A comparative study of the phospholipid profiles of guinea pig cardiac muscle and bullfrog cardiac and thigh skeletal muscle, and their in-vitro differential deacylation by endogenous phospholipases. Thin layer chromatographic and densitometric analysis

Abstract

Several detailed reviews on cellular phospholipase A2 (cPLA2) have been published, however the role of cPLA2 in cell function remains unclear. Continued substantive efforts are required to elucidate a broader understanding of the in vivo functionality for all endogenous phospholipases and an explanation for the need for such diverse phospholipids in various biological membranes, which are not yet at hand. Recently, we have reported on the preferential deacylation of cardiolipin (CL) of various mammalian myocardia by an endogenous phospholipase (PLA1 and/or PLA2), following in vitro incubation at pH 7.4 and 38°C for 60 minutes of whole tissue homogenate (as source of phospholipids and phospholipases) producing monolysocardiolipin (MLCL) and concurrent reduction of CL. In contrast whole tissue homogenate of rabbit spleen treated similarly selectively deacylated ethanolamine plasmalogen (PE) producing lyso alkenyl PE. In the present study, whole tissue homogenate of bullfrog cardiac muscle and thigh skeletal muscle, treated similarly revealed the following data: Bullfrog cardiac muscle slightly deacylated CL producing MLCL, Thigh skeletal muscle shows very minute amount of CL and no deacylation products following in vitro incubation, PE plasmalogen is the only alkenyl species of both tissues, Thigh muscle has very minute amount of Sphingomyelin. These data raised some questions pertaining to the Lipolytic capabilities of diverse tissues from different species of animals and will be discussed in details.

Optimized extraction of phospholipids and lysophospholipids for nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry

“…Among the different subclasses of lipids, lysophospholipids (LPLs) and PLs are often recognized as potential biomarkers of human diseases such as breast cancer, [5][6][7] ovarian cancer, 8,9 and prostate cancer. 10,11 Intact PLs have been analyzed with simple thin layer chromatography (TLC) in conjunction with detection methods such as densitometry 12 or matrix-assisted laser desorption and ionization/time of flight mass spectrometry (MALDI-TOF/MS). 13 With rapid advances in the MS technique due to the adoption of electrospray ionization (ESI), the direct ESI-MS of intact PLs [14][15][16][17] or liquid chromatography (LC) coupled with ESI-MS [18][19][20][21] has expanded the scope of lipidomics toward efforts to discover potential biomarkers of adult diseases.…”

Profiling of human urinary phospholipids by nanoflow liquid chromatography/tandem mass spectrometry

“…Conventionally, intact phospholipids are studied by analytical techniques such as the simple thin layer chromatography (TLC) with various detection ways: phosphorous detection method, 4,5 matrix-assisted laser desorption and ionization time-of flight mass spectrometry (MALDI-TOF MS), 6,7 and densitometry. 8,9 High performance liquid chromatography (HPLC) is utilized with UV detection, 10,11 and diode array detection (DAD). 12 Recent advances in mass spectrometry (MS) coupled with electrospray ionization (ESI) have allowed the rapid and highly sensitive analysis of PLs [13][14][15] as well as quantitative analysis, 16 but ESI-MS alone can not bypass the ion suppression effect caused by the complexity of PL molecules.…”

A Comparative Study Concerning the Image Analysis in Thin Layer Chromatography of Fluorescent Compounds

“…TLC was a rapid, simple, and inexpensive method providing the possibility of processing multiple samples in parallel with a high throughput. [4,5] In spite of all these advantages offered by the TLC, the method had a reduced sensitivity and limited delectability and, therefore, it was only used in qualitative analysis. However, TLC became a quantitative method when it was coupled with slit scanning densitometers and later with charged-coupled devices (CCD) which considerably improved the accuracy of the method.…”