Abstract
Amethod for quantitating phospholipids separated on thin layer chromatographic plates by computer-assisted photodensitometry is described. After development, the plates are stained with molibdic reagent and the image obtained is acquired as TIFF file in the memory of a personal computer. The color intensity of the single spots of the digitalized image is analyzed using a dedicated software.
Sensitivity and reproducibility are adequate for most of the needs of lipid chemist. When compared to conventional photodensitometric procedures, the present method offers the advantage of requiring a much cheaper hardware.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bartlett GR: Phosphorus assay in column chromatography. J Biol Chem 234: 466–468, 1959
van Vendhoven PP, Mannaerts GP: Inorganic and organic phosphate measurements in the nanomolar range. Anal Biochem 161: 45–48, 1987
Baykov AA, Evtushenko OA, Aveava SM: A malachite green procedure for ortophosphate determination and its use in alkaline phosphate-based enzyme immunoassay. Anal Biochem 171: 266–271, 1988
Nagata T, Poulsen LL, Ziegler DM: Estimation of lipid concentrations on thin layer plates by densitometry of transparent copies. Anal Biochem 171: 248–255, 1988
Stolywho A, Colin H, Guiochon G: Analysis of triglycerides in oils and fats by liquid chromatography with laser light scattering detector. Anal Chem 57: 1342–1354, 1985
Almog R, Anderson-Samsonoff C, Berns DS, Saulsbery R: A methodology for determination of phospholipids. Anal Biochem 188: 237–242, 1990
Rouser G, Kritchevski G, Yamamoto A. Column chromatographic and associated procedures for separation and determination of phosphatides and glycolipids. In: G. V. Marinetti. Lipid Chromatographic Analysis. Marcel Dekker, New York, Vol. 1, 1967, pp 99
Zwaal RFA, Roelofsen B, Comfurius P, van Deen LLM: Organization of phospholipids in human red cell membranes as detected by the action of various purified phospholipases. Biochim Biophys Acta 406: 83–96, 1975
Horrocks LA: The alk-1-enyl group content of mammalian myelin phosphoglycerides by two-dimensional thin layer chromatography. J Lipid Res 9: 469–472, 1968
Dittmer JD, Lester RL: A simple, specific spray for the detection of phospholipids on thin layer chromatograms. J Lipid Res 5: 126–133, 1964
Folch J, Lees M, Sloane-Stanley GH: A simple method for isolation and purification of total lipids for animal tissues. J Biol Chem 226: 497–509, 1957
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Vecchini, A., Chiaradia, E., Covalovo, S. et al. Quantitation of phospholipids on thin layer chromatographic plates using a desk-top scanner. Mol Cell Biochem 145, 25–28 (1995). https://doi.org/10.1007/BF00925709
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00925709