Recent Analytical Methodologies in Lipid Analysis
Abstract
:1. Introduction
2. Sample Pretreatment
2.1. Extraction of Lipids
2.2. Derivatization
3. Instrumental Analysis of Lipids
3.1. Direct Infusion MS
3.2. Mass Spectrometry Imaging
3.3. Ion Mobility Spectrometry (IMS-MS)
3.4. LC-MS
3.5. Supercritical Fluid Chromatography—Mass Spectrometry (SFC-MS)
3.6. Gas Chromatography—Mass Spectrometry (GC-MS)
Sample | Analytes | Extraction Type | Extraction Solvent | Method | Approach | Results | Ref. |
---|---|---|---|---|---|---|---|
Biological samples | |||||||
rat serum, brain tissue | SP | LLE | CHCl3:MeOH (9:1) | RPLC-MS/MS | targeted | method for quantification of SP in biological samples | [138] |
human plasma, mouse serum | lipidomic profiling | BUME | BuOH:MeOH (1:1) | LC-MS/MS | untargeted | 88 lipid species were identified as significantly different between wild type CerS2 null mice | [139] |
human serum | lipid profiling | LLE | CHCl3:MeOH (3:1) | UHPLC-HRMS | untargeted | potentially 12 lipids can serve as diagnostic markers of colorectal adenoma | [140] |
serum | HDL | LLE (Folch method) | CHCl3:MeOH | LC-MS/MS | targeted | association of MetS with impairment of phospholipid metabolism in HDL, with obesity and insulin resistance | [141] |
plasma | SP | OPE | MeOH | LC-MS/MS | targeted | 33 identified SP | [142] |
mouse tissue | lipid profiling | OPE | MeOH:H2O (80:20) | LC-MS/MS | - | identification of major cardiolipin molecular species by BRI-DIA and hybrid methods | [143] |
rat serum | lipid markers of CHD | LLE | MTBE | UPLC-HDMS | - | GP and SP metabolism as targets for the treatment of CHD | [144] |
porcine brain extract | lipidomic profile | LLE | MTBE | RP-LC-MS | - | development of microgradient fractionation of total lipid extract for lipidomic analysis. | [145] |
renal biopsies | lipid biomarkers of Fabry disease | LLE (Folch method) | CHCl3:MeOH | UHPLC-HRMS | untargeted | identification of biomarkers of Fabry disease | [146] |
pancreatic cancer cells, extracellular vesicles | lipids and metabolites | LLE | CHCl3:MeOH | SFC-MS | - | identification of 494 lipids | [135] |
human serum | PCs | SPE | eluted with IPA | LC-MS/MS | - | elevation of oxidized PCs in the acute phase of KD | [147] |
human cancer cells and EVs | lipidomic profile | LLE (Bligh and Dyer method) | CHCl3:MeOH | SFC-MS | - | breast cancer EVs selectively loaded with lipids supporting tumor progression | [148] |
human plasma | polar lipids | OPE | MeOH | LC-MS/MS | method development for monitoring of 398 polar lipids | [149] | |
plasma, urine | oxidation products of PUFA | LLE (Folch method) | CHCl3:MeOH | LC-QTOF-MS/MS | targeted | method development for measuring of oxidation products of PUFA | [150] |
human CSF | VLCFA | SPE, LLE + derivatization | octane:EtOH (88:12) + DAABD-AE | UPLC-MS/MS | targeted | assay development for measuring of VLCFA biomarkers | [151] |
human plasma | lipidome | LLE, UAE | CHCl3:MeOH (3:1) | UHPLC-MS | targeted, untargeted | PC (18:1/P-16:0), PC (o-22:3/22:3), PC(P-18:1/16:1) as biomarkers of metabolic syndrome | [152] |
human plasma | lipidomic biomarkers | OPE | IPA | LC-MS | targeted | reference for bladder cancer and renal cell carcinoma biomarker discovery | [153] |
human fibroblasts | unsaturated FA | LLE | MTBE | LC-MS | targeted | complete characterization of FA species | [154] |
mouse plasma | CE, FA, PC, NAE, SM | LLE (Folch method) | CHCl3:MeOH | UHPLC-HR-MS | untargeted | identification of plasma lipid species associated with pain and/or pathology in a DMM model of OA | [155] |
human plasma | LPCs | OPE, UAE | MeOH:ACN | LC-ESI-MS/MS | targeted | identification of 60 LPCs | [156] |
human plasma | lipidomic screening | LLE (Bligh and Dyer method) | CH3OH–CH2Cl2 | UPLC-MS | untargeted | increasing of TAGs levels of advanced-stage CRC patients compared with early-stage CRC patients | [157] |
