measured value, whether criterion material is uniform.
The beneficial effects are mainly as follows：
(1) present invention firstly discloses FB1Rice, corn and wheat substrate standard substance preparation method, be day Normal FB1Residue detection provides magnitude tracing material, solves the problems, such as the testing result inaccuracy caused by matrix difference, It is accurate to ensure that FB in corn, wheat and rice1Monitoring, ensured the health of people；
(2) break trade barrier, promote international trade：The disengaging of the edible agricultural product such as rice, corn, wheat for China Mouth provides reliable detection substrate standard substance, it is ensured that status of the China in international trade, has more far-reaching social meaning Justice；
(3) FB is studied for China1Animal experiment in vivo more realistic standard items be provided supported：Due to mycotoxin Standard solution with larger difference be present by wheat, corn and the rice sample of mycotoxin pollution, it enters people and dynamic Physiologically active, bioconversion path and toxicity power in object is inconsistent, therefore, using FB1Substrate standard substance substitutes Pure FB1Standard solution carries out research in this respect, it is cost-effective simultaneously, with more actual application value and directive significance；
(4) FB of the invention1Corn, wheat and rice substrate standard substance pass through uniformity and study on the stability, examine It is satisfactory to examine result, carries out definite value and uncertainty evaluation.Evaluation result shows, FB of the invention1Corn, wheat and Uncertainty caused by rice substrate standard substance is small, and measurement result is relatively stable, it can be ensured that the accuracy of measurement result.
Brief description of the drawings
Fig. 1 is FB1The Liquid Chromatography-Tandem Mass Spectrometry figure (concentration 25ng/mL) of standard solution
Fig. 2 is the Liquid Chromatography-Tandem Mass Spectrometry figure (content 4.5mg/kg) of X2 corn substrate standard substances
Fig. 3 is the Liquid Chromatography-Tandem Mass Spectrometry figure of blank corn flour
Embodiment
The present invention is further described with reference to specific embodiment, but protection scope of the present invention is not limited to that：
Embodiment 1
1. material, instrument and reagent
1.1 material
Corn flour (detects, it is free of FB through 2.6 method1) buy in Shanghai large supermarket, fusarium moniliforme (Fusarium moniliforme) is provided by Academy of Agricultural Sciences, Shanghai City agricultural product quality standard and detection technique research institute.
FB1Standard items (50 μ g/mL) are purchased from Sigma-Aldrich companies (St.Louis, MO, USA).
1.2 culture medium
Potato dextrose agar (PDA)：40.0g PDA powder is weighed in 1L distilled water, heating boil to It is completely dissolved, 121 DEG C of autoclaving 20min are standby.
Corn culture medium：50g corn flour is weighed to 250mL triangular flasks, 20mL distilled water is added and mixes, use breathable sealing film Bottleneck is sealed, soaked overnight, autoclaving (121 DEG C, 20min), is shaken corn flour after cooling scattered stand-by.
1.3 key instrument
Waters Ultra Performance Liquid Chromatography instruments (Waters, US), the triple level Four bar mass spectrograph Warters of series connection TQS (Warters companies of the U.S.), HeraeusMultifuge X3 supercentrifuges (U.S. Thermo Fisher Scientific companies), Thermo Scientific Heraguard ECO super-clean benches (U.S. Thermo Fisher Scientific companies), mold incubator (the upper grand experimental facilities Co., Ltd of Nereid), SX-500 high-pressure sterilizing pots (Japan TOMY companies), water-bath nitrogen evaporator (Qingdao Lu Bo enviromental protection instruments company).
1.4 main agents
PDA powder (Beijing Luqiao Technology Co., Ltd.)；Methanol, acetonitrile are purchased from Merck companies of the U.S. (chromatographically pure)； Ammonium acetate is purchased from Aladdin companies of the U.S. (chromatographically pure).
2. experimental method
2.1 UHPLC-MS/MS testing conditions
Chromatographic column：Agilent Poroshell 120EC-C18(100mm×3.0mm,2.7μm)(Agilent,CA, USA)；Mobile phase：A phases are methanol solution；B phases are 5mmol ammonium acetate aqueous solutions.Gradient elution：0~0.5min, 10%A； 6min, 90%A；6.5min, 90%A；6.7min, 10%A；8min, 10%A；Flow velocity is 400 μ L/min；The μ L of sample size 3；Post 40 DEG C of temperature.
