Using precursor ion scan of 184 with liquid chromatography-electrospray ionization-tandem mass spectrometry for concentration normalization in cellular lipidomic studies

Hsi Chun Chao, Guan Yuan Chen, Lih Ching Hsu, Hsiao Wei Liao, Sin Yu Yang, San Yuan Wang, Yu Liang Li, Sung Chun Tang, Yufeng Jane Tseng, Ching Hua Kuo

研究成果: 雜誌貢獻文章

6 引文 (Scopus)

摘要

Cellular lipidomic studies have been favored approaches in many biomedical research areas. To provide fair comparisons of the studied cells, it is essential to perform normalization of the determined concentration before lipidomic analysis. This study proposed a cellular lipidomic normalization method by measuring the phosphatidylcholine (PC) and sphingomyelin (SM) contents in cell extracts. To provide efficient analysis of PC and SM in cell extracts, flow injection analysis-electrospray ionization-tandem mass spectrometry (FIA-ESI-MS/MS) with a precursor ion scan (PIS) of m/z 184 was used, and the parameters affecting the performance of the method were optimized. Good linearity could be observed between the cell extract dilution factor and the reciprocal of the total ion chromatogram (TIC) area in the PIS of m/z 184 within the dilution range of 1- to 16-fold (R2 = 0.998). The calibration curve could be used for concentration adjustment of the unknown concentration of a cell extract. The intraday and intermediate precisions were below 10%. The accuracy ranged from 93.0% to 105.6%. The performance of the new normalization method was evaluated using different numbers of HCT-116 cells. Sphingosine, ceramide (d18:1/18:0), SM (d18:1/18:0) and PC (16:1/18:0) were selected as the representative test lipid species, and the results showed that the peak areas of each lipid species obtained from different cell numbers were within a 20% variation after normalization. Finally, the PIS of 184 normalization method was applied to study ischemia-induced neuron injury using oxygen and glucose deprivation (OGD) on primary neuronal cultured cells. Our results showed that the PIS of 184 normalization method is an efficient and effective approach for concentration normalization in cellular lipidomic studies.
原文英語
頁(從 - 到)68-77
頁數10
期刊Analytica Chimica Acta
971
DOIs
出版狀態已發佈 - 六月 8 2017
對外發佈Yes

指紋

Electrospray ionization
Electrospray Ionization Mass Spectrometry
Liquid chromatography
Tandem Mass Spectrometry
Liquid Chromatography
Mass spectrometry
liquid chromatography
Cell Extracts
ionization
mass spectrometry
Sphingomyelins
Ions
Phosphatidylcholines
ion
Dilution
Cells
Flow Injection Analysis
Lipids
Sphingosine
dilution

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

引用此文

Using precursor ion scan of 184 with liquid chromatography-electrospray ionization-tandem mass spectrometry for concentration normalization in cellular lipidomic studies. / Chao, Hsi Chun; Chen, Guan Yuan; Hsu, Lih Ching; Liao, Hsiao Wei; Yang, Sin Yu; Wang, San Yuan; Li, Yu Liang; Tang, Sung Chun; Tseng, Yufeng Jane; Kuo, Ching Hua.

於: Analytica Chimica Acta, 卷 971, 08.06.2017, p. 68-77.

研究成果: 雜誌貢獻文章

Chao, Hsi Chun ; Chen, Guan Yuan ; Hsu, Lih Ching ; Liao, Hsiao Wei ; Yang, Sin Yu ; Wang, San Yuan ; Li, Yu Liang ; Tang, Sung Chun ; Tseng, Yufeng Jane ; Kuo, Ching Hua. / Using precursor ion scan of 184 with liquid chromatography-electrospray ionization-tandem mass spectrometry for concentration normalization in cellular lipidomic studies. 於: Analytica Chimica Acta. 2017 ; 卷 971. 頁 68-77.
@article{ea90be89e3194a489b37ff112ddcf562,
title = "Using precursor ion scan of 184 with liquid chromatography-electrospray ionization-tandem mass spectrometry for concentration normalization in cellular lipidomic studies",
abstract = "Cellular lipidomic studies have been favored approaches in many biomedical research areas. To provide fair comparisons of the studied cells, it is essential to perform normalization of the determined concentration before lipidomic analysis. This study proposed a cellular lipidomic normalization method by measuring the phosphatidylcholine (PC) and sphingomyelin (SM) contents in cell extracts. To provide efficient analysis of PC and SM in cell extracts, flow injection analysis-electrospray ionization-tandem mass spectrometry (FIA-ESI-MS/MS) with a precursor ion scan (PIS) of m/z 184 was used, and the parameters affecting the performance of the method were optimized. Good linearity could be observed between the cell extract dilution factor and the reciprocal of the total ion chromatogram (TIC) area in the PIS of m/z 184 within the dilution range of 1- to 16-fold (R2 = 0.998). The calibration curve could be used for concentration adjustment of the unknown concentration of a cell extract. The intraday and intermediate precisions were below 10{\%}. The accuracy ranged from 93.0{\%} to 105.6{\%}. The performance of the new normalization method was evaluated using different numbers of HCT-116 cells. Sphingosine, ceramide (d18:1/18:0), SM (d18:1/18:0) and PC (16:1/18:0) were selected as the representative test lipid species, and the results showed that the peak areas of each lipid species obtained from different cell numbers were within a 20{\%} variation after normalization. Finally, the PIS of 184 normalization method was applied to study ischemia-induced neuron injury using oxygen and glucose deprivation (OGD) on primary neuronal cultured cells. Our results showed that the PIS of 184 normalization method is an efficient and effective approach for concentration normalization in cellular lipidomic studies.",
keywords = "Cellular lipidomics, Flow injection analysis-electrospray ionization-tandem mass spectrometry, Neuron damage, Normalization, Oxygen and glucose deprivation, Precursor ion scan of 184",
author = "Chao, {Hsi Chun} and Chen, {Guan Yuan} and Hsu, {Lih Ching} and Liao, {Hsiao Wei} and Yang, {Sin Yu} and Wang, {San Yuan} and Li, {Yu Liang} and Tang, {Sung Chun} and Tseng, {Yufeng Jane} and Kuo, {Ching Hua}",
year = "2017",
month = "6",
day = "8",
doi = "10.1016/j.aca.2017.03.033",
language = "English",
volume = "971",
pages = "68--77",
journal = "Analytica Chimica Acta",
issn = "0003-2670",
publisher = "Elsevier",

}

TY - JOUR

T1 - Using precursor ion scan of 184 with liquid chromatography-electrospray ionization-tandem mass spectrometry for concentration normalization in cellular lipidomic studies

AU - Chao, Hsi Chun

AU - Chen, Guan Yuan

AU - Hsu, Lih Ching

AU - Liao, Hsiao Wei

AU - Yang, Sin Yu

AU - Wang, San Yuan

AU - Li, Yu Liang

AU - Tang, Sung Chun

AU - Tseng, Yufeng Jane

AU - Kuo, Ching Hua

PY - 2017/6/8

Y1 - 2017/6/8

N2 - Cellular lipidomic studies have been favored approaches in many biomedical research areas. To provide fair comparisons of the studied cells, it is essential to perform normalization of the determined concentration before lipidomic analysis. This study proposed a cellular lipidomic normalization method by measuring the phosphatidylcholine (PC) and sphingomyelin (SM) contents in cell extracts. To provide efficient analysis of PC and SM in cell extracts, flow injection analysis-electrospray ionization-tandem mass spectrometry (FIA-ESI-MS/MS) with a precursor ion scan (PIS) of m/z 184 was used, and the parameters affecting the performance of the method were optimized. Good linearity could be observed between the cell extract dilution factor and the reciprocal of the total ion chromatogram (TIC) area in the PIS of m/z 184 within the dilution range of 1- to 16-fold (R2 = 0.998). The calibration curve could be used for concentration adjustment of the unknown concentration of a cell extract. The intraday and intermediate precisions were below 10%. The accuracy ranged from 93.0% to 105.6%. The performance of the new normalization method was evaluated using different numbers of HCT-116 cells. Sphingosine, ceramide (d18:1/18:0), SM (d18:1/18:0) and PC (16:1/18:0) were selected as the representative test lipid species, and the results showed that the peak areas of each lipid species obtained from different cell numbers were within a 20% variation after normalization. Finally, the PIS of 184 normalization method was applied to study ischemia-induced neuron injury using oxygen and glucose deprivation (OGD) on primary neuronal cultured cells. Our results showed that the PIS of 184 normalization method is an efficient and effective approach for concentration normalization in cellular lipidomic studies.

AB - Cellular lipidomic studies have been favored approaches in many biomedical research areas. To provide fair comparisons of the studied cells, it is essential to perform normalization of the determined concentration before lipidomic analysis. This study proposed a cellular lipidomic normalization method by measuring the phosphatidylcholine (PC) and sphingomyelin (SM) contents in cell extracts. To provide efficient analysis of PC and SM in cell extracts, flow injection analysis-electrospray ionization-tandem mass spectrometry (FIA-ESI-MS/MS) with a precursor ion scan (PIS) of m/z 184 was used, and the parameters affecting the performance of the method were optimized. Good linearity could be observed between the cell extract dilution factor and the reciprocal of the total ion chromatogram (TIC) area in the PIS of m/z 184 within the dilution range of 1- to 16-fold (R2 = 0.998). The calibration curve could be used for concentration adjustment of the unknown concentration of a cell extract. The intraday and intermediate precisions were below 10%. The accuracy ranged from 93.0% to 105.6%. The performance of the new normalization method was evaluated using different numbers of HCT-116 cells. Sphingosine, ceramide (d18:1/18:0), SM (d18:1/18:0) and PC (16:1/18:0) were selected as the representative test lipid species, and the results showed that the peak areas of each lipid species obtained from different cell numbers were within a 20% variation after normalization. Finally, the PIS of 184 normalization method was applied to study ischemia-induced neuron injury using oxygen and glucose deprivation (OGD) on primary neuronal cultured cells. Our results showed that the PIS of 184 normalization method is an efficient and effective approach for concentration normalization in cellular lipidomic studies.

KW - Cellular lipidomics

KW - Flow injection analysis-electrospray ionization-tandem mass spectrometry

KW - Neuron damage

KW - Normalization

KW - Oxygen and glucose deprivation

KW - Precursor ion scan of 184

UR - http://www.scopus.com/inward/record.url?scp=85016452485&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85016452485&partnerID=8YFLogxK

U2 - 10.1016/j.aca.2017.03.033

DO - 10.1016/j.aca.2017.03.033

M3 - Article

AN - SCOPUS:85016452485

VL - 971

SP - 68

EP - 77

JO - Analytica Chimica Acta

JF - Analytica Chimica Acta

SN - 0003-2670

ER -