human serum | LPC, PC, LPE, PE, LPS, PS, LPG, PG, LPI, PI, LPA, PA, SM, MAG, DAG, TAG, CL, Cer, CE | LLE (Folch method) | CHCl3:MeOH (2:1, v/v) | RPLC-MS/MS | untargeted | identification of 753 lipids | [158] |
mouse tissues and fluids | acylcarnitines | OPE + derivatization | MeOH:H2O + 3-NPH | LC-MS | targeted | identification of 123 acylcarnitines | [159] |
plasma, fecal | SCFAs | OPE + derivatization | H2O + 2- bromoacetophenone | LC-MS/MS | targeted | identification of 7 SCFAs | [160] |
plasma, tissue | lipid mediators | SPE | eluted with methyl formate | LC-MS/MS | untargeted | novel tool for studying complete profile of lipid mediators in biological samples | [161] |
human serum | lysosphingomyelin-509 | OPE | EtOH:H2O (3:1, v/v) | LC-MS | targeted | identification of lysosphingomyelin-509 | [162] |
mouse liver | lipid profile | LLE | MeOH:DCM (1:3) | UPLC-MS | - | significant differences in lipid profiles of SCID and chimeric PXB liver-humanized mice | [163] |
Food | |||||||
green, red lettuce | sulfolipids, galactolipids | LLE (Folch method) | CHCl3:MeOH (3:2) | LC-ESI-MS/MS | targeted | oxidized SQDG as potential markers for abiotic stress factors | [164] |
geopropolis | lipid profiles | LLE | MeOH, CHCl3 | LC-HRMS | - | identification of 61 lipids | [165] |
oil palm | lipid profiles | LLE | MTBE | LC-MS | targeted | lipidomic tools for analysis of lipid composition variability in oil from palm | [166] |
fish oil, mushroom extract | FuFA-containing TAGs | LLE, UAE | cyclohexane:EtOAc (46:54)IPA:n-hexane (1:4) | LC-HRMS | - | identification of 39 different FuFA-containing TAGs | [167] |
olive fruit seeds | polar lipids | LLE (Folch method) | CHCl3:MeOH (2:1) | HILIC-HR-MS/MS | untargeted | identification of 94 lipids | [168] |
coffee | specific lipids of interest for each coffee origin | LLE | MTBE | LC-MS/MS | targeted | determination of coffee origin based on its lipid profile | [169] |
donkey meat | lipid profiles | LLE (Folch method) | CHCl3:MeOH (2:1) | LC-MS | untargeted | identification of 1143 lipids | [170] |
milk | HFAs | OPE | MeOH | LC-HRMS | - | quantification of 19 free HFAs | [171] |
extra virgin olive oil | FFAs, FFA methyl- and ethylesters, MAGs, triterpenoids, TAGs | OPE | IPA | LC-MS/MS | - | potent tool for studying variability of lipid species in olive oil | [172] |
potatoes | polar lipids | LLE (Bligh and Dyer, Folch, ”Green” Folch, Matyash, extraction with n-hexane) | CHCl3:MeOH EtOAc:MeOH MTBE n-hexane | UPLC-MS | targeted, untargeted | “Green” Folch method (with EtOAc)—the most suitable extraction method | [173] |
Pharmaceuticals | |||||||
dietary supplements | lipid profiling | - | - | LC-MS | - | production of different lipid classes by different based ingredients products | [174] |
Bacteria | |||||||
Pseudomonas aeruginosa | phospholipids | LLE (Bligh and Dyer) | CHCl3:MeOH | LC-MS/MS | - | the growth medium can influence membrane lipid composition | [175] |
C. eiseniae, Olivibacter sp. | glycerophosholipids | LLE | MTBE MeOH | UHPLC-HR-MS | - | identification of 2 novel glycerophospholipids, 2 novel LAAs | [176] |
Escherichia coli | GPs | LLE | MTBE | UPLC-MS/MS | targeted | transferability of method to any UPLC-MS/MS system with no hardware modification need | [177] |
Fungi | |||||||
marine fungi | ergosterol | LLE (Bligh and Dyer) | CHCl3:MeOH | LC-MS/MS | targeted | highly sensitive method for measuring fungal biomass | [178] |
Plants | |||||||
plant tissue | polar and non-polar lipids | LLE | different solvents optimization of extraction | UHPLC-MS/MS | - | method development for evaluating of polar and non-polar lipids | [125] |
tobacco hairy roots | GPL | LLE (Bligh and Dyer) | CHF3:MeOH | HILIC-MS/MS | targeted | method development for simultaneous determination of different phospholipids | [179] |
Arabidopsis thaliana | lipid profiling | LLE | CHCl3:MeOH:H2O (1:2.5:1) MeOH:MTBE (1:3) IPA + CHCl3:MeOH:H2O (30:41.5:3.5) IPA + CHCl3:H2O (5:2) + CHCl3:MeOH (2:1) | LC-MS | targeted, untargeted | single-step extraction method for untargeted lipidomic analysis | [34] |
