Determination of phospholipids by a combined liquid chromatograph-automated phosphorus analyzer

Kaitaranta, Jukka K.; Bessman, Samuel P.

doi:10.1021/ac00231a023

Cited by 49 publications

(5 citation statements)

References 17 publications

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“…Pollen PLs were also subfractionated by HPLC (Kaitaranta and Bessman, 1981). They were loaded on a semipreparative HPLC column (15 cm length, 9.4 mm diameter, Spherisorb S5W; Merck, Darmstadt, Germany) connected to a Spectra Physics isochrom isocratic pump.…”

Section: Pl Purificationmentioning

confidence: 99%

Decreased Membrane Integrity in Aging Typha latifolia L.Pollen (Accumulation of Lysolipids and Free Fatty Acids)

Bilsen

Hoekstra

1993

Plant Physiol.

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Aging of cattail (Typha latifolia 1.) pollen was studied at 24°C under conditions of 40 and 75% relative humidity (RH). The decline of viability coincides with increased leakage at imbibition; both processes develop much faster at the higher humidity condition. During aging phospholipids are deesterified and free fatty acids (FFAs) and lysophospholipids (LPLs) accumulate, again, much more rapidly at 75% RH than at 40% RH. The fatty acid composition of the remaining phospholipids hardly changes during aging, which suggests limited involvement of lipid peroxidation in the degradation process. Tests with phospholipase A2 revealed that the saturated fatty acids occur at the sn-1 position of the glycerol backbone of the phospholipids. The fatty acid composition of the LPLs is similar to that of the phospholipids from which they were formed, indicating that the deesterification occurs at random. This favors involvement of free radicals instead of phospholipases i n the deesterification process. Liposome studies were carried out to characterize components i n the lipid fraction that might account for the leakage associated with aging. Entrapped carboxyfluorescein leaked much more from liposomes when they were partly made up from total lipids from aged pollen than from nonaged pollen.The components causing the leakage were found i n both the polar and the neutral lipid fractions. Further purification and subsequent interchanging of the FFAs and LPLs between extracts from aged and nonaged pollen revealed that in neutral lipid extracts the FFAs are entirely responsible for the leakage, whereas i n the phospholipid fraction the LPLs are largely responsible for the leakage. The leakage from the liposomes i s not caused by fusion. We suggest that the observed loss of viability and increased leakage during aging are due to the nonenzymic accumulation of FFAs and LPLs in the pollen membranes.

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Section: Pl Purificationmentioning

confidence: 99%

Decreased Membrane Integrity in Aging Typha latifolia L.Pollen (Accumulation of Lysolipids and Free Fatty Acids)

Bilsen

Hoekstra

1993

Plant Physiol.

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“…at a flow rate of 0-4 ml min-' (Kaitaranta & Bessman, 1981). The purified phospholipid was then separated into molecular species by reverse-phase HPLC using Spherisorb ODS I1 5 pm (4.6 x 150 mm) and a UV detector at 205 nm.…”

Section: Lipid Extractionmentioning

confidence: 99%

Positional distribution of fatty acids, and molecular species of polar lipids, in the diatom Phaeodactylum tricornutum

Yongmanitchai

Ward

1993

Journal of General Microbiology

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“…First, silica gel HPLC was performed in isopropanolln-hexanelwater 70:20:10 (v/v/v), and PC and SM were isocratically eluted and found to overlap [17]. Then SM was isolated on the same column using acetonitrile/methanol/water 70:20:10 (v/v/v) [18].…”

Section: Analysis Of the Molecular Species Of Cmh Cdh And Smmentioning

confidence: 99%

Clear differences in ceramide metabolism between glycosphingolipids and sphingomyelin in a human promyelocytic leukemia cell line HL‐60 stimulated by a differentiation inducer

1995

FEBS Letters

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Although the ceramide components of both glycosphingolipids (GSLs) and sphingomyelin (SM) in HL-60 cells were identical, the molecular species of the ceramides preferentially used in biosynthesis were quite different in GSLs and SM. When HL-60 cells were stimulated to differentiate into macrophage-like cells by phorbol ester after their sphingolipids had been metabolically labeled with L-13-14C]serine to saturation point, marked changes in the radioactivities of the ceramide residues were observed in GSLs, showing the activation of a biosynthetic pathway of ganglioside GM3. No significant changes were, however, observed in the ceramide residues of SM. These results indicate that it is necessary to consider the overall metabolism of ceramides, including their origin, when investigating the functions of ceramides in signal transduction systems.tion of the possible role of GSLs, even though both GSLs and SM contain ceramide. To clarify-the physiological significance of sphingolipids, it is important to study the overall cellular stimulus-induced metabolism of GSLs and SM. However, only a few attempts have so far been made to investigate the relationship between their metabolic pathways [4,5].In the present study, we first established a simple method which enabled simultaneous analysis of GSLs and SM in the total lipid fraction of cultured cells by two-dimensional HPTLC. Early changes in sphingolipid metabolism, including GSLs and SM, during the macrophage-like differentiation of HL-60 cells [6,7] were then analyzed to estimate the physiological significance of sphingolipids in the differentiation process.

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Determination of phospholipids by a combined liquid chromatograph-automated phosphorus analyzer

Cited by 49 publications

References 17 publications

Decreased Membrane Integrity in Aging Typha latifolia L.Pollen (Accumulation of Lysolipids and Free Fatty Acids)

Decreased Membrane Integrity in Aging Typha latifolia L.Pollen (Accumulation of Lysolipids and Free Fatty Acids)

Positional distribution of fatty acids, and molecular species of polar lipids, in the diatom Phaeodactylum tricornutum

Clear differences in ceramide metabolism between glycosphingolipids and sphingomyelin in a human promyelocytic leukemia cell line HL‐60 stimulated by a differentiation inducer

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