Dataset Information

Quantitative structural characterization of phosphatidylinositol phosphates from biological samples.

ABSTRACT: The aspects of cellular metabolism controlled by phosphatidylinositol phosphates (PtdInsPs) have been broadly expanded, and these phospholipids have drawn tremendous attention as pleiotropic signaling molecules. PtdInsPs analysis using LC/MS/MS has remained challenging due to the strong hydrophilicity of these lipids. Multiple reaction monitoring (MRM) or a neutral loss scan has been performed to quantitatively measure PtdInsPs after chemical derivatization on the phosphate groups of inositol moieties. Only predefined PtdInsPs can be measured in MRM mode, and fatty acyl compositions of sn-1 and sn-2 positions of PtdInsPs cannot be obtained from a neutral loss scan. In our present study, we developed a simple LC/MS/MS method for structural identification of sn-1 and sn-2 fatty acids of PtdInsPs and their relative quantitation. Precursor ion scans of sn-1 monoacylglycerols (MAGs) of PtdInsPs provided structural information about the lipids, and ammonium adduction enhanced signal intensities of PtdInsPs. The relative amount of observed PtdInsPs in biological samples could be compared using chromatographic peak areas from the neutral loss scans. Using precursor ion scans of sn-1 MAG and neutral loss scans of headgroups, major PtdInsPs in cells and tissues were successfully identified with structural information of sn-1 and sn-2 fatty acids, and their relative amounts in different samples were compared.

SUBMITTER: Kim SH

PROVIDER: S-EPMC5282945 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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Quantitative structural characterization of phosphatidylinositol phosphates from biological samples.

Kim Su Hee SH Song Ha Eun HE Kim Su Jung SJ Woo Dong Cheol DC Chang Suhwan S Choi Woo Gyun WG Kim Mi Jeong MJ Back Sung Hoon SH Yoo Hyun Ju HJ

Journal of lipid research 20161209 2

The aspects of cellular metabolism controlled by phosphatidylinositol phosphates (PtdInsPs) have been broadly expanded, and these phospholipids have drawn tremendous attention as pleiotropic signaling molecules. PtdInsPs analysis using LC/MS/MS has remained challenging due to the strong hydrophilicity of these lipids. Multiple reaction monitoring (MRM) or a neutral loss scan has been performed to quantitatively measure PtdInsPs after chemical derivatization on the phosphate groups of inositol mo ...[more]

PMID: 27940482

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Project description:Graphical abstract Assessment of vitamin A concentrations in biological specimens or functional indicators of vitamin A status (severity of night blindness or lack thereof) require field-friendly methods for assessment to expand coverage, particularly in low-income settings. In this review, portable devices for measurement were compared with reference methods across key criteria. For example, portable devices to determine vitamin A concentrations in blood or milk samples were catalogued and compared with the reference method of high performance liquid chromatography, and a portable dark adaptometer to assess vision was compared against the reference of a Goldmann-Weekers dark adaptometer. Highlights • Vitamin A deficiency is a major global health issue leading to poor health outcomes.• Vitamin A assessment usually requires a centralized laboratory and equipment.• Portable methods to assess vitamin A may overcome the limitations of laboratory-based testing.• Blood, milk, and eye function may be used for vitamin A status measurement. Background We catalog and summarize evidence of the analytical performance of portable quantitative and semi-quantitative devices for the assessment of vitamin A status and vitamin A deficiency (VAD) in various biological samples—including whole blood, plasma, serum, and milk—in addition to VAD determination by functional indicators such as pupillary response. Methods We searched the literature for published research articles, patents, and information from manufacturers of mobile devices, particularly those appropriate for low-resource settings. The included devices were required to be portable (lightweight and ideally not needing a power outlet) and to measure vitamin A as well as define VAD. Eligible studies compared a portable device to a reference standard of high-performance liquid chromatography for blood and milk, or a Goldmann-Weekers dark adaptometer for eyes/vision. Where available, identified devices were compared with reference methods across several performance criteria. When possible, we compared the device’s performance reported in published studies against the stated performance criteria from the manufacturers’ websites. Results We catalogued 25 portable devices for measuring vitamin A and/or VAD via biological samples. We also identified 18 comparison studies (plus associated reports) assessing nine methods: the iCheck Fluoro, iCheck Carotene, CRAFTi, Tidbit with or without the HYPER filtration system, custom field-friendly immunoassays, and microfluidic assays for blood; the iCheck Fluoro and iCheck Carotene for milk; and the Scotopic Sensitivity Tester-1 for eye function. Conclusions The iCheck Fluoro and iCheck Carotene are commercially available for use and are acceptable for measuring vitamin A in blood and milk samples, according to the available validation data. Many of the other identified devices, including other portable fluorometers, photometers, immunoassays, microfluidics-based devices, and dark adaptometers, were proofs of concept and not yet commercially available. Furthermore, none of these other devices included manufacturer-described device performance criteria to compare with descriptions from experimental studies. Several gaps remain, including studies comparing the other portable devices against a reference standard, particularly for functional indicators of vitamin A status/deficiency; available manufacturer-reported device performance criteria against which to compare future results of investigations; and more comprehensive reporting of validation metrics including sensitivity, specificity, precision, and Bland-Altman analysis.

Dataset Information

Quantitative structural characterization of phosphatidylinositol phosphates from biological samples.

Publications

Quantitative structural characterization of phosphatidylinositol phosphates from biological samples.

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