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An Efficient Single Phase Method for the Extraction of Plasma Lipids.

Alshehry ZH, Barlow CK, Weir JM, Zhou Y, McConville MJ, Meikle PJ - Metabolites (2015)

Bottom Line: Consequently, high throughput methodology that can efficiently extract a wide range of lipids from biological samples is required.Plasma (10 μL) was mixed with 100 μL 1-butanol:methanol (1:1 v/v) containing internal standards resulting in efficient extraction of all major lipid classes (including sterols, glycerolipids, glycerophospholipids and sphingolipids).Lipids were quantified using positive-ion mode LC ESI-MS/MS.

View Article: PubMed Central - PubMed

Affiliation: Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia. malcolmm@unimelb.edu.au.

ABSTRACT
Lipidomic approaches are now widely used to investigate the relationship between lipid metabolism, health and disease. Large-scale lipidomics studies typically aim to quantify hundreds to thousands of lipid molecular species in a large number of samples. Consequently, high throughput methodology that can efficiently extract a wide range of lipids from biological samples is required. Current methods often rely on extraction in chloroform:methanol with or without two phase partitioning or other solvents, which are often incompatible with liquid chromatography electrospray ionization-tandem mass spectrometry (LC ESI-MS/MS). Here, we present a fast, simple extraction method that is suitable for high throughput LC ESI-MS/MS. Plasma (10 μL) was mixed with 100 μL 1-butanol:methanol (1:1 v/v) containing internal standards resulting in efficient extraction of all major lipid classes (including sterols, glycerolipids, glycerophospholipids and sphingolipids). Lipids were quantified using positive-ion mode LC ESI-MS/MS. The method showed high recovery (>90%) and reproducibility (%CV < 20%). It showed a strong correlation of all lipid measures with an established chloroform:methanol extraction method (R2 = 0.976). This method uses non-halogenated solvents, requires no drying or reconstitution steps and is suitable for large-scale LC ESI-MS/MS-based lipidomic analyses in research and clinical laboratories.

No MeSH data available.


Related in: MedlinePlus

Recovery of lipids with the different extraction methods. The recovery calculated the percent of recovery of the method (n = 5) against its spiked equivalent method (n = 5). The black bars represent the recovery of chloroform/methanol against spiked chloroform/methanol, the gray bars represent the recovery of 1-butanol/methanol (3:1 v/v) against spiked 1-butanol/methanol (3:1 v/v) and the white bars represent the recovery of 1-butanol/methanol (1:1 v/v) against spiked 1-butanol/methanol (1:1 v/v).
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metabolites-05-00389-f001: Recovery of lipids with the different extraction methods. The recovery calculated the percent of recovery of the method (n = 5) against its spiked equivalent method (n = 5). The black bars represent the recovery of chloroform/methanol against spiked chloroform/methanol, the gray bars represent the recovery of 1-butanol/methanol (3:1 v/v) against spiked 1-butanol/methanol (3:1 v/v) and the white bars represent the recovery of 1-butanol/methanol (1:1 v/v) against spiked 1-butanol/methanol (1:1 v/v).

Mentions: The recovery of ISTDs from the 1-butanol/methanol (1:1 v/v) were greater than 95% for all ISTDs except LPE (14:0) and CE (18:0) (d6) (90%), and was either similar to, or better than, the recoveries from the chloroform/methanol (2:1 v/v) or 1-butanol/methanol (3:1 v/v) methods (Figure 1). The chloroform/methanol method typically gave higher recovery than the 1-butanol/methanol (3:1 v/v) method except for Cholesterol (d7) (80% and 93%, respectively) and LPE (14:0) (80% and 87%, respectively).


An Efficient Single Phase Method for the Extraction of Plasma Lipids.

Alshehry ZH, Barlow CK, Weir JM, Zhou Y, McConville MJ, Meikle PJ - Metabolites (2015)

Recovery of lipids with the different extraction methods. The recovery calculated the percent of recovery of the method (n = 5) against its spiked equivalent method (n = 5). The black bars represent the recovery of chloroform/methanol against spiked chloroform/methanol, the gray bars represent the recovery of 1-butanol/methanol (3:1 v/v) against spiked 1-butanol/methanol (3:1 v/v) and the white bars represent the recovery of 1-butanol/methanol (1:1 v/v) against spiked 1-butanol/methanol (1:1 v/v).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495379&req=5

metabolites-05-00389-f001: Recovery of lipids with the different extraction methods. The recovery calculated the percent of recovery of the method (n = 5) against its spiked equivalent method (n = 5). The black bars represent the recovery of chloroform/methanol against spiked chloroform/methanol, the gray bars represent the recovery of 1-butanol/methanol (3:1 v/v) against spiked 1-butanol/methanol (3:1 v/v) and the white bars represent the recovery of 1-butanol/methanol (1:1 v/v) against spiked 1-butanol/methanol (1:1 v/v).
Mentions: The recovery of ISTDs from the 1-butanol/methanol (1:1 v/v) were greater than 95% for all ISTDs except LPE (14:0) and CE (18:0) (d6) (90%), and was either similar to, or better than, the recoveries from the chloroform/methanol (2:1 v/v) or 1-butanol/methanol (3:1 v/v) methods (Figure 1). The chloroform/methanol method typically gave higher recovery than the 1-butanol/methanol (3:1 v/v) method except for Cholesterol (d7) (80% and 93%, respectively) and LPE (14:0) (80% and 87%, respectively).

Bottom Line: Consequently, high throughput methodology that can efficiently extract a wide range of lipids from biological samples is required.Plasma (10 μL) was mixed with 100 μL 1-butanol:methanol (1:1 v/v) containing internal standards resulting in efficient extraction of all major lipid classes (including sterols, glycerolipids, glycerophospholipids and sphingolipids).Lipids were quantified using positive-ion mode LC ESI-MS/MS.

View Article: PubMed Central - PubMed

Affiliation: Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia. malcolmm@unimelb.edu.au.

ABSTRACT
Lipidomic approaches are now widely used to investigate the relationship between lipid metabolism, health and disease. Large-scale lipidomics studies typically aim to quantify hundreds to thousands of lipid molecular species in a large number of samples. Consequently, high throughput methodology that can efficiently extract a wide range of lipids from biological samples is required. Current methods often rely on extraction in chloroform:methanol with or without two phase partitioning or other solvents, which are often incompatible with liquid chromatography electrospray ionization-tandem mass spectrometry (LC ESI-MS/MS). Here, we present a fast, simple extraction method that is suitable for high throughput LC ESI-MS/MS. Plasma (10 μL) was mixed with 100 μL 1-butanol:methanol (1:1 v/v) containing internal standards resulting in efficient extraction of all major lipid classes (including sterols, glycerolipids, glycerophospholipids and sphingolipids). Lipids were quantified using positive-ion mode LC ESI-MS/MS. The method showed high recovery (>90%) and reproducibility (%CV < 20%). It showed a strong correlation of all lipid measures with an established chloroform:methanol extraction method (R2 = 0.976). This method uses non-halogenated solvents, requires no drying or reconstitution steps and is suitable for large-scale LC ESI-MS/MS-based lipidomic analyses in research and clinical laboratories.

No MeSH data available.


Related in: MedlinePlus