Detection method and application for high-abundance and low-abundance class phospholipid compounds in cells

A technology for phospholipid compounds and detection methods, applied in the fields of biological sample materials and analysis, to achieve the effects of improving the detection rate, expanding phospholipid omics information, and avoiding interference

Pending Publication Date: 2021-06-01
上海鹿明生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These structural diversity brings certain challenges to the analysis and identification of phospholipids

Method used

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  • Detection method and application for high-abundance and low-abundance class phospholipid compounds in cells
  • Detection method and application for high-abundance and low-abundance class phospholipid compounds in cells
  • Detection method and application for high-abundance and low-abundance class phospholipid compounds in cells

Examples

Experimental program
Comparison scheme
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Embodiment

[0078] Detection of high and low abundance phospholipids in RAW264.7 cells

[0079] (1) Pretreatment of RAW264.7 cells

[0080] Take the cells growing in the logarithmic phase, discard the medium, wash the cells twice with PBS, add 1.5 mL of PBS solution pre-cooled at 4°C, blow and beat the cells several times, prepare the cell suspension and count. Take 1 mL of cell suspension in a 1.5 mL centrifuge tube, centrifuge at low temperature, discard the supernatant, and add 40 μL of internal standard solution (PC 17:0-17:0, Lyso PC 17:0, PA 17:0-17:0, LPA17:0, PS 17:0-17:0, LPS 17:1, LPI 17:1, PG 15:0-15:0, PE 17:0-17:0, LPE 17:1) and 1.2mL0. 1N hydrochloric acid aqueous solution / methanol / chloroform mixed solution (1 / 1 / 1, v / v / v), vortex at 2500r for 2 minutes.

[0081] The preparation method of the 0.1N hydrochloric acid aqueous solution is to add 0.9mL concentrated hydrochloric acid into a 100mL volumetric flask, and dilute to the mark with water.

[0082]After standing at 4°C ...

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Abstract

The invention belongs to the technical field of biological sample materials and analysis, and discloses a detection method for high-abundance and low-abundance class phospholipid compounds in cells. According to the method, the UPLC-QqQ MS technology is adopted, and the phospholipid components in the cells are rapidly and effectively separated through UPLC; the glycerophosphatide components in cells are analyzed in a multi-reaction monitoring mode by adopting an ESI positive and negative ion scanning mode, and 218 phospholipid compounds are detected in total, including 44 PC and LPC; 37 kinds of PE and LPE; 38 kinds of PI and LPI; 24 kinds of PG; and 27 kinds of PA and LPA. The method has the characteristics of high specificity and high sensitivity, can be used as an effective means for separation and analysis of complex biological systems such as cells, serum and the like, and lays a foundation for research of phospholipid metabonomics.

Description

technical field [0001] The invention belongs to the field of biological sample materials and analysis technology, and relates to a detection method and application of high-abundance and low-abundance phospholipid compounds in cells, which can quickly and accurately analyze a series of high-abundance and low-abundance phospholipid compounds in cells . Background technique [0002] Phospholipids are the main components of cell membranes and participate in many important metabolic activities such as cell signal transduction, membrane fixation and substrate transport. Many studies have shown that phospholipids play an important role in the occurrence and development of many diseases such as malignant tumors, Alzheimer's disease and obesity, and can be used as potential biomarkers. Phospholipids can be divided into two categories: glycerophospholipids and sphingomyelins, among which glycerophospholipids can be divided into phosphatidylcholine (PC), phosphatidylserine (PS), phosp...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 潘喆敏胡绪俊季佩佩胡哲舒烈波
Owner 上海鹿明生物科技有限公司
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