Separation of synthetic phosphatidylcholine molecular species by high-performance liquid chromatography on a C8 column

Lin, Jiann‐Tsyh; McKeon, Thomas A.; Woodruff, C. L.; Singleton, John

doi:10.1016/s0021-9673(98)00638-4

Cited by 18 publications

(24 citation statements)

References 14 publications

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“…n--9 elutes before n--12, Figure 2a). This observation is consistent with previous reports of separation of double bond positional isomers and evidence from new column technologies suggests further improvements in isomer separation can be achieved [17,19,18]. Full structure elucidation of complex lipids however, also requires separation and/or mass spectral differentiation of isomers that differ in (i) the position of acyl chains on the glycerol backbone (i.e., sn--positional isomers) and the stereochemistry about the double bond(s) (i.e., cis and trans).…”

Section: Discussionsupporting

confidence: 93%

“…Prior observation of the behaviour of unsaturated phosphatidylcholines on reversed phase columns suggests that the peaks eluting at 49.0 and 50.4 min correspond to the isomers PC 18:1(n--9)/18:1(n--9) and PC 18:1(n--12)/18:1(n--12), respectively [17,19,18]. Integration of OzID scans obtained across each of the chromatographic peaks gave the OzID mass spectra shown in bonds closer to the methyl terminus of lipid acyl chains has previously been noted [24].…”

Section: [Figure 2]mentioning

confidence: 98%

“…In contrast, analytical separations of phospholipid double bond positional isomers -where the acyl chains are themselves isomeric -by reversed--phase LC have proven more challenging. Prepared mixtures of two synthetic phosphatidylcholine double bond positional isomers, PC 18:1(n--9)/18:1(n--9) and PC 18:1(n--12)/18:1(n--12), have been successfully separated by reversed--phase LC using a C 8 column and, more recently, using charged surface hybrid C 18 columns [17,18]. While these analytical demonstrations are important, very few studies have differentiated between endogenous double bond positional isomers in complex biological extracts.…”

Section: Introductionmentioning

confidence: 99%

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Combining liquid chromatography with ozone-induced dissociation for the separation and identification of phosphatidylcholine double bond isomers

et al. 2015

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. Combining liquid chromatography with ozone-induced dissociation for the separation and identification of phosphatidylcholine double bond isomers. Analytical and Bioanalytical Chemistry, 407 (17), 5053-5064. Combining liquid chromatography with ozone-induced dissociation for the separation and identification of phosphatidylcholine double bond isomers AbstractRevealing the inherent molecular diversity of lipid biology requires advanced analytical technologies. Distinguishing phospholipids that differ in the position(s) of carbon-carbon double bonds within their acyl chains presents a particular challenge because of their similar chromatographic and mass spectral behaviours. Here-for the first time-we combine reversed-phase liquid chromatography for separation of isomeric phospholipids with on-line mass spectral analysis by ozone-induced dissociation (OzID) for unambiguous double bond position assignment. The customised tandem linear ion-trap mass spectrometer used in our study is capable of acquiring OzID scans on a chromatographic timescale. Resolving the contributions of isomeric lipids that are indistinguishable based on conventional mass spectral analysis is achieved using the combination of liquid chromatography and OzID. Application of this method to the analysis of simple (egg yolk) and more complex (sheep brain) extracts reveals significant populations of the phosphatidylcholine PC 16:0_18:1(n−7) alongside the expected PC 16:0_18:1(n−9) isomer.

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Section: Discussionsupporting

confidence: 93%

Section: [Figure 2]mentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Combining liquid chromatography with ozone-induced dissociation for the separation and identification of phosphatidylcholine double bond isomers

et al. 2015

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“…(7). The relative retention times (RRT) of the molecular species of PC incorporating [ 14 C]laurate were estimated by normalization to the retention time of PO-PC (32.1 min) reported earlier(7).…”

mentioning

confidence: 99%

“…The relative retention times (RRT) of the molecular species of PC incorporating [ 14 C]laurate were estimated by normalization to the retention time of PO-PC (32.1 min) reported earlier(7). The RRT of these commercially unavail-able PC were: RLa-PC (15.1 min), LnLa-PC (18.8 min), LLa-PC (21.3 min), PLa-PC (23.1 min), OLa-PC (24.3 min), and SLa-PC (27.3 min).…”

mentioning

confidence: 99%

Incorporation of laurate and hydroxylaurate into phosphatidylcholines and acylglycerols in castor microsomes

