+ Site Statistics
References:
54,258,434
Abstracts:
29,560,870
PMIDs:
28,072,757
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Comprehensive approach to the quantitative analysis of mitochondrial phospholipids by HPLC-MS



Comprehensive approach to the quantitative analysis of mitochondrial phospholipids by HPLC-MS



Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences 912: 105-114



A normal-phase HPLC-MS method was established to analyze mitochondrial phospholipids quantitatively as well as qualitatively. An efficient extraction procedure and chromatographic conditions were developed using twelve standardized phospholipids and lysophospholipids. The chromatographic conditions provided physical separation of phospholipids by class, and efficient ionization allowed detection of low abundance phospholipids such as phosphatidylglycerol and monolysocardiolipin. The chromatographic separation of each class of phospholipid permitted qualitative identification of molecular species without interference from other classes. This is advantageous for mitochondrial lipidomics because the composition of mitochondrial phospholipids varies depending on tissue source, pathological condition, and nutrition. Using the method, seven classes of phospholipids (phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, cardiolipin, and monolysocardiolipin) were detected in rat heart and skeletal muscle mitochondria and all but phosphatidylserine were quantified. The concentration was calculated using standard curves with an internal standard generated for each class of phospholipid. The method was validated for intraday and interday variation and showed excellent reproducibility and accuracy. This new method, with each step documented, provides a powerful tool for accurate quantitation of phospholipids, a basic structural component of mitochondrial membranes.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 052269996

Download citation: RISBibTeXText

PMID: 23266842

DOI: 10.1016/j.jchromb.2012.10.036


Related references

HPLC analysis of mitochondrial membrane phospholipids in rice. Journal of Analytical Chemistry 62(4): 369-372, 2007

Quantitative analysis of phospholipids by hplc with a light scattering evaporating detector application to raw materials for cosmetic use. HRC Journal of High Resolution Chromatography 13(2): 126-136, 1990

Comprehensive molecular diagnosis of mitochondrial disorders: qualitative and quantitative approach. Annals of the New York Academy of Sciences 1011: 246-258, 2004

Quantitative analysis of phospholipids from whey protein concentrates by HPLC with a narrow-bore column and a light-scattering detector. Journal of Dairy Science 77(SUPPL 1): 12, 1994

Quantitative mitochondrial DNA mutation analysis by denaturing HPLC. Clinical Chemistry 53(6): 1046-1052, 2007

Quantitative analysis combined with chromatographic fingerprint for comprehensive evaluation of Danhong injection using HPLC-DAD. Journal of Pharmaceutical and Biomedical Analysis 76: 70-74, 2013

Comprehensive quantitative analysis of Shuang-Huang-Lian oral liquid using UHPLC-Q-TOF-MS and HPLC-ELSD. Journal of Pharmaceutical and Biomedical Analysis 102: 1-8, 2015

Quantitative analysis combined with chromatographic fingerprint for comprehensive evaluation of Xiaoer Chaigui Tuire granules by HPLC-DAD. Journal of Chromatographic Science 53(5): 749-756, 2015

A versatile LC-MS/MS approach for comprehensive, quantitative analysis of central metabolic pathways. Wellcome Open Research 3: 122, 2018

Studies on the phospholipids in cow's and human milk. 2. Quantitative analysis of phospholipids by thin layer chromatography. J. Jap. Soc. Food Nutrition. 18: 387-390, 1966

In situ sample processing approach (iSPA) for comprehensive quantitative phosphoproteome analysis. Journal of Proteome Research 13(9): 3896-3904, 2015

Comprehensive analysis of phospholipids in the brain, heart, kidney, and liver: brain phospholipids are least enriched with polyunsaturated fatty acids. Molecular and Cellular Biochemistry 442(1-2): 187-201, 2017

Thin layer chromatography of retinal phospholipids. Quantitative and qualitative analysis of phospholipids from normal rabbit's retina. Minerva Oftalmologica 9(1): 18-19, 1967

Purification and analysis of phospholipids in the inner mitochondrial membrane fraction of bovine corpus luteum, and properties of cytochrome P-450scc incorporated into vesicles prepared from these phospholipids. European Journal of Biochemistry 148(2): 379-384, 1985

Phospholipids in tissues of the eye. I. Isolation, characterization and quantitative analysis by two-dimensional thin-layer chromatography of diacyl and vinyl-ether phospholipids. Biochimica et Biophysica Acta 152(2): 307-315, 1968