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Novel arginine determination method based on LC-MS/MS

A determination method and technology for arginine, applied in the new LC-MS/MS-based arginine determination field, can solve the problems of harsh reaction conditions, narrow analytical quantification range, unfavorable accurate quantification, etc., and achieve non-toxic reaction, Ease of volatilization and removal for analysis

Active Publication Date: 2015-04-08
苏州圣苏新药开发有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

However, its structure is too short, and trace residues will reduce the service life of ordinary C18 analytical columns, which is not convenient for the processing and analysis of a large number of samples
[0009] Fluorenylmethyl chloroformate, AQC and OPA all use fluorescence detection, but the reaction conditions are harsh, and there is also the problem of narrow analytical quantitative range, and when hydroxyproline and hydroxylysine exist in the sample, the reaction by-products will be serious Interfering with quantification, so when analyzing plasma samples, it is necessary to carefully consider the influence of the matrix; dansyl chloride derivative products have greater interference, and the peak shape is poor, which is not conducive to accurate quantification; DNFB derivative reagents are more toxic; excessive derivative reagents are harmful to the analysis. certain influence

Method used

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Examples

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Embodiment Construction

[0022] This example is a new LC-MS / MS-based method for the determination of arginine, which uses a specific derivatization reagent and LC-MS / MS to accurately and quantitatively detect the derivatized arginine product. Specific steps are as follows:

[0023] Step 1: Accurately weigh 2.0 mg of arginine, and use 2.0 mL of ultrapure water to prepare an arginine stock solution with a final concentration of 1 mg / mL; then use a pipette to draw 10 μL of arginine stock solution and add it to 990 μL of Prepare a working solution of 10 μg / mL arginine in ultrapure water, take 5 μL of this working solution, add it to 95 μL of freshly collected SD rat blank plasma, and vortex it on the oscillator for 20 seconds to mix it evenly. At this time, the concentration of arginine in plasma is 500ng / mL;

[0024] Step 2: Add 300 μL of acetonitrile to the above plasma, shake vigorously on the shaker, use a centrifuge at room temperature, set the speed at 13000 rpm, centrifuge for 5 minutes, transfer ...

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Abstract

The invention discloses a novel arginine determination method based on LC-MS / MS. According to the method, specific derivatization reagents and LC-MS / MS are utilized for accurately and quantitatively detecting derivative arginine product, with the adoption of the method, the mass spectrum response value is improved by more than 10 times, the derivatization reaction is rapid, is complete within 10 minutes, and nontoxic and efficient effects are achieved; the product is well preserved on a universal analytical column C18, so that analysis is facilitated, the residual PEITC subjected to reaction is easily volatilized to be removed, moreover, a carbon chain is long, so that the analytical column is not damaged, the analysis range is four orders of magnitude within 100pg / mL-1microg / mL.

Description

technical field [0001] The present invention relates to a method based on the use of a specific derivatization reagent, in particular to a method for determining the concentration of arginine in plasma by LC-MS / MS after derivatization of arginine acid. Background technique [0002] Arginine (L-Arg) is one of the most functional amino acids in animal cells. It participates in the process of synthesizing cytoplasmic protein and nuclear protein, and plays an important role in various physiological processes such as cell division, protein synthesis and hormone secretion. [0003] Arginine is also a starting material for polyamine synthesis. Polyamine is an important biological regulatory substance, which is related to the biological metabolism of DNA, RNA and protein, and plays a key regulatory role in the process of cell growth cycle; it participates in the T cell immune response induced by mitogens, and plays a key role in regulating the central nervous system. It plays a ke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 高然史犇卜海之
Owner 苏州圣苏新药开发有限公司
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