Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Large-scale blood plasma sphingolipid profile analysis method based on liquid chromatography-mass spectrometry combination

A liquid chromatography and profile analysis technology, applied in the field of large-scale profile analysis of plasma sphingolipids, can solve problems such as complex methods, achieve the effects of simple and rapid operation, low solvent toxicity, and improved detection sensitivity

Active Publication Date: 2014-06-25
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF1 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the diversity in structure and abundance of each subclass of sphingolipids, the introduction of multiple platforms (multiple extractions + multiple LC-MS methods) can cover as many sphingolipids as possible, but will complicate the method

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Large-scale blood plasma sphingolipid profile analysis method based on liquid chromatography-mass spectrometry combination
  • Large-scale blood plasma sphingolipid profile analysis method based on liquid chromatography-mass spectrometry combination
  • Large-scale blood plasma sphingolipid profile analysis method based on liquid chromatography-mass spectrometry combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Thaw normal human plasma stored at -80°C in an ice bath, pipette 40 μL quantitatively, add 30 μL internal standard mixed solution, and vortex to mix. Internal standard solutions include Ceramide(d18:1 / 17:0) 0.42nmol / mL, SM(d18:1 / 12:0) 1.67nmol / mL, Sphingosine(d17:1) 0.17nmol / mL, Lactosylceramide(d18:1 / 12:0) 0.17nmol / mL, Glucosylceramide (d18:1 / 12:0) 0.03nmol / mL, Dihydroceramide (d18:0 / 12:0) 0.03nmol / mL. Add 225 μL of methanol and 75 μL of 1M sodium methoxide-methanol solution and vortex for 30 s. Add 1mL MTBE, vortex 30s. Shake on a shaker at 37°C for 2h. After taking it out and cooling it, add 25 μL of 20% (v / v) acetic acid-water solution and 250 μL of ultrapure water. Vortex for 30s. Centrifuge at 10°C at 12,000 rpm for 10 min, remove 800 μL of the supernatant quantitatively, and freeze-dry. With complex solution methanol / isopropanol / water=65:30:5 (MeOH / IPA / H 2 O=65:30:5) reconstituted to 50 μL, vortexed for 1 min, and used for LC-MS / MS analysis, the detection ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a large-scale blood plasma sphingolipid profile analysis method based on liquid chromatography-mass spectrometry combination, which is characterized in that sphingolipid in blood plasma is extracted by double-phase extraction with methyl tertiary butyl ether / methanol / water (MTBE / MeOH / H2O) combined with mild basic hydrolysis. Then rapid semi-quantitative analysis is realized within 15 min by combination of ultra-high performance liquid chromatography-electro-spray ionization-mass spectrometry. In the invention, the MTBE / MeOH / H2O extraction system is simple, rapid, and easy to operate, has less protein interference in the extract, and has good repeatability; interference of high-abundance phospholipid and glyceride in the lipid group is eliminated by hydrolysis; ion inhibition is reduced; therefore, the purpose of detecting low-abundance sphingolipid with high sensitivity is achieved. With the high resolution separation capability of subsequent ultra-high performance liquid chromatography (UHPLC) and the specificity of MRM, effective distinguishing of isomers is realized, and the introduction of complex isotope correction during quantification is avoided, which makes the method more rapid and accurate.

Description

technical field [0001] The invention relates to the fields of analytical chemistry and medicine, and is a large-scale plasma sphingolipid profile analysis method based on liquid chromatography-mass spectrometry. Background technique [0002] Sphingolipids (SLs) are a class of lipid molecules with 1,3-hydroxy-2-aminoalkanes / alkenes as the common core. As the 2-amino group is connected to various fatty acids with different chain lengths and saturations through amide bonds, and the 1-hydroxy group is connected to various polar groups (such as phosphorylcholine, various simple and complex sugar groups, etc.), The molecular structure of sphingolipids is complex and diverse, and the chemical properties are different. At the same time, the abundance of each subclass of sphingolipids also varies greatly. Although the sphingolipid group is a small part of the whole lipid group, its function in living organisms cannot be underestimated. Sphingolipids are an essential component of th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N30/02
Inventor 许国旺李佳赵欣捷路鑫
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com