Using electron spray cation scan pattern (ESI+), desolventizing gas, taper hole gas are high pure nitrogen, and collision gas is height Pure argon, desolventizing temperature are 500 DEG C, and 150 DEG C of ion source temperature, taper hole air-flow and desolventizing gas stream are 7.0 Hes respectively 1000L/h, by multiple-reaction monitoring (MRM) pattern to target compound accurate quantitative analysis, FB1Parent ion, daughter ion, impact energy The parameters such as amount are shown in Table 1.
The FB of table 11Mass Spectrometry Conditions parameter
Note：* quota ion is represented
2.2 FB1The configuration of standard solution
The FB that will be commercially available1Standard items obtain a series of standard items by acetonitrile/water solution (50/50, v/v) dilution Solution (5ng/mL, 10ng/mL, 25ng/mL, 50ng/mL, 100ng/mL), wherein 25ng/mL standard solution test MRM Collection of illustrative plates is as shown in Figure 1.
2.3 bacterial strains activate
By fusarium moniliforme inoculation in PDA culture medium, 28 DEG C, cultivated 5 days under dark condition, it is standby.
2.4 condition of culture and method
A diameter of 0.5cm mycelia block is taken on bacterium colony with aseptic card punch, by inoculated by hypha block into corn culture medium, Totally 50 parts, sealed with sterile breathable sealing film, be placed under 28 DEG C of dark conditions and cultivate 4 weeks.It is inoculated with the 1st week, vibrates triangle daily Bottle 1 time, makes mycelia fully be contacted with culture medium.
2.5FB1It is prepared by corn powder substrate standard substance
After culture terminates, the toxin producing medium in conical flask is taken out and is placed in 50 DEG C of drying, powder is broken into and crosses 60 mesh sieves After will contain high concentration FB1Corn culture medium powder seal up for safekeeping it is stand-by in valve bag.
Weigh the above-mentioned 100g being prepared and contain high concentration FB1Corn culture medium powder and 100g blank corn flour It is put into (after testing without FB1) in Plastic Drum, covers tightly bung, Plastic Drum is put into DWH three-dimensional mixers, and (Shanghai moral gets over powder machinery Co., Ltd, Shanghai, China) on, mix 3 hours, mixing takes out Plastic Drum after terminating, and continues to weigh 200g corn flour and pours into it In, after mixing 4 hours, prepare FB1Corn powder substrate standard substance X1.
50g X1 are weighed, and add 50g corn flour thereto, mixes 2 hours and adds 100g corn flour thereto after terminating, Mix 3 hours and add 200g corn flour after terminating thereto again, mix 4 hours and add 400g corn flour after terminating thereto again, 100g mixed powders are taken out in another Plastic Drum from the Plastic Drum after mixing 5 hours, are added 100g corn flour thereto, are mixed 3 Hour adds 200g corn flour thereto again after terminating, and mixes 4 hours and adds 400g corn flour after terminating thereto again, mixes 5 FB is prepared after hour1Corn powder substrate standard substance X2.
100g X2 are weighed, add 100g corn flour thereto, add 200g corn flour after mixing 2 hours thereto again, are mixed Prepare FB within even 3 hours1Corn powder substrate standard substance X3.
2.6 FB prepared1The detection method of FB1 contents in agricultural product substrate standard substance and blank sample
2.00 ± 0.01g measuring samples accurately are weighed, are placed in 50mL centrifuge tubes, add 10.0mL acetonitrile-aqueous solutions (50/50, v/v) extractant is used as, vortex mixed 30s, soaks 5min, ultrasonic extraction 1h, 4000r/min centrifugation 10min.Take Clear liquid crosses 0.22 μm of filter membrane, and UHPLC-MS/MS methods are used after diluting 10000 times with methanol-water solution (20/80, v/v) (2.1) detect.
