Electrospray ionization mass spectroscopic analysis of human erythrocyte plasma membrane phospholipids
X Han, R W Gross, X Han, R W Gross
Abstract
Electrospray ionization mass spectrometry (ESI-MS) was utilized for the structural determination and quantitative analysis of individual phospholipid molecular species from subpicomole amounts of human erythrocyte plasma membrane phospholipids. The sensitivity of ESI-MS was 2-3 orders of magnitude greater than that achievable with fast-atom bombardment mass spectrometry (FAB-MS). Phospholipid structure determination and quantitative analysis with ESI-MS can be performed directly from chloroform extracts of biologic samples, obviating the need for prior chromatographic separation of phospholipid classes which has been necessary in FAB-MS phospholipid analyses. Furthermore, ESI-MS is uncomplicated by differential fragmentation of molecular ions and idiosyncratic surface desorption, allowing the quantitation of phospholipids with coefficients of determination (r2) > 0.99 and accuracies > 95%. More than 50 human erythrocyte plasma membrane phospholipid constituents were identified by direct ESI-MS analysis of chloroform extracts of plasma membranes derived from the equivalent of < 1 microliter of whole blood. The major ethanolamine glycerophospholipid subclass in erythrocyte plasma membranes was plasmenylethanolamine that was highly enriched in polyunsaturated fatty acids at the sn-2 position. Collectively, these results demonstrate that ESI-MS of phospholipids is an enabling strategy for the direct structural determination and quantitative analysis of subpicomole amounts of phospholipids from biologic samples.
References
- Methods Enzymol. 1974;31:172-80
- Can J Biochem Physiol. 1959 Aug;37(8):911-7
- Biochemistry. 1984 Jan 3;23(1):158-65
- Biomed Mass Spectrom. 1984 Mar;11(3):132-41
- Circ Res. 1984 Nov;55(5):585-94
- Biochemistry. 1985 Mar 26;24(7):1662-8
- Lipids. 1986 Sep;21(9):580-8
- Biochem Biophys Res Commun. 1987 Jan 15;142(1):194-9
- Nature. 1988 Aug 25;334(6184):661-5
- Proc Natl Acad Sci U S A. 1989 May;86(10):3479-83
- Science. 1989 Oct 6;246(4926):64-71
- Biomed Environ Mass Spectrom. 1990 Mar;19(3):164-70
- Anal Chem. 1990 May 1;62(9):882-99
- Anal Biochem. 1990 Nov 15;191(1):100-5
- FASEB J. 1991 Apr;5(7):2068-77
- FEBS Lett. 1991 Nov 4;292(1-2):201-4
- Prog Lipid Res. 1991;30(4):301-22
- Anal Biochem. 1992 Jan;200(1):119-24
- Rapid Commun Mass Spectrom. 1991 Jul;5(7):309-11
- Anal Chem. 1992 Dec 1;64(23):2965-71
- Biochemistry. 1993 Mar 2;32(8):2031-5
- Anal Chem. 1993 Dec 15;65(24):3669-76
- Science. 1983 May 6;220(4597):568-75
Source: PubMed