4. Data Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Lipid Class | Example |
---|---|
Fatty acyls | HFAs, FAs |
Glycerolipids | MAG, DAG, TAG |
Glycerophospholipids | PA, PC, PE, PS |
Sphingolipids | Cer, PSL, SM |
Sterol lipids | CE, Chol, CS |
Prenol lipids | quinine, polyprenol, isoprenoid |
Saccharolipids | lipid A |
Polyketides | lovastatin |
Extraction Method | Advantages | Disadvantages | Ref. |
---|---|---|---|
OPE | easy to perform possibility of automation low cost precipitation of proteins and insoluble organic species | may not remove interferences efficiently long centrifugation needed | [26,54] |
LLE | well-established protocols many combinations of solvents could be used (in different ratios) low cost | time-consuming difficult to automate repeated extractions needed challenging organic phase layer transfer | [54] |
SPE | reduction of matrix effect purification of samples wide range of commercially available SPE sorbents | particularly suitable for targeted analysis long optimization of washing and elution solvents | [27] |
SPME | requires very small amount of sample reduction of matrix effect small amounts of solvents needed fast extraction | suitable mainly for GC lower extraction ability | [7,27,55] |
UAE | highly-reproducible time-efficient improve the extraction efficiency in combination with LLE | use of toxic solvents longer extraction times increasing the temperature (because of fictions) which leads to degradation possibly damage hearing | [21] |
MAE | improvement of extraction efficiency reduction of time and organic solvents consumption | potential degradation of thermally instable lipids long optimization of extraction parameters | [21] |
SE | provides a high yield of lipids possibility of use with green solvents | continuous heating at the boiling temperature could lead to lipid oxidation and degradation of heat liable, time consuming | [11] |
SFE(SCO2) | shorter extraction times suitable for neutral, low-polarity lipids supercritical CO2 is green solvent | extraction of polar lipids requires use of organic modifier high initial costs of equipment | [11,26,42] |
Sample | Analytes | Extraction Type | Extraction Solvent | Results | Ref. |
---|---|---|---|---|---|
rat brain tissue | lipid profile | OPE | MeOH | direct infusion probe development for metabolomics | [72] |
20 mammalian cells | 19 lipid subclasses | LLE | CHCl3:MeOH:IPA, (1:2:4) | determination of different lipid species with potential for clinical applications | [73] |
fermented vegetable juices | lipid profiling | LLE | MTBE | fermented juices contain more beneficial metabolites and carotenoids than commercial non-fermented juices | [74] |
mammalian samples | lipidome | LLE (Bligh and Dyer) | CHCl3:MeOH | guideline for setting up and using platform for exploring mammalian lipidome | [69] |
bovine milk | TAG | LLE | CHCl3 | identification of more than 100 TAGs | [75] |
MALDI-MSI | ||||
---|---|---|---|---|
Sample | Analytes | Matrix | Results | Ref. |
human kidney tissues | lipidome | DAN | comparing of LIMS and HPLC-MS—identification of larger number of species with using HPLC-MS | [86] |
rat brain tissue | lipidomic profiles | norharmane | lipidomic spectra showed high consistency between MALDI and WALDI | [76] |
human and murine tissue | lipid profiling | DHB | identification of several atherosclerosis- specific lipid biomarkers | [92] |
salivary gland tumor tissue | lipidomic profile | DHB | MALDI-MSI complementary diagnostic tool | [77] |
human tissue | lipid profiling | DHB | plaque features and specific lipid classes clear colocalization | [93] |
osteoarthritic synovial membrane | lipidomic profile | norharmane | novel insight into lipid profiling of synovial membrane | [94] |
colorectal cancer tissue | lipidomic profile | 9-AA | tool for subtyping the diverse immune environments in CRC | [95] |
tumor spheroids | lipid metabolites | DHB | method for detailed information about spheroids and drug relationship | [96] |