3. the FB of different content1The preparation and checking of substrate standard substance
FB in 3.1 substrate standard substances1Quantitative determination
FB is carried out using 2.6 method1Quantitative detection, the results showed that, FB1Corn substrate standard substance X1 in FB1Content For 451.05mg/kg, FB in substrate standard substance X21Content is 4.19mg/kg, FB in substrate standard substance X31Content be 1.07mg/kg.Wherein substrate standard substance X2 MRM collection of illustrative plates is as shown in Figure 2.Compared with blank corn flour (accompanying drawing 3), prepare The substrate standard substance X2 of gained has obvious FB1FB in chromatographic peak, with standard solution1Chromatographic peak (accompanying drawing 1) unanimously, table The bright substrate standard substance prepared is FB1Substrate standard substance.3.2FB1The uniformity testing of substrate standard substance
According to《Primary standard material technical specification》The uniformity sampling of (JJG 1006-1994) requires, from the FB of this batch1Base 15 bottles are randomly selected in body standard substance respectively, to each sample replication 3 times.Using method of analysis of variance (with specific reference to ISO Directive/guide 35《The rule and statistical method of standard substance/standard sample definite value》) to FB1Substrate standard substance carries out uniformity Examine.
Between-group variance：
Intra-class variance：
F<Fα(v1,v2), show that no significant difference, sample are uniform between group in group；Otherwise sample is uneven.
3.3 FB1The short-term stability of substrate standard substance is examined
The FB that will be prepared1Substrate standard substance is respectively placed at 4 DEG C, -20 DEG C, 50 DEG C, investigates FB in 8 days1Matrix The stability of standard substance, wherein 50 DEG C are to simulate limit traffic condition to investigate FB1The stability of substrate standard substance.
3.4 FB1The long-time stability of substrate standard substance are examined
Tri- batches of FB1 corn powder substrate standard substances of X1, X2, X3 are taken, are divided in clean black plastic bottle, per bottled About 25g, it is individually positioned in 4 DEG C, 1~6 month in -20 DEG C of environment, investigates its stability.
4. result and analysis
Various concentrations FB in 4.1 corn flour1The uniformity of substrate standard substance
The uniformity data of tri- batches of FB1 corn substrate standard substances of X1, X2, X3 is shown in Table 2-4.According to the free degree (14,30) And given level of significance α=0.05, critical F can be checked in by F tablesα=2.04.FB1The F of tri- kinds of contents levels of X1, X2, X3 Value is respectively less than 2.04 i.e. Fx<Fα, then it is assumed that the interior no significant difference, wherein S between group of group1 2Represent between-group variance, S2 2In representative group Variance, the results showed that the horizontal FB of the different content of prepared acquisition1The uniform sexual satisfaction requirement of substrate standard substance.
The FB of table 21Substrate standard substance X1 uniformity (mg/kg)
The FB of table 31Substrate standard substance X2 uniformity (mg/kg)
The FB of table 41Substrate standard substance X3 uniformity (mg/kg)
FB in 4.2 corn flour1The short-term stability of substrate standard substance
Tri- batches of FB1 corn flour substrate standard substances of X1, X2, X3, short-term stability evaluation are carried out at different temperatures, each 3 parts of concentration is parallel, places 2 days, 4 days, 6 days, 8 days respectively, the FB surveyed under -20 DEG C, 4 DEG C and 50 DEG C temperature conditionss1Changes of contents, Its short-term stability is investigated, specific data are shown in Table 5；Calculated with line fitting approach and evaluate whether it stablizes, FB1- 20 DEG C, 4 DEG C and 50 DEG C at the short-term stability evaluation result deposited be shown in Table 6.When confidence level is 0.95, t distribution Significance factors are 3.182.Compare | b1| with t0.95, n-2×s(b1) value, if | b1| ＜ t0.95, n-2×s(b1), illustrate stability test result straight line Slope be inapparent, then show in the time range (8 days) in investigation, the sample has good stability, otherwise for It is unstable.
FB in the corn flour of table 51The short-term stability result (mg/kg) of substrate standard substance
FB in the corn flour of table 61Short-term stability evaluation result of the substrate standard substance under condition of different temperatures
Note：t0.95,3=3.182
As a result show, FB under concentration level each at each temperature1Substrate standard substance FB in 8 days1Content can keep steady Fixed, nothing significantly rises or falls trend.
4.3FB1The long-time stability of substrate standard substance
Tri- batches of FB1 corn flour substrate standard substances of X1, X2, X3, long-time stability evaluation are carried out at different temperatures, each 3 parts of concentration is parallel, places 0,1,3,6,12,18,24 month respectively, the FB under -20 DEG C of survey, 4 DEG C of temperature conditionss1Changes of contents, Its long-time stability is investigated, specific data are shown in Table 7；Calculated with line fitting approach and evaluate whether it stablizes, FB1- 20 DEG C, 4 The long-time stability evaluation result deposited at DEG C is shown in Table 8.
