Some practical aspects of thin‐layer chromatography of lipids

Pelick, Nicholas; Wilson, T. L.; Miller, M. E.; Angeloni, F. M.; Steim, Joseph M.

doi:10.1007/bf02635574

Cited by 31 publications

(4 citation statements)

References 22 publications

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“…The fact that the glucose to acyl ester to glycerol ratios for the two lipids were compatible with the identification of the lipids by chromatography indicated that the anthrone reaction was a good measure of the lipid glucose. The two glucolipids were easily separated by TLC in the chloroform-methanol-acetic acid solvent (36). The areas near RF values of 0.66 (diglucosyldiglyceride) and 0.95 (monoglucosyldiglyceride) were scraped off and eluted.…”

Section: Aqueousmentioning

confidence: 99%

Extraction, Characterization, and Cellular Localization of the Lipids of Staphylococcus aureus

1967

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Satisfactory extraction and assay procedures have been developed for the lipids of Staphylococcus aureus. The following lipids have been characterized in detail: the vitamin K2, which is shown to exist as isoprenologues with side chains of 35, 40, and 45 carbon atoms; monoglucosyldiglyceride and diglucosyldiglyceride, which account for all the carbohydrate in the lipid extracts; the lysyl ester of phosphatidyl glycerol, phosphatidyl glycerol, and cardiolipin, which account for 98% of the phosphate in the lipid extract. The extraction procedure removes 98% of the total bacterial fatty acids. Acidification of the medium before harvest and refluxing in isopropanol are critical in the extraction procedure for the maximal recovery of lysyl-phosphatidyl glycerol and the glucolipids. The lipids have been shown to be a part of the same membrane as the respiratory pigments. Staphylococcus aureus has the capacity to grow anaerobically with the energy produced from the glycolytic pathway. While growing anaerobically, the enzymes of the Krebs cycle and the membrane-bound electron transport system are repressed (6, 46). Addition of oxygen to the growing culture causes the induction of the membrane-bound electron transport system (12). This formation of the multienzyme respiratory system is coordinated with the synthesis of neutral lipid, glycolipid, phospholipids, and other components. The function of the electron transport system requires structural integrity, since any treatment with organic solvents destroys the activity. To study the process of membrane formation, the components of the membrane must be identified and the techniques for their assay developed. This paper gives assay procedures for the lipids and establishes that these lipids are a part of the respiratory membrane complex. Staphylococcus aureus has been shown to have a membrane-bound electron transport system consisting of cytochromes bi and a and cytochrome oxidase o (47). These bacteria contain a vitamin K2 derivative which is formed when the bacteria are grown with aeration (3). A glycolipid, diglucosyl diglyceride, contains the glucose extractable with lipid solvents (37). Lysyl-phosphatidyl glycerol (LPG), phosphatidyl glycerol (PG), and cardiolipin are the principal phospho-MATERIALS AND METHODS Growth of bacteria. Staphylococcus aureus strain U-71 was provided by J. N. Baldwin. The medium used to grow the bacteria was either the semisynthetic medium containing acid-hydrolyzed casein (22) or this medium supplemented with 0.2% (w/v) Difco yeast extract. Supplements were added as recommended (14). The final concentrations were: 10 mm acetate, 1 mM uracil, 1 mn xanthine, 1 mim adenine, and 15 mm glucose. The tryptophan, iron, vitamins, and glucose were added after autoclaving the medium. The bacteria were preserved in 15% (v/v) glycerol in the growth medium at-60 C. Purity of the culture was checked by Gram stain, colonial morphology, and phase-contrast microscopy. Dry weight. The relationship between dry weight of the bacteria and absorbance of the bacte...

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Section: Aqueousmentioning

confidence: 99%

Extraction, Characterization, and Cellular Localization of the Lipids of Staphylococcus aureus

1967

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“…The following paper chromatography systems were employed. Intact lipids were chromatographed on silicic acid impregnated paper (Whatman SG81) developed with the following solvent systems: (A) diisobutyl ketone-acetic acid-water (40:25:5, v/v) (Marinetti et al, 1957), (B) chloroform-methanol-water (65:25:4, v/v) (Pelick et al, 1965), (C) chloroform-methanol-water (45:10:1, v/v) (Pelick et al, 1965). Water-soluble acid and mild alkaline hydrolysis products of the lipids were chromatographed on Whatman No.…”

Section: Methodsmentioning

confidence: 99%

The metabolism of glyceride glycolipids. II. Biosynthesis of monogalactosyl diglyceride from uridine diphosphate galactose and diglyceride in brain

Wenger¹,

Petitpas²,

Pieringer³

1968

Biochemistry

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“…In addition, if proper care isn't exercised, 1 Present address, USDA, ARS, Human Nutrition Research Division, Food Quality and Use Laboratory, Room 316, Center Bldg., Agricultural Research Center, Beltsville, Md. 20705 separation by TLC can lead to oxidative losses of highly unsaturated fatty acids and recovery of the phospholipids may be incomplete (1,2). Phospholipids have been removed from lipid extracts by adsorption on activated silicic acid (3, 4) and free fatty acids have been removed from lipid extracts by adsorption on anion exchange resins (5).…”

Section: Separation Of Muscle Lipids Into Classes By Nonchromatograph...mentioning

confidence: 99%

Spark source mass spectrometric measurements of dopants of known concentrations in gallium phosphide

Ahearn¹,

Trumbore²,

Frosch³

et al. 1967

Anal. Chem.

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Some practical aspects of thin‐layer chromatography of lipids

Cited by 31 publications

References 22 publications

Extraction, Characterization, and Cellular Localization of the Lipids of Staphylococcus aureus

Extraction, Characterization, and Cellular Localization of the Lipids of Staphylococcus aureus

The metabolism of glyceride glycolipids. II. Biosynthesis of monogalactosyl diglyceride from uridine diphosphate galactose and diglyceride in brain

Spark source mass spectrometric measurements of dopants of known concentrations in gallium phosphide

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