human tissue | lipid profiling | DHB | diacylglycerols are more abundant in thrombotic area in comparison with other plaque areas | [97] |
human kidney tissue | Lipid storage | DHB | sections stored at RT (one week of storage)—largest amount of lipid degradation in comparison with sections stored under N2 at −80 °C | [98] |
SIMS-MSI | ||||
Sample | Analytes | Primary Ion Beam | Results | Ref. |
mammalian CMs | lipid profiling | Ar2000+ Bi3+ | identifying of heart failure associated lipids | [83] |
lipid extracts, cells, mouse brain tissue | lipid profiling | (CO2)n+ (H2O)n+, (H2O)n +(CO2) | imaging of LPC for the first time using TOF-SIMS | [99] |
Gammarus fossarum | lipidome characterization | Bi3+ cluster ions | compositional and spatial information of lipids | [100] |
infarcted mouse heart tissue | spatial distribution of lipids | gas cluster ion beam (Ar4000+) | different spatial lipids distributions; insights changes in lipid metabolism following infarction | [101] |
DESI-MSI | ||||
Sample | Analytes | Solvent System, Technique | Results | Ref. |
mice liver tissue | lipid distribution | MeOH:H2O (98:2) DESI | zone-specific hepatic lipid distribution of three zones | [102] |
human carotid plaque | lipid signatures | MeOH:H2O (98:2) DESI | identified lipid species present in plaque (compared with plasma) | [103] |
asiatic toad | lipid composition | MeOH:H2O (95:5) DESI | significant lipid metabolism changes due to body remodeling during metamorphosis | [104] |
xenograft glioblastoma tumour | lipid profiling | MeOH:H2O (95:5) 3D DESI | heterogeneous lipid expression is important to aid β-oxidation in hypoxic areas glioblastoma | [105] |
cow, sow, mouse ovaries | lipid distribution | DMF:ACN (1:1) DESI | similar lipid signatures of corpora lutea, follicular wall, ovarian stroma independent of the species | [106] |
swine fetuses | lipid distribution | DMF:ACN (1:1) DESI | organ-dependent localization of lipids, indication of key lipids related to physiological organogenesis | [107] |
mouse lung tissues | lipid coverage | MeOH:H2O (9:1) nanoDESI | spatial localization of lipids in tissues. 50% of lipid coverage in comparison with Folch extraction-LC-MS/MS method | [91] |
Sample | Analytes | Extraction Type | Extraction Solvent | Method | Results | Ref. |
---|---|---|---|---|---|---|
porcine oocyte | lipidomic profile | LLE | MeOH:CHCl3 | nanoLC-TIMS-MS | oocyte lipids identification and relative quantification at the single-cell level | [120] |
human plasma, serum | lipid profiling | LLE | MTBE:MeOH (10:3) | UHPLC-TIMS-PASEF-MS | Annotation of 370 lipids in reference plasma and 364 lipids in serum sample | [121] |
mouse brain tissue | lipid localization | - | MeOH:H2O (9:1) nanoDESI | nanoDESI-TIMS-MSI | separation of lipid isomers and isobars and localization in brain tissue | [122] |
plasma | lipid profile | LLE | MTBE | UHPLC-TIMS-MS | approach development for untargeted lipidomics | [123] |
human plasma, mouse liver, HeLa cells | lipidomic profile | LLE | MeOH:MTBE:H2O | nanoLC-TIMS-PASEF-MS | 1108 lipids (0.05 μL plasma), 976 lipids (10 μg liver tissue) and 1351 lipids (~2000 HeLa cells) were identified | [124] |
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Gerhardtova, I.; Jankech, T.; Majerova, P.; Piestansky, J.; Olesova, D.; Kovac, A.; Jampilek, J. Recent Analytical Methodologies in Lipid Analysis. Int. J. Mol. Sci. 2024, 25, 2249. https://doi.org/10.3390/ijms25042249
Gerhardtova I, Jankech T, Majerova P, Piestansky J, Olesova D, Kovac A, Jampilek J. Recent Analytical Methodologies in Lipid Analysis. International Journal of Molecular Sciences. 2024; 25(4):2249. https://doi.org/10.3390/ijms25042249
Chicago/Turabian StyleGerhardtova, Ivana, Timotej Jankech, Petra Majerova, Juraj Piestansky, Dominika Olesova, Andrej Kovac, and Josef Jampilek. 2024. "Recent Analytical Methodologies in Lipid Analysis" International Journal of Molecular Sciences 25, no. 4: 2249. https://doi.org/10.3390/ijms25042249