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Table 2 FDA Recommended Limits for Fumonisin in Animal Feed (June 2000)
Flow phase : methanol-0.1mol/L sodium dihydrogen phosphate solution 77:23
Flow rate : 1 mL/min      Â
Injection volume: 20μL
Fluorescence detector : λ-excitation wavelength: 335 nm λ-emission wavelength: 440 nm
Solution for mycotoxin detection in Puribond feed
First, the origin and characteristics of fumonisin
Fumonisins are mainly secondary metabolites produced by the propagation of Fusarium oxysporum f. moniliforme and f. proliferatum under certain temperature and humidity conditions. So far, the fumonisins have been found to have 11 kinds of FA1, FA2, FB1, FB2, FB3, FB4, FC1, FC2, FC3, FC4 and FP1. Grain is susceptible to contamination by the above two fungi during processing, storage and transportation, especially when the temperature is suitable, which is more conducive to its growth and reproduction, thus producing a class of toxins with similar structural properties, of which FB1 is the main component of 60. Above %, its toxicity is also the strongest. Therefore, fumonisin can cause serious damage to animal husbandry and even human health through processes such as grain processing and feed production.
The FB1 food contamination situation is widespread around the world, mainly polluting corn and corn products, and its contaminated feed is mainly corn-based feed. In 1996, China investigated the pollution of FB1 in corn, wheat and other food crops. It was found that different areas have different degrees of pollution. The contamination rate of corn fumonisin in Linxian County, a high incidence area for esophageal cancer in China, is 48%. Therefore, it is suspected that the high incidence of esophageal cancer in this area is associated with the consumption of this toxin corn. The toxin has been listed by the World Health Organization as one of several mycotoxins that were first studied in recent years.
Second, the harm of fumonisin
In 2001, the US Food and Drug Administration (FDA) issued a maximum circular guidance for fumonisin in corn and corn products for human consumption, stipulating that the maximum limit of fumonisin in human corn is 2 mg/kg; The Medical Center (CVM) has also issued a maximum amount of guidance on the regulation of fumonisin in animal feed, with a limit of 1 to 50 mg/kg.
Corn and its by-products are used in the following animal feeds
Recommended limit standard (FB1+FB2+FB3), mg/kg
Horse and rabbit
5ppm (not exceeding 20% ​​of the daily food)*
Pig and squid
20ppm (not exceeding 20% ​​of the daily food)*
Produced ruminants, poultry, crickets
30ppm (not exceeding 20% ​​of the daily ration)*
Ruminants used for slaughter for more than 3 months, used to make suede
60ppm (not exceeding 20% ​​of the daily ration)*
Poultry for slaughter
100ppm (not exceeding 20% ​​of the daily ration)*
Various other animals and pets
10ppm (not exceeding 20% ​​of the daily ration)*
*Based on dry basis
Fourth, the detection of fumonisin
Immunoaffinity column + fluorometric assay and HPLC method.
In line with the national standard GBT 25228-2010 grain and oil testing corn and its products in the determination of fumonic acid content immunoaffinity column purification high performance liquid chromatography and fluorescence spectrophotometry.
V. Introduction of Puriban Fumonisin Detection Program
(1) Immunoaffinity column - high performance liquid chromatography:
Pribolab® uses immunoaffinity column purification, and uses HPLC and fluorescence detectors to detect HPLC assays that provide fumonisin assays. The results are accurate and reliable, and the detection limits are good. A good method for detecting fumonisin.
1. Equipment and consumables configuration
Serial number
Product name
Specifications and parameters
1
High performance liquid chromatography and fluorescence detector
2
Pribolab mycotoxin-specific column
150/250mm Item No.: PRC-18
3
PriboFast® Fumonisin Immunoaffinity Column
3ml, 25pcs/box, article number: IAC-050-3
4
High speed homogenizer
Up to 22,000 rpm or more
5
PriboFast® fiberglass filter paper
100P, 110mm, 1.5μm Item No.: GMF-110
6
PriboFast® eight-position pump flow operator
Item No.: EQ-PUMP-8
7
Fumonisin standard
Solid or liquid
8
Derivatization reagent
2-mercaptoethanol MCH2-Mercaptoethanol
O-phthalaldehyde
2. Sample preparation: Puribang offers different treatment options for foodstuffs
Peanuts, corn, rice, wheat and its products and feed
- Place 50 g of ground sample + 5 g of salt in a homogenized cup.
- Add 100 mL of methanol: water (80:20) solution.
---- Cover the lid and mix at high speed for 5 minutes.
Centrifuge at 4000 rpm for 5 min or filter with fluted filter paper;
---- Take 10 mL of filtrate and add 40 mL of PBS solution to dilute the filtrate and mix.
, filtered with glass microfiber filter paper , and the diluted liquid is to be tested;
The supernatant was filtered through a microfiber filter paper, and the filtrate was collected in a glass syringe barrel and weighed 10 mL.
3. Immunoaffinity column purification : enrichment-washing-elution--collect all eluents for chemical derivatization detection.
4. derivatization reaction
The above purified sample was derivatized with a mixture of o-phthalaldehyde OPA and 2 -thioethanol MCE and injected rapidly .
5. High performance liquid chromatography analysis
HPLC-column : C-18 column 150 x 4.6 mm;
Column temperature : room temperature
HPLC configuration: high performance liquid chromatography with fluorescence detector
Sixth, warm TIPS:
1. The favorable conditions for the growth and reproduction of fungi in cereals and feeds are mainly suitable temperature and moisture. If the grain, feed, etc. can be stored below 10 ° C and the water content is kept below 10%, the mold can be effectively prevented.
2. Personnel engaged in the research and testing of mycotoxins must pay attention to protection. For example, wear a protective coat cap. When performing fungal separation and culture work, wear a mask and try to prevent spores from flying.
3. If the operating table is leaking, it should be disinfected immediately with the new 5% sodium hypochlorite. When treated with 5% sodium hypochlorite (NaOCl), aflatoxin is destroyed within a few seconds and is a commonly used disinfectant.
4. There are also reports on the application of biological methods to detoxification. The low cost and high efficiency of biological methods may be a promising detoxification measure.
In view of the harm of mycotoxins to the human body, teachers who are struggling to fight on the front line of anti-drug must pay attention to protect themselves!