Abstract
Cholesterol is considered one of the most abundant sterols present in mammals, amphipathic in nature, and a key constituent of the cell membrane. Its unique chemical structure consisting of four linked hydrocarbon rings, with an aliphatic chain on one end and a hydroxyl group on the other, confers it the ability to form hydrogen bonds with other lipid classes, like phospholipids or sphingolipids head groups. However, due to its hydrophobic moiety, the fatty acid chain, free cholesterol (FC) it is difficult to be analyzed via electrospray ionization (ESI). The method presented in this chapter to identify and quantify free cholesterol and cholesteryl esters (CE) is based on the chemical derivatization of the sample, strategy devised to avoid the problematic of ESI. However, relevant mention should be made to an alternative separation protocol, which uses ultrahigh performance liquid chromatography and in-source collision-induced dissociation to achieve a simultaneous quantification of FC, CE, and triglycerides (TG).
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 53-59 |
| Number of pages | 7 |
| DOIs | |
| State | Published - Jan 1 2019 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1996 |
| ISSN (Print) | 1064-3745 |
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Keywords
- Cholesterol
- Cholesterol esters
- Electrospray ionization
- Lipidomics
- Mass spectrometry
- Orbitrap™
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Analyses of cholesterol and derivatives in ocular tissues using LC-MS/MS methods. / Piqueras, Maria; Theotoka, Despoina; Sarohia, Gurkaran S.; Bhattacharya, Sanjoy K.
Methods in Molecular Biology. Humana Press Inc., 2019. p. 53-59 (Methods in Molecular Biology; Vol. 1996).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Analyses of cholesterol and derivatives in ocular tissues using LC-MS/MS methods
AU - Piqueras, Maria
AU - Theotoka, Despoina
AU - Sarohia, Gurkaran S.
AU - Bhattacharya, Sanjoy K
PY - 2019/1/1
Y1 - 2019/1/1
N2 - Cholesterol is considered one of the most abundant sterols present in mammals, amphipathic in nature, and a key constituent of the cell membrane. Its unique chemical structure consisting of four linked hydrocarbon rings, with an aliphatic chain on one end and a hydroxyl group on the other, confers it the ability to form hydrogen bonds with other lipid classes, like phospholipids or sphingolipids head groups. However, due to its hydrophobic moiety, the fatty acid chain, free cholesterol (FC) it is difficult to be analyzed via electrospray ionization (ESI). The method presented in this chapter to identify and quantify free cholesterol and cholesteryl esters (CE) is based on the chemical derivatization of the sample, strategy devised to avoid the problematic of ESI. However, relevant mention should be made to an alternative separation protocol, which uses ultrahigh performance liquid chromatography and in-source collision-induced dissociation to achieve a simultaneous quantification of FC, CE, and triglycerides (TG).
AB - Cholesterol is considered one of the most abundant sterols present in mammals, amphipathic in nature, and a key constituent of the cell membrane. Its unique chemical structure consisting of four linked hydrocarbon rings, with an aliphatic chain on one end and a hydroxyl group on the other, confers it the ability to form hydrogen bonds with other lipid classes, like phospholipids or sphingolipids head groups. However, due to its hydrophobic moiety, the fatty acid chain, free cholesterol (FC) it is difficult to be analyzed via electrospray ionization (ESI). The method presented in this chapter to identify and quantify free cholesterol and cholesteryl esters (CE) is based on the chemical derivatization of the sample, strategy devised to avoid the problematic of ESI. However, relevant mention should be made to an alternative separation protocol, which uses ultrahigh performance liquid chromatography and in-source collision-induced dissociation to achