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Fatty acid lc-ms/ms analysis method based on double derivatization technology

A LC-MS, double derivatization technology, applied in the field of analysis and detection, can solve problems such as poor ionization efficiency and poor mass spectrometry detection sensitivity, and achieve the effects of improving sensitivity, avoiding insufficient specificity, and mild derivatization conditions

Active Publication Date: 2022-06-07
BEIJING CHAOYANG HOSPITAL CAPITAL MEDICAL UNIV
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Problems solved by technology

However, due to the chemical structure characteristics of fatty acids, their ionization efficiency in the electrospray ion source is poor, making their mass spectrometric detection sensitivity poor

Method used

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  • Fatty acid lc-ms/ms analysis method based on double derivatization technology
  • Fatty acid lc-ms/ms analysis method based on double derivatization technology
  • Fatty acid lc-ms/ms analysis method based on double derivatization technology

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experiment example

[0057] 1 Prepare chemicals and experimental instruments

[0058] Acetonitrile (mass spectrometry grade) was purchased from Fisher Scientific (Pittsburgh, PA, USA). Formic acid (mass spec grade) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was prepared using a Milli-Q purification system (Bedford, MA, USA). All FA standards were purchased from Cayman Chemical Co. (Ann Arbor, MI, USA). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), 1-Hydroxybenzotriazole hydrate (HOBT), 2-(7-azabenzotriazole) Azole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIEA), 2-hydrazino-4,6-di Methylpyrimidine (DMP) and 2-hydrazinopyrimidine (DP) were both purchased from Sigma-Aldrich. SparkHolland liquid chromatography system (Spark, The Netherlands), API 5500 mass spectrometer (AB Sciex, Canada).

[0059] 2 Prepare the sample to be tested

[0060] 8-week-old male Wistar rats with a body weight of 250±5g (purchas...

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Abstract

The invention discloses a fatty acid LC-MS / MS analysis method based on double derivatization technology, comprising the following steps: 1. preparing a fatty acid mixed standard solution; 2. preparing a 2-hydrazinopyrimidine derivatization solution; 3. preparing a derivative 4. Prepare 2-hydrazino-4,6-dimethylpyrimidine derivatization solution; 5. Prepare each standard curve solution to be tested; 6. Use LC-MS / MS method to detect each standard curve to be tested solution, to establish a standard curve for various fatty acids; 7. Take the sample to be tested, add DMP derivatization solution and mix until the fatty acids are derivatized, then add the derivatization internal standard solution, mix well, take the supernatant after centrifugation, and use LC-MS / MS method to measure the type and content of fatty acids. The invention can effectively improve the sensitivity and specificity of mass spectrometry detection of fatty acids, and can realize simultaneous quantitative analysis of short-chain, medium-chain, long-chain and ultra-long-chain fatty acids.

Description

technical field [0001] The invention relates to the technical field of analysis and detection. More specifically, the present invention relates to a method for LC-MS / MS analysis of fatty acids based on double derivatization technology. Background technique [0002] Fatty acid (FA) is a class of carboxylic acid compounds with aliphatic chains. According to the length of the aliphatic chain, fatty acids can be divided into: short-chain fatty acids (SCFAs, ≤6 carbon atoms), medium-chain fatty acids (MCFAs, 7-12 carbon atoms), long-chain fatty acids (LCFAs, 13-21 carbon atoms) and Very long chain fatty acids (VLCFAs, ≥ 22 carbon atoms). According to whether the carbon-carbon double bond is divided into saturated fatty acids and unsaturated fatty acids. Fatty acids play a variety of key roles in organisms: (1) Fatty acids are an important part of various functional lipid molecules that constitute biological membranes; (2) Fatty acids can supply energy through β-oxidation, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72G01N2030/067
Inventor 刘丽宏胡婷李晗安卓玲
Owner BEIJING CHAOYANG HOSPITAL CAPITAL MEDICAL UNIV
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