Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S]
Iso-octyl chain-hydroxylated oxysterols were determined in attomoles per 10,000 cells concentrations in 10,000–80,000 cultured pancreatic adenocarcinoma cells, using a sensitive, highly automated nano-LC-ESI-MS-based method. Identified oxysterols included 24S hydroxycholesterol (24S-OHC), 25 hydroxy...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-07-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520352585 |
| _version_ | 1819127481811075072 |
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| author | Hanne Roberg-Larsen Kaja Lund Tore Vehus Nina Solberg Caroline Vesterdal Dorna Misaghian Petter Angell Olsen Stefan Krauss Steven Ray Wilson Elsa Lundanes |
| author_facet | Hanne Roberg-Larsen Kaja Lund Tore Vehus Nina Solberg Caroline Vesterdal Dorna Misaghian Petter Angell Olsen Stefan Krauss Steven Ray Wilson Elsa Lundanes |
| author_sort | Hanne Roberg-Larsen |
| collection | DOAJ |
| description | Iso-octyl chain-hydroxylated oxysterols were determined in attomoles per 10,000 cells concentrations in 10,000–80,000 cultured pancreatic adenocarcinoma cells, using a sensitive, highly automated nano-LC-ESI-MS-based method. Identified oxysterols included 24S hydroxycholesterol (24S-OHC), 25 hydroxycholesterol (25-OHC), and 27 hydroxycholesterol (27-OHC), while 20S hydroxycholesterol and 22S hydroxycholesterol were not detected. Lower mass limit of quantification was 23 fg (65 amol) for 25-OHC and 27-OHC (100 times lower than our previous method) and 54 fg (135 amol) for 24S-OHC, after derivatization into Girard T hydrazones and online sample cleanup using simplified and robust automatic filtration and filter back flushing solid phase extraction LC/MS/MS. The instrument configuration was easily installed using a commercial nano-LC/MS system. Recoveries in spiked sample were 96, 97, and 77% for 24S-OHC, 25-OHC, and 27-OHC, with within- and between-day repeatabilities of 1–21#x0025; and 2–20#x0025; relative SD, respectively. The study demonstrates the potential of nano-LC in lipidomics/sterolomics. |
| first_indexed | 2024-12-22T08:12:37Z |
| format | Article |
| id | doaj.art-77b20f5d249949f682e91617898a85eb |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-22T08:12:37Z |
| publishDate | 2014-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-77b20f5d249949f682e91617898a85eb2022-12-21T18:32:59ZengElsevierJournal of Lipid Research0022-22752014-07-0155715311536Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S]Hanne Roberg-Larsen0Kaja Lund1Tore Vehus2Nina Solberg3Caroline Vesterdal4Dorna Misaghian5Petter Angell Olsen6Stefan Krauss7Steven Ray Wilson8Elsa Lundanes9Department of Chemistry, University of Oslo, Oslo, NorwayUnit for Cell Signaling, Cancer Stem Cell Innovation Centre (SFI-CAST), Oslo University Hospital, Rikshospitalet, Oslo, Norway; Affitech Research AS, Oslo, NorwayDepartment of Chemistry, University of Oslo, Oslo, NorwayUnit for Cell Signaling, Cancer Stem Cell Innovation Centre (SFI-CAST), Oslo University Hospital, Rikshospitalet, Oslo, NorwayDepartment of Chemistry, University of Oslo, Oslo, NorwayDepartment of Chemistry, University of Oslo, Oslo, NorwayUnit for Cell Signaling, Cancer Stem Cell Innovation Centre (SFI-CAST), Oslo University Hospital, Rikshospitalet, Oslo, NorwayUnit for Cell Signaling, Cancer Stem Cell Innovation Centre (SFI-CAST), Oslo University Hospital, Rikshospitalet, Oslo, NorwayTo whom correspondence should be addressed; Department of Chemistry, University of Oslo, Oslo, Norway; To whom correspondence should be addressedDepartment of Chemistry, University of Oslo, Oslo, NorwayIso-octyl chain-hydroxylated oxysterols were determined in attomoles per 10,000 cells concentrations in 10,000–80,000 cultured pancreatic adenocarcinoma cells, using a sensitive, highly automated nano-LC-ESI-MS-based method. Identified oxysterols included 24S hydroxycholesterol (24S-OHC), 25 hydroxycholesterol (25-OHC), and 27 hydroxycholesterol (27-OHC), while 20S hydroxycholesterol and 22S hydroxycholesterol were not detected. Lower mass limit of quantification was 23 fg (65 amol) for 25-OHC and 27-OHC (100 times lower than our previous method) and 54 fg (135 amol) for 24S-OHC, after derivatization into Girard T hydrazones and online sample cleanup using simplified and robust automatic filtration and filter back flushing solid phase extraction LC/MS/MS. The instrument configuration was easily installed using a commercial nano-LC/MS system. Recoveries in spiked sample were 96, 97, and 77% for 24S-OHC, 25-OHC, and 27-OHC, with within- and between-day repeatabilities of 1–21#x0025; and 2–20#x0025; relative SD, respectively. The study demonstrates the potential of nano-LC in lipidomics/sterolomics.http://www.sciencedirect.com/science/article/pii/S002222752035258524S hydroxycholesterol25 hydroxycholesterol27 hydroxycholesterolcholesterol 24-hydroxylasecholesterol 25-hydroxylasecholesterol 27-hydroxylase |
| spellingShingle | Hanne Roberg-Larsen Kaja Lund Tore Vehus Nina Solberg Caroline Vesterdal Dorna Misaghian Petter Angell Olsen Stefan Krauss Steven Ray Wilson Elsa Lundanes Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S] Journal of Lipid Research 24S hydroxycholesterol 25 hydroxycholesterol 27 hydroxycholesterol cholesterol 24-hydroxylase cholesterol 25-hydroxylase cholesterol 27-hydroxylase |
| title | Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S] |
| title_full | Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S] |
| title_fullStr | Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S] |
| title_full_unstemmed | Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S] |
| title_short | Highly automated nano-LC/MS-based approach for thousand cell-scale quantification of side chain-hydroxylated oxysterols[S] |
| title_sort | highly automated nano lc ms based approach for thousand cell scale quantification of side chain hydroxylated oxysterols s |
| topic | 24S hydroxycholesterol 25 hydroxycholesterol 27 hydroxycholesterol cholesterol 24-hydroxylase cholesterol 25-hydroxylase cholesterol 27-hydroxylase |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520352585 |
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