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Absolute Quantitative Analysis Method of Lysophosphatidylcholine Based on HPLC-MS/MS Detection Platform

A phosphatidylcholine, quantitative analysis technology, applied in the field of medical testing, can solve the problems of unsatisfactory quality control and performance verification results, unrealistic, and inability to popularize and apply LPC substances

Active Publication Date: 2019-03-15
TIANJIN CITY THIRD CENT HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 performed
At present, some studies have tried to use simplified standardized methods (such as common internal standards) to solve quantitative problems, but the results of their quality control and performance verification are not satisfactory
Due to the defects of LC-MS technology itself, it cannot be widely applied to the detection of LPC substances in clinical practice.

Method used

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  • Absolute Quantitative Analysis Method of Lysophosphatidylcholine Based on HPLC-MS/MS Detection Platform
  • Absolute Quantitative Analysis Method of Lysophosphatidylcholine Based on HPLC-MS/MS Detection Platform
  • Absolute Quantitative Analysis Method of Lysophosphatidylcholine Based on HPLC-MS/MS Detection Platform

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Experimental program
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Effect test

Embodiment 1

[0055] Example 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. The reagents and instruments used are also mainly required in these three steps.

[0058] 1.1 Reagents and instruments required for specimen pretreatment

[0059] Reagents: (1) methanol (manufactured by Honeywell, with a purity of LC-MS grade), (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 excellent grade pure, produced by Bailingwei Enterprise Co., Ltd.

[0060] Equipment: Cleanert PPT 96-well protein precipitation plate, item number 96CD2025Q; Cleanert M96 biological sample pre-processing instrument; V96 sample nitrogen blowing concentrator and 96-well collection plate. The above instruments are from Tianjin Bona Aijieer Technology Co., Ltd.

[0061] 1.2 Reagents a...

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 LPC 14:0, LPC 15:0, LPC 16:0, LPC 17:0 and LPC 18:0 standard products 1.875mg, 3.750mg, 56.250mg, 5.625mg and 37.500mg and dissolve in 50mL methanol solution , Take 5μL and move it to 20μL methanol solution to prepare the standard curve sample of S7 concentration in Table 3. Use the standard curve sample of S7 concentration to dilute to 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 the S1 concentration to a lower concentration gradient for comp...

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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 detection, and relates to an absolute quantitative analysis method of lysophosphatidylcholine, in particular to an absolute quantitative analysis method of 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 multiple substances change. Metabolites have clinical value for potential diagnosis, disease staging and prognosis. Among them, LPC substances have been repeatedly reported to have a certain relationship with HCC. [0003] Lysophosphatidylcholine (LPC) is the main component of oxidized low density lipopro2 tei ox2LDL, and one of the main products of phospholipid catabolism, and its content in huma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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