FB in the corn flour of table 71The long-time stability result (mg/kg) of substrate standard substance
FB in the corn flour of table 81Long-time stability evaluation result of the substrate standard substance under condition of different temperatures
Note：t0.95,3=3.182
As a result show, FB in corn flour under concentration level each at each temperature1Substrate standard substance is stable in 24 months Property it is good, without significantly rising or falling trend.In view of actual shipment and the needs of storage condition, our 4 DEG C of final choices For FB in corn flour1Substrate standard substance transports in short term and long-term storage condition.
4.4 FB1The definite value and analysis on Uncertainty of substrate standard substance
Organize Technical Center for Animal, Plant and Food Inspection and Quarantine, Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai City animal epidemic pre- Anti- control centre, Shanghai Bi Nuo detection techniques Services Co., Ltd, Expert Testimony Science-Technology Inst., Judical Department, Chinese agriculture 6 units such as academy of sciences's agricultural product quality standard and research institute, Shanghai Disease Prevention and Control Centre are to above three concentration FB1Substrate standard substance (X1, X2 and X3) has carried out definite value experiment, finally gives FB in corn flour substrate standard substance1Content Respectively 451.05mg/kg, 4.19mg/kg, 1.07mg/kg, relative standard deviation are followed successively by 0.25%, 0.44,1.11%.
The substrate standard substance definite value product test of table 9
4.5 FB1The uncertainty evaluation of substrate standard substance
FB1The uncertainty of substrate standard substance is made up of three parts：The uncertainty that the purity of standard substance introduces (uc)；Uncertainty that more unit measures introduce (including the preparation of standard liquid during measurement analysis, sample weighing, The uncertainty caused by influence factor such as instrument detection and the rate of recovery) (uchar)；Uniformity (ubb) and stability (usts,ulst) The uncertainty of introducing.
Each partial uncertainty such as table 10, X1, X2, X3FB1Definite value and uncertain knot in corn flour substrate standard substance Fruit is respectively：451.05±24.94mg/kg、4.19±0.24mg/kg、1.07±0.11mg/kg.
FB in the corn flour of table 101Standard substance content definite value and uncertainty
As a result：
(1) toxin producing medium contains a large amount of FB after measured1, the FB of a variety of concentration can be met1The preparation of substrate standard substance will Ask, make it have wider array of application；
(2) Analysis of variance is examined, and the standard substance sample homogeneity being prepared through this method is good, meets《One-level Standard substance technical specification》(JJG 1006-1994) standard substance demand of technical standard；
As fully visible, the FB in a variety of agricultural product such as corn, wheat, rice can be effectively prepared in the present invention1Base Body standard substance, product have good uniformity, will be generally investigated for agricultural products in China quality safety and Risk Monitoring provides value Trace to the source and technical support.
Protection scope of the present invention is not limited to the description made in embodiment, without departing from the modification at the present invention program center Belong to protection scope of the present invention.

Claims (5)

  1. A kind of 1. FB1The preparation method of substrate standard substance, it is characterised in that this method comprises the following steps：
    By FB1Toxigenic fungi inoculation into the culture medium of edible agricultural product, culture production poison, using freeze drier by height Toxin producing medium freezes, and pulverizes and sieves；
    According to proportional diluted principle, adding the edible agricultural product powder of blank makes FB1Mixed to magnitude level, then by mixed plastic device It is uniform to obtain FB1Edible agricultural product (Wheat/Maize/rice) substrate standard substance.
  2. 2. FB according to claim 11The preparation method of substrate standard substance, it is characterised in that
    Wherein toxigenic bacterium strain is FB1High yield poison fungal bacterial strain fusarium moniliforme (Fusarium moniliforme)；
    The production poison ring border of wherein bacterial strain is：Temperature is 10-30 DEG C, humidity 0.92-0.98aw, black dull culture, incubation time 7- 35 days.
  3. 3. FB according to claim 11The preparation method of substrate standard substance, it is characterised in that
    Culture medium wherein after bacterial strain production poison by freeze drier it is lyophilized after pulverize and sieve the mesh of 60 mesh -500, freeze-drying time is 0.5-7 days.
