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Mercapturic acid adduct detection method for evaluating short-term exposure of acrylamide and application

A technology of mercaptouric acid and acrylamide, which is applied in the field of simultaneous detection of mercaptouric acid adducts, can solve the problem of unfavorable large-scale biological sample detection, low-content sample detection, simultaneous quantitative analysis of four mercaptouric acid adducts, and long analysis time. And other issues

Inactive Publication Date: 2015-12-16
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current quantitative analysis method does not achieve simultaneous quantitative analysis of the four mercaptouric acid adducts
At the same time, the above-mentioned methods such as HPLC take a long time to analyze, and the detection sensitivity is very limited, which is not conducive to the detection of large quantities of biological samples and the detection of low-content samples

Method used

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  • Mercapturic acid adduct detection method for evaluating short-term exposure of acrylamide and application
  • Mercapturic acid adduct detection method for evaluating short-term exposure of acrylamide and application
  • Mercapturic acid adduct detection method for evaluating short-term exposure of acrylamide and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Embodiment 1. A method for synchronous detection of mercaptouric acid adducts for evaluating short-term in vivo exposure of dietary acrylamide. The detection object is rat urine, and the following steps are performed in sequence:

[0118] 1), sample pretreatment:

[0119] Rat urine pretreatment method: Thaw SpragueDawley rat urine, take 10 μL mouse urine, add 10 μL mixed isotope internal standard (D 3 -AAMA,D 3 -GAMA,D 3 -IsoGAMA, the concentration is 10 μg / mL, using the diluent as the solvent), dilute 100 times with the diluent, dilute to 1 mL, vortex for 2 min to mix, filter through a 0.22 μm microporous membrane, and then inject the sample for analysis.

[0120] The diluent is: 0.1% (volume %) formic acid aqueous solution;

[0121] 2), chromatographic conditions:

[0122] Flow rate: 0.2mL / min; column temperature: 40°C.

[0123] Table 3. Gradient elution program of optimized mobile phase

[0124]

[0125] instruction manual:

[0126] A is 0.1% (volume %) for...

Embodiment 2

[0144] Example 2. A method for synchronous detection of mercaptouric acid adducts for evaluating short-term in vivo exposure to dietary acrylamide. The detection object is rat urine, and the following steps are performed in sequence:

[0145] 1), sample pretreatment:

[0146] Rat urine pretreatment method: thaw rat urine, take a certain amount of 5 μL mouse urine, add 10 μL mixed isotope internal standard (D 3 -AAMA,D 3 -GAMA,D 3 -IsoGAMA, the concentration is 10 μg / mL, using the diluent as the solvent), dilute 200 times with the diluent, set the volume to 1 mL, vortex for 2 min to mix, filter through a 0.22 μm microporous membrane, and then inject the sample for analysis.

[0147] The diluent is: 0.5% (volume %) formic acid aqueous solution;

[0148] 2), chromatographic conditions:

[0149] Flow rate: 0.2mL / min; column temperature: 40°C.

[0150] Gradient elution program of mobile phase in Table 5

[0151]

[0152] instruction manual:

[0153] A is 0.5% (volume %) f...

Embodiment 3

[0165] Example 3. A method for synchronous detection of mercaptouric acid adducts for evaluating short-term in vivo exposure to dietary acrylamide. The detection object is human urine, and the following steps are performed in sequence:

[0166] 1), sample pretreatment:

[0167] Thaw human urine, take 1mL, add 20μL mixed isotope internal standard (D 3 -AAMA,D 3 -GAMA,D 3 - Different GAMA, D 3 -AAMA sulfoxide, the concentration is 10 μg / mL, and the diluent is used as the solvent), 10 μL of formic acid and 2 mL of ammonium formate (50 mmol / L, pH 2.5) were added in sequence, vortexed for 30 s, and then centrifuged at 10000 r / min for 5 min. Then load the supernatant to a solid-phase extraction column pre-activated with 3mL of methanol, 1.5mL of water and 1.5mL of 0.1% formic acid in water, rinse with 2mL eluent after draining, remove impurities and discard the eluent solution; finally eluted with 2 mL of 1% formic acid methanol solution (that is, the volume concentration of for...

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Abstract

The invention discloses a mercapturic acid adduct detection method for evaluating short-term exposure of acrylamide. The method includes the steps that 1, urine to be detected is pretreated; 2, the pretreated urine is subjected to chromatographic detection; 3, a standard curve method is adopted for performing quantitative analysis on the detection result. An isotope dilution ultrahigh performance liquid chromatography-tandem mass spectrometry method (UHPLC-MS / MS) is adopted for quantifying four mercapturic acid adducts of acrylamide, analysis time is greatly shortened, meanwhile, the four mercapturic acid metabolins are well separated, synchronous detection of the four mercapturic acid adducts is achieved, and therefore the internal exposure quantity of acrylamide can be evaluated more comprehensively.

Description

technical field [0001] The invention relates to a method for synchronous detection of mercaptouric acid adducts for evaluating short-term in vivo exposure of dietary acrylamide and an application thereof, belonging to the field of food safety. Background technique [0002] Acrylamide (Acrylamide) is a colorless, odorless white crystalline small molecular substance, and is an important chemical raw material for the synthesis of polymers and gelling agents. The International Agency for Research on Cancer (IARC) lists acrylamide as a Class 2A carcinogen (probably carcinogenic to humans), which has neurotoxicity, reproductive toxicity, and genotoxicity. In 2002, researchers from the Swedish National Food Administration and Stockholm University jointly announced that high levels of acrylamide were found in fried foods and baked foods (Swedish National Food Administration, Information about acrylamide in food, 2002, 4). Subsequent studies have proved that the formation of acrylam...

Claims

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Application Information

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IPC IPC(8): G01N30/88
Inventor 章宇王桥程军
Owner ZHEJIANG UNIV
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