Lin

Wani

et al. 2005

J Americ Oil Chem Soc

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Because castor produces oil with a high content of hydroxyl FA (90% ricinoleate), we were interested in determining the flexibility of castor seed microsomes in incorporating other hydroxyl FA into castor oil. To this end, we incubated the [ 14 C]-labeled 12:0 FA laurate (La), 11-hydroxylaurate, and 12-hydroxylaurate with castor microsomes that were capable of synthesizing castor oil. The molecular species of PC and acylglycerols (AG) incorporating these nonendogenous FA of castor were identified by reversed-phase C 8 and C 18 HPLC, respectively. [ 14 C]Laurate was incorporated into the molecular species of PC and AG at levels of 10 and 4%, respectively, that of [ 14 C]ricinoleate. Similar to those from the incorporation of six [ 14 C]FA reported previously [ricinoleate (R), oleate (O), linoleate (L), linolenate (Ln), stearate (S), and palmitate (P)], the molecular species of PC incorporating [ 14 C]laureate were LLa-PC > PLa-PC > OLa-PC > LnLa-PC > SLa-PC > RLa-PC. The molecular species of AG incorporating [ 14 C]laurate were RRLa > LaLa > RLa > RLLa > ROLa > LOLa > LLLa > LLa > LLnLa > RSLa > OOLa. The retention times for lipids incorporating laurate were similar to those of lipids incorporating linolenate, because the equivalent carbon numbers of laurate and linolenate are the same. Relative retention times of the molecular species of PC and AG containing laurate are also reported here. The incorporation of 11-hydroxylaurate and 12-hydroxylaurate into PC and AG was not detected. Paper no. J10984 in JAOCS 82, 495-499 (2005).The presence of a hydroxyl group on ricinoleate (12-hydroxyoleate, C 18 long-chain FA) underlies many industrial uses such as the manufacture of lithium grease, plastics, coatings, and cosmetics. Castor oil contains 90% of its FA as ricinoleate (1) and is the only commercial source of ricinoleate. We have recently studied the incorporation of six endogenous long-chain FA-ricinoleate (R), stearate (S), oleate (O), linoleate (L), linolenate (Ln), and palmitate (P)-into PC and acylglycerols (AG) in castor microsomes (2,3). Among the six FA, ricinoleate was incorporated into TAG most effectively. Since the position of the hydroxyl group on ricinoleate (C 18 ) is the same as on 12-hydroxylaurate (C 12 ), castor microsomal incorporation of radiolabeled laurate (La), 11-hydroxylaurate, and 12-hydroxylaurate into PC and AG was of interest to determine the flexibility of castor in producing other hydroxyl FA. Laurate is mainly found in lauric oils such as coconut and palm kernel and is used in food products and in the manufacture of soaps and detergents. The mediumchain hydroxyl FA 11-hydroxylauric acid and 12-hydroxylauric acid can be used in the manufacture of polymers. EXPERIMENTAL PROCEDURESMicrosomal incubation. Microsomes from castor bean were prepared as described previously, and microsomal incubations were scaled up 10 times with the addition of CoA-SH to generate suitable amounts of material for analysis (4,5). The incubation mixture, in a total volume of 10 mL, included sodium phosphate buf...

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Reversed-Phase Liquid Chromatography Mass Spectrometry (RP-LC-MS) in Lipidomics

t’Kindt¹,

Sandra²,

Sandra³

2016

Encyclopedia of Lipidomics

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Separation of synthetic phosphatidylcholine molecular species by high-performance liquid chromatography on a C8 column

Cited by 18 publications

References 14 publications

Combining liquid chromatography with ozone-induced dissociation for the separation and identification of phosphatidylcholine double bond isomers

Combining liquid chromatography with ozone-induced dissociation for the separation and identification of phosphatidylcholine double bond isomers

Incorporation of laurate and hydroxylaurate into phosphatidylcholines and acylglycerols in castor microsomes

Reversed-Phase Liquid Chromatography Mass Spectrometry (RP-LC-MS) in Lipidomics

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