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Method for determining contents of L-carnitine and acetyl-L-carnitine in serum of depression patient by high performance liquid chromatography tandem mass spectrometry

A high-performance liquid chromatography, acetyl-L-carnitine technology, applied in the biological field, can solve time-consuming and labor-intensive problems

Inactive Publication Date: 2021-02-12
合肥市第四人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods require the addition of ion-pairing reagents or solid-phase extraction, which is time-consuming and laborious for sample preparation

Method used

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  • Method for determining contents of L-carnitine and acetyl-L-carnitine in serum of depression patient by high performance liquid chromatography tandem mass spectrometry
  • Method for determining contents of L-carnitine and acetyl-L-carnitine in serum of depression patient by high performance liquid chromatography tandem mass spectrometry
  • Method for determining contents of L-carnitine and acetyl-L-carnitine in serum of depression patient by high performance liquid chromatography tandem mass spectrometry

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Embodiment Construction

[0021] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0022] 1. Instrument and chromatographic conditions

[0023] The UPLC-MS / MS instrument consists of Waters Acquity ultra-high performance liquid chromatography (IPLC) Class I binary solvent manager, Acquity UPLC sample manager-FTN and Waters Xevo TQ-S mass spectrometer. The chromatographic column is an XSelect HSS T3C18 (2.1mm×100 mm, 2.5μm, Waters, USA) analytical column, and the mobile phase is water (mobile phase A, containing 0.01% ammonia water) and methanol (mobile phase B), and the flow rate is 0.2 ml / min, the internal standard is acetyl-L-carnitine-d 3 (ALC-d 3 ), gradient elution, the gradient curve is as follows: 0~0.5min, 20%~20% B; 0.5~1.5min, 20%~95% B; 1.5~2.0min, 95%~95% B; 2.0~2.1min , 95%~20% B, 2.1~2.5min, 20%~20% B, the injection volume is 0.2 μL.

[0024] 2. Mass Spectrometry Conditions

[0025] Ion source: electrospray ion so...

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Abstract

The invention discloses a method for determining the contents of L-carnitine and acetyl-L-carnitine in serum of a depression patient by high performance liquid chromatography tandem mass spectrometry.The method comprises the following steps: S1, drawing a standard curve: preparing ALC with 8 concentrations and LC with 8 concentrations in human serum, constructing a standard curve by adopting baseline subtraction under certain liquid chromatography and mass spectrometry conditions, performing baseline subtraction by operating a blank plasma sample only added with an internal standard (IS) in each batch of measurement, drawing a series of standard curves according to the concentration of an analyte, subtracting the area ratio of the analyte to the IS peak in the blank sample from the area ratio of the analyte to the IS peak of the sample to obtain a vertical coordinate, and linearly fitting a standard curve in a certain concentration range by taking the concentration as a horizontal coordinate; and S2, performing sample processing and computer testing. The content of ALC and LC in serum of a depression patient can be measured at the same time, the steps are simple, and the detectionresult is accurate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for determining the content of L-carnitine and acetyl-L-carnitine in serum of patients with depression by high performance liquid chromatography tandem mass spectrometry. Background technique [0002] L-carnitine (LC) is a key substance in the process of fat metabolism, which can promote the oxidation and decomposition of fatty acids in mitochondria. Acetyl-L-carnitine (ALC) is the natural form of L-carnitine. On the inner mitochondrial membrane, ALC is catalyzed by carnitine acetyltransferase to synthesize acetyl coenzyme A (CoA) and L-carnitine, and enters the mitochondrial matrix through carnitine / acetylcarnitine acyl translocase, while a molecule of carnitine passes through the mitochondria matrix. Carnitine acetyltransferase in the mitochondrial matrix then converts acylcarnitines to carnitine and acetyl-CoA. Acetyl CoA and choline are catalyzed by choline acetyltrans...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 夏清荣梁俊聂丽娟单锋徐亚运闫春宇周旋程琢玉
Owner 合肥市第四人民医院