  4. 4. FB according to claim 11The preparation method of substrate standard substance, it is characterised in that
    Wherein described incorporation time changes and changed with the value of the substrate standard substance of preparation, when preparation 0-100g matrix marks During quasi- material, required incorporation time is 0.1-4h, and when preparing 100-300g substrate standard substances, required incorporation time is 0.5-8h, when preparing 300-500g substrate standard substances, required incorporation time is 0.5-16h, when preparation 500-2000g bases During body standard substance, required incorporation time is 0.5-24h.
  5. A kind of 5. FB being prepared by any one of claim 1-4 methods described1Substrate standard substance.
CN201710671618.4A 2017-08-08 2017-08-08 A kind of fumonisin B1Substrate standard substance and preparation method thereof Pending CN107478478A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109490028A (en) * 2018-08-27 2019-03-19 江苏省疾病预防控制中心 Inorganic arsenic standard sample preparation method in a kind of rice meal
CN109503393A (en) * 2018-11-07 2019-03-22 江苏省农业科学院 A kind of high speed adverse current chromatogram prepares fumonisin B1The method of standard items
CN109900533A (en) * 2019-02-26 2019-06-18 齐鲁师范学院 A kind of home position observation method of plant root hair
CN118168885A (en) * 2024-05-15 2024-06-11 中检科(北京)测试认证有限公司 Rice bran powder matrix standard sample containing fumonisin B1 and preparation method thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102288464A (en) * 2011-07-05 2011-12-21 时文春 Preparation method for standard sample of chloramphenicol residual lyophiled powder in muscle of carp
CN104596824A (en) * 2015-02-14 2015-05-06 福建出入境检验检疫局检验检疫技术中心 Chicken matrix reference material containing amantadine and ribavirin and preparation method of chicken matrix reference material
CN104931308A (en) * 2015-05-18 2015-09-23 上海市农业科学院 Method for simultaneously preparing standards of fumonisins B1, B2 and B3
CN105547788A (en) * 2016-02-23 2016-05-04 蒋丹 Fusarium moniliforme qualitative and standard sample in bean flour and preparation method thereof
CN106813963A (en) * 2016-04-06 2017-06-09 中国计量科学研究院 A kind of fish powder standard substance preparation method containing chloramphenicol

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102288464A (en) * 2011-07-05 2011-12-21 时文春 Preparation method for standard sample of chloramphenicol residual lyophiled powder in muscle of carp
CN104596824A (en) * 2015-02-14 2015-05-06 福建出入境检验检疫局检验检疫技术中心 Chicken matrix reference material containing amantadine and ribavirin and preparation method of chicken matrix reference material
CN104931308A (en) * 2015-05-18 2015-09-23 上海市农业科学院 Method for simultaneously preparing standards of fumonisins B1, B2 and B3
CN105547788A (en) * 2016-02-23 2016-05-04 蒋丹 Fusarium moniliforme qualitative and standard sample in bean flour and preparation method thereof
CN106813963A (en) * 2016-04-06 2017-06-09 中国计量科学研究院 A kind of fish powder standard substance preparation method containing chloramphenicol

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
NORHAFNIZA AWALUDIN ET AL.: "Preparation of an In-House Reference Material Containing Fumonisins in Thai Rice and Matrix Extension of the Analytical Method for Japanese Rice", 《TOXINS》 *
董莲华 等: "转基因玉米NK603基体标准物质研制", 《农业生物技术学报》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109490028A (en) * 2018-08-27 2019-03-19 江苏省疾病预防控制中心 Inorganic arsenic standard sample preparation method in a kind of rice meal
CN109503393A (en) * 2018-11-07 2019-03-22 江苏省农业科学院 A kind of high speed adverse current chromatogram prepares fumonisin B1The method of standard items
CN109503393B (en) * 2018-11-07 2021-10-26 江苏省农业科学院 Preparation of fumonisins B by high-speed countercurrent chromatography1Method for preparing standard substance
CN109900533A (en) * 2019-02-26 2019-06-18 齐鲁师范学院 A kind of home position observation method of plant root hair
CN118168885A (en) * 2024-05-15 2024-06-11 中检科(北京)测试认证有限公司 Rice bran powder matrix standard sample containing fumonisin B1 and preparation method thereof
CN118168885B (en) * 2024-05-15 2024-08-06 中检科(北京)测试认证有限公司 Rice bran powder matrix standard sample containing fumonisin B1 and preparation method thereof

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Application publication date: 20171215