Absolute quantitative analysis method of lysophosphatidyl choline based on HPLC-MS/MS detection platform

A technology for quantitative analysis of phosphatidylcholine, applied in the field of medical testing, can solve the problems that LPC substances cannot be popularized, unrealistic, unsatisfactory results of quality control and performance verification, etc.

Active Publication Date: 2017-08-22
TIANJIN CITY THIRD CENT HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is not realistic to draw a corresponding standard curve for all metabolites or find a universal internal standard substance, so a large number of non-targeted metabolism studies and broad-spectrum targeted metabolism studies use the original data Ionic strength is used to represent the level of metabolites, that is, only relative quantification can be ...

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  • Absolute quantitative analysis method of lysophosphatidyl choline based on HPLC-MS/MS detection platform
  • Absolute quantitative analysis method of lysophosphatidyl choline based on HPLC-MS/MS detection platform
  • Absolute quantitative analysis method of lysophosphatidyl choline based on HPLC-MS/MS detection platform

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1 HPLC-MS / MS methodology establishment

[0056] 1. Reagents and instruments

[0057] The sample analysis process mainly involves sample pretreatment, chromatographic separation, and mass spectrometry identification, and the reagents and instruments used are mainly required in these three steps.

[0058] 1.1 Reagents and instruments required for specimen pretreatment

[0059] Reagents: (1) Methanol (manufactured by Honeywell, LC-MS grade in purity), (2) 2,6-di-tert-butyl-p-cresol (2,6-di-tert-butyl-p-cresol also known as ButylatedHydroxytoluene , BHT), the purity of this reagent is superior grade, produced by Bailingwei Enterprise Co., Ltd.

[0060] Instrument: Cleanert PPT 96-well protein precipitation plate, catalog number 96CD2025Q; Cleanert M96 biological sample pretreatment instrument; V96 sample nitrogen blowing concentrator and 96-well collection plate. The above instruments are all from Tianjin Bona Aegel Technology Co., Ltd.

[0061] 1.2 Rea...

Embodiment 2

[0088] Example 2 HPLC-MS / MS Methodology Verification

[0089] Since it is difficult to obtain a matrix completely free of LPC 14:0, LPC 15:0, LPC 16:0, LPC 17:0 and LPC 18:0, the present invention first uses 0.9% normal saline instead of human serum for methodological evaluation and performance verification.

[0090] 1. Verification method

[0091] (1) Evaluation of linear relationship and sensitivity

[0092] Weigh 1.875mg, 3.750mg, 56.250mg, 5.625mg and 37.500mg of LPC 14:0, LPC 15:0, LPC 16:0, LPC 17:0 and LPC 18:0 standards and dissolve them in 50mL methanol solution , take 5 μL and transfer it to 20 μL methanol solution, that is, make the standard curve sample of S7 concentration in Table 3, and use the standard curve sample of S7 concentration to dilute the standard curve sample of each gradient concentration of S1-S6 for standard curve determination and Linear relationship verification. Dilute the standard curve sample of S1 concentration to a lower concentration ...

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Abstract

The invention discloses an absolute quantitative analysis method of lysophosphatidyl choline based on an HPLC-MS/MS detection platform. The absolute quantitative analysis method establishes the HPLC-MS/MS detection platform, takes reserpine as an internal standard to draw the standard curves of LPC 14:0-LPC 18:0 serial substances, verifies the linearity, precision and accuracy of the detection platform, and detects the matrix effect of blood serum. Proven by verification, the HPLC-MS/MS detection platform established by the invention has higher stability and accuracy in detection of the LPC substances in the human blood serum, meets the clinical requirements and is suitable for clinical popularization.

Description

technical field [0001] The invention belongs to the field of medical testing, and relates to an absolute quantitative analysis method for lysophosphatidylcholine, in particular to an absolute quantitative analysis method for lysophosphatidylcholine based on an HPLC-MS / MS detection platform. Background technique [0002] HBV-related HCC is the most common type of liver cancer in my country, and the occurrence and development of HCC is a complex pathological process involving abnormal changes in multiple signaling pathways and metabolic pathways. During this process, the metabolic levels of various substances change. Metabolites have clinical value for potential diagnosis, disease staging, and prognosis. Among them, LPC substances have been reported many times to have a certain relationship with HCC. [0003] Lysophosphatidylcholine (LPC) is the main component of oxidized low density lipoprotein (oxidized low density lipopro2 tei ox2LDL), one of the main products of phospholip...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 刘树业张磊范志娟康华王宇凡田亚琼刘爽武玉晶
Owner TIANJIN CITY THIRD CENT HOSPITAL
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