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Phytic acid analysis method based on trimethylsilyl diazomethane methyl esterification

A technology of trimethylsilyl diazomethane and an analytical method, applied in the detection field, can solve problems such as no relevant reports and unclear detection, and achieve the effects of improving sensitivity, improving sensitivity and reproducibility, and simple ionic structure

Active Publication Date: 2021-02-05
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, whether the methylation of the phosphate group of trimethylsilyldiazomethane can be applied to the detection of phytic acid is still unclear, and there are no related reports

Method used

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  • Phytic acid analysis method based on trimethylsilyl diazomethane methyl esterification
  • Phytic acid analysis method based on trimethylsilyl diazomethane methyl esterification
  • Phytic acid analysis method based on trimethylsilyl diazomethane methyl esterification

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Experimental program
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Effect test

Embodiment 1

[0063] Embodiment 1: The working curve making of phytic acid detection under the condition of adding EDTA

[0064] Prepare a phytic acid standard stock solution with a concentration of 100 μg / mL (the solvent is water-methanol, the volume ratio is 1:9), take 1.0 mL of the stock solution, add 0.5 mol / L hydrochloric acid to adjust the pH value of the solution to 6.5, and add 150 μL trimethyl Silyl diazomethane was reacted at 25°C for 60 min, and 50 μL of formic acid was added to terminate the reaction to obtain a derivatization product solution; The phytic acid product solution was diluted step by step into 100ng / mL, 50ng / mL, 10ng / mL, 5ng / mL, 1ng / mL, 0.5ng / mL standard solution series, and analyzed by liquid chromatography-mass spectrometry respectively, and the peak area was used to analyze the phytic acid Concentration do work curve such as Figure 6 As shown, the correlation coefficient r 2 >0.999, the parameters related to the working curve are listed in Table 2. Reproducib...

Embodiment 2

[0068] Embodiment 2: the detection of phytic acid in beverage

[0069] Take 100 μL of a beverage, add hydrochloric acid with a concentration of 0.2 mol / L, adjust the pH value to 6.0, add 900 μL methanol, add 250 μL TMSD, react at 25 ° C for 60 minutes, and 10 μL ethylenediaminetetraacetic acid (EDTA) with a concentration of 10 mg / mL The solution was terminated by adding 50 μL of formic acid, blown with nitrogen, fixed to volume and filtered, then analyzed by liquid chromatography-mass spectrometry. Its LC-MS MRM selected ion diagram is shown in Figure 7 , Substituting the obtained peak area into the linear regression equation in Table 1 can obtain the content of phytic acid in the beverage. The content of phytic acid in the recorded beverage is 1.07 μg / mL. According to the standard recovery experiment, the recovery rate is 94.9%.

Embodiment 3

[0070] Embodiment 3: the detection of phytic acid in rice

[0071] Take 1g of a certain brand of rice after crushing, add hydrochloric acid with a concentration of 0.1mol / L, adjust the pH value to 6.5, add 10mL of methanol-water (V:V=1:9) and 10μL of ethylenediamine with a concentration of 10mg / mL Tetraacetic acid (EDTA) solution, magnetically stirred for 2 hours, centrifuged for 15 minutes; 100 μL of supernatant was taken, 900 μL of methanol was added, 300 μL of TMSD was added to react at room temperature for 90 minutes, and 50 μL of formic acid was added to terminate the reaction. LC-MS MRM selected ion diagram see Figure 8 . Substitute the obtained peak area into the linear regression equation in Table 2 to obtain the content of phytic acid in rice. The content of phytic acid in rice is 6.98mg / g, and the recovery experiment of adding standard records, the recovery rate is 118.6%.

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Abstract

The invention discloses a phytic acid analysis method based on trimethylsilyl diazomethane derivatization. The method comprises the following steps: regulating the pH value of a sample to be acidic, and adding a proper amount of trimethylsilyl diazomethane to perform methyl esterification; and carrying out nitrogen blowing and volume metering on the obtained derivatization product solution, then carrying out liquid chromatography-mass spectrometry analysis, and carrying out quantitative analysis by adopting an external standard method. According to the method, methyl esterification is adoptedto improve the stability of phytic acid, and the polarity, acidity and complexing capacity of metal ions of phytic acid are greatly reduced, so that liquid chromatography-mass spectrometry analysis ofphytic acid is realized under general conditions, and the sensitivity, selectivity and accuracy of the detection method are remarkably improved. Meanwhile, trimethylsilyl diazomethane is adopted as aderivatization reagent, the derivatization reaction is rapid, the reaction conditions are mild, the reaction operation is simple and convenient; meanwhile, the excessive derivatization reagent is convenient to remove, so that the analysis method is easy to popularize.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a phytic acid analysis method based on trimethylsilyl diazomethane methyl esterification. Background technique [0002] Phytic acid (also known as phytic acid ester, IP6) is ubiquitously present in the seeds, roots and stems of plants, and is the main storage form of phosphorus in plants. Among them, phytic acid or its salts in plant seeds account for about 1% of the weight of seeds. -2%. As a multifunctional molecule, phytic acid is involved in many very important cellular functions, such as RNA expression, DNA repair, P-glycoprotein regulation, insulin, innate immunity and many more. [0003] As the value of acid function is constantly being understood and excavated by people, phytic acid has been widely used as a metal removing agent, antioxidant, preservative and fermentation accelerator in the food field, and can be used to treat diabetes, kidney stones and other diseases...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 蒋可志俞赛赛汪嫣
Owner HANGZHOU NORMAL UNIVERSITY
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