Method for detecting ochratoxin A (OTA) based on luminescence resonance energy transfer between up-conversion luminescence nano-material and gold nano-rods

A resonance energy transfer, ochratoxin technology, applied in the fields of nanomaterials and analytical chemistry, can solve the problems of antibodies being easily affected by external conditions, high detection cost, cumbersome and time-consuming, etc., and achieves easy chemical modification, improved sensitivity, and good stability. Effect

Active Publication Date: 2016-03-02
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, antibodies are easily affected by external conditions, especially temperature, and the preparation of antibodies requir

Method used

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  • Method for detecting ochratoxin A (OTA) based on luminescence resonance energy transfer between up-conversion luminescence nano-material and gold nano-rods
  • Method for detecting ochratoxin A (OTA) based on luminescence resonance energy transfer between up-conversion luminescence nano-material and gold nano-rods
  • Method for detecting ochratoxin A (OTA) based on luminescence resonance energy transfer between up-conversion luminescence nano-material and gold nano-rods

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Establishment of standard curve for detection of ochratoxin A in actual beer samples and pretreatment of test samples: beer samples were refrigerated at 4° C. for 30 minutes, and degassed by ultrasound. Take 20g of degassed beer sample, put it in a 25mL volumetric flask, add 2% sodium bicarbonate and 15% sodium chloride and mix the extract to the mark, mix well, filter with glass fiber filter paper until clear, collect the filtrate for later use.

[0031] 5 different types of beer were purchased from a local supermarket, and the content of ochratoxin A was determined by using the method of the present invention and the enzyme-linked immunosorbent method, and the results are shown in Table 1. The detection results of the two methods were consistent with no significant difference.

[0032] Table 1: The actual sample detection of beer, the method of the present invention is compared with the Elisa method

[0033]

[0034] Note: ND is not detected

Embodiment 2

[0035] Example 2: The detection of ochratoxin A in the actual beer sample and the recovery rate of standard addition The experimental sample pretreatment is the same as that in Example 1.

[0036] Using the 5 groups of ochratoxin A concentration data obtained in Example 1 as the background value, five OTA standard substances with different concentrations were added thereto, and the OTA content was detected again by the method of the present invention to obtain the detected value. Recovery %=(detection value-background value) / addition amount X100%. It can be seen from the data in Table 2 that the recovery rate is 93.4% to 119%, indicating that the present invention is stable, sensitive and accurate, and is applicable to the detection of OTA in actual beer samples.

[0037] Table 2: Detection and recovery of ochratoxin A in actual beer samples

[0038]

Embodiment 3

[0039] Embodiment 3: Ochratoxin A detection standard curve establishment and detection sample pretreatment in wheat actual sample

[0040] Grind wheat, hard grain, etc. with a high-speed universal grinder and pass through a 1mm aperture test sieve, do not grind into powder. Weigh 20g (accurate to 0.01g) of ground wheat sample into a 100mL volumetric flask, add 5g of sodium chloride, add 80% methanol extract to the mark, mix well, transfer to a homogeneous cup, and extract with high-speed stirring for 2min . Quantitative filter paper filtration, pipette 10.0mL filtrate into a 50mL volumetric flask, add water to volume, mix well, filter with glass fiber filter paper until the filtrate is clear, collect filtrate A in a clean container.

[0041] Nine different kinds of wheat were purchased from a local supermarket, and the content of ochratoxin A was determined by using the method of the present invention and the enzyme-linked immunosorbent method, and the results are shown in Ta...

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Abstract

The invention discloses a method for detecting ochratoxin A (OTA) based on luminescence resonance energy transfer between an up-conversion luminescence nano-material and gold nano-rods. The method is used for detecting the content of OTA in wheat and products thereof. The up-conversion luminescence nano-material (NaYF4:Yb0.286, Er0.0286) and OTA aptamers are connected to form an energy donor probe, then the energy donor probe and an energy receptor probe form a nano-composite based on the base complementary pairing principle, the phenomenon of luminescence resonance energy transfer (LRET) occurs, and the purpose of up-conversion luminescence quenching is achieved, wherein the energy receptor probe is formed by the gold nano-rods modified by aptamer complementary oligonucleotide single strands, and the aspect ratio of the gold nano-rods is about 2.5. When OTA exists in a detection system, OTA and OTA aptamers are specifically bound, and therefore double strands are melted; OTA can be quantitatively detected by monitoring the up-conversion luminescence signal intensity at 657 nm, the linearity range is 0.05-100 ng/mL, and the detection limit is 0.027 ng/mL. The method has the advantages of being high in sensitivity, fast, simple and convenient to implement when used for detecting OTA. In addition, the method is applied to detecting beer or wheat samples, and results are accurate and reliable.

Description

technical field [0001] The invention discloses a method for detecting ochratoxin A based on luminescence resonance energy transfer between upconversion luminescent nanomaterials and gold nanorods, relates to the technical field of nanomaterials and analytical chemistry, and is used for detecting ochratoxin A in food. Background technique [0002] Ochratoxins (Ochratoxins, OT) are toxic metabolites produced by certain species of Aspergillus and Penicillium, and are a family of compounds with similar molecular structures linked to isocoumarin and L-phenylalanine. Ochratoxin (OT) mainly includes seven types of ochratoxin A (OTA), ochratoxin B (OTB), ochratoxin C (OTC), and ochratoxin D (OTD), among which OTA is the main one. The most toxic, the most serious pollution, the most widely distributed. OTA is mainly produced by three fungi, Penicillium Verrucosum, Aspergillus achraceus and A. carbonarius. It mainly endangers the kidneys of humans and animals. Animal experiments hav...

Claims

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Application Information

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IPC IPC(8): G01N21/63
Inventor 吴世嘉戴邵亮王周平段诺
Owner JIANGNAN UNIV
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