Thin-layer chromatographic analysis of lipids and lipophilic pigments in snails

Fried, Bernard; Sherma, Joseph

doi:10.1556/1006.2016.29.2.2

Cited by 4 publications

(3 citation statements)

References 45 publications

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“…To resolve and quantify fast moving neutral lipid bands (squalene, ME, and SE), plates were developed with the Smith mobile phase: hexane-petroleum ether-diethyl ether-glacial acetic acid (50:20:5:1, v / v / v / v ). The use of these mobile phases in HPTLC was recently reviewed in Bui et al [ 24 ]. One-dimensional ascending development was carried out in an HPTLC twin trough chamber (CAMAG) containing a saturation pad (Miles Scientific).…”

Section: Methodsmentioning

confidence: 99%

“…In a previous study done by Schmidt et al [ 22 ], the lipid content of lipid droplets in yeast was analyzed using only qualitative one-dimensional thin-layer chromatography (TLC) with double development for nonpolar lipids and qualitative two-dimensional TLC for phospholipids, showing the major lipid classes in yeast to be free sterol (FS), free fatty acid (FFA), triacylglycerol (TAG), and steryl ester (SE) and phosphatidylinositol (PI), phosphatidylethanolamine (PE), and phosphatidylcholine (PC), respectively. HPTLC (high performance TLC) has been utilized extensively in lipid quantification in biological samples (reviewed by Fried [ 23 ] and Bui et al [ 24 ]). Recently, we showed that HPTLC was a robust method of quantifying lipids in yeast cells, allowing highly reproducible and accurate measurements [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

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Using High Performance Thin Layer Chromatography-Densitometry to Study the Influence of the Prion [RNQ+] and Its Determinant Prion Protein Rnq1 on Yeast Lipid Profiles

Sherma

Hines

2018

Separations

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The baker’s yeast Saccharomyces cerevisiae harbors multiple prions that allow for the creation of heterogeneity within otherwise clonal cell populations. However, in many cases, the consequences of prion infection are entirely unclear. Predictions of prion-induced changes in cell physiology are complicated by pleotropic effects, and detection is often limited to relatively insensitive cell growth assays that may obscure many physiological changes. We previously showed that silica gel high performance thin-layer chromatography-densitometry (HPTLC) can be used to empirically determine prion-induced changes in lipid content in yeast. Here, we conduct pair-wise quantifications of the relative levels of free sterols, free fatty acids, and triacylglycerols [petroleum ether-diethyl ether-glacial acetic acid (80:20:1, v/v/v) mobile phase and phosphomolybdic acid (PMA) detection reagent]; steryl esters, methyl esters, and squalene [hexane-petroleum ether-diethyl ether-glacial acetic acid (50:20:5:1, v/v/v/v) and PMA]; and phosphatidylethanolamine, phosphatidylcholine, and phosphatidylinositol (chloroform-diethyl ether-acetic acid (65:25:4.5, v/v/v) and cupric sulfate-phosphoric acid) in otherwise clonal prion-infected ([RNQ+]) and prion-free ([rnq−]) cells in both stationary- and logarithmic-growth phases. We detected multiple statistically significant differences between prion-infected and prion-free cells that varied by growth phase, confirming our pr evious observations that prions exert distinct influences on cell physiology between stationary- and log-phase growth. We also found significant differences between cells expressing or lacking the Rnq1 protein which forms the [RNQ+] prion, providing new clues to the as yet unresolved normal biological function of this prion-forming protein. This investigation further emphasizes the utility of HPTLC-densitometry to empirically determine the effects of prions and other presumed innocuous gene deletions on lipid content in yeast, and we expect that additional analyses will continue to resolve the physiological effects of prion infection.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Using High Performance Thin Layer Chromatography-Densitometry to Study the Influence of the Prion [RNQ+] and Its Determinant Prion Protein Rnq1 on Yeast Lipid Profiles

Sherma

Hines

2018

Separations

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“…According to Bui et al, HPTLC-densitometry is a reproducible and accurate method for quantifying lipids in yeast cells. [25,26,107] Thus, the influence of the prion [RNQ + ] and its determinant prion protein Rnq1 on yeast lipid profiles (Saccharomyces cerevisiae) was studied. Using different development schemes, free sterols, FFA, TG, SterylE, methyl esters, Sq, PC, PE, PI were separated.…”

Section: Semi-quantave and Quantave Analysesmentioning

confidence: 99%

Scanning densitometry and mass spectrometry for HPTLC analysis of lipids: The last 10 years

Cebolla

Jarne

Vela

et al. 2021

Journal of Liquid Chromatography & Related Technologies

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This work is a review on HPTLC contribuon to lipid analysis in complex matrices, in the period from 2010 to now, lapse of me where hyphenaon with other techniques, especially Mass Spectrometry, has experienced an important growth.Scanning densitometry (SD)-UV/FL of separated lipids, as the core of detecon and centerpiece for hyphenaon, and its coupling with MS using so ionizaon techniques (ESI, APCI, MALDI, DESI and others), are the central axis of this work. The occasional intercalaon in this coupling of an on-plate biological assay for eect-direct analysis (EDA) of lipids, as well as the combinaon of SD with radiodensitometry (RD) using isotopically labeled-lipids are also covered topics.The described techniques make possible to develop strategies for obtaining qualitave and dierent levels of quantave informaon, including untargeted lipids species.HPTLC of lipids has been used for: comparave purposes; ngerprinng; semi-quantave determinaon, idencaon of species; or quantave determinaon of a given individual lipid. Goals in molecular biology and biochemistry-related samples have been: preparave isolaons; control of purity; vericaon of metabolic products; on-plate biological assays; proles in cells; analysis of products from the cellular metabolism; measurement of enzymac acvies; monitoring lipid transport across membranes or at biological interfaces; or monitoring lipid dynamics.

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Thin-Layer Chromatography and Mass Spectrometry for the Analysis of Lipids

Tuzimski

Sherma

2016

Encyclopedia of Lipidomics

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Thin-layer chromatographic analysis of lipids and lipophilic pigments in snails

Cited by 4 publications

References 45 publications

Using High Performance Thin Layer Chromatography-Densitometry to Study the Influence of the Prion [RNQ+] and Its Determinant Prion Protein Rnq1 on Yeast Lipid Profiles

Using High Performance Thin Layer Chromatography-Densitometry to Study the Influence of the Prion [RNQ+] and Its Determinant Prion Protein Rnq1 on Yeast Lipid Profiles

Scanning densitometry and mass spectrometry for HPTLC analysis of lipids: The last 10 years

Thin-Layer Chromatography and Mass Spectrometry for the Analysis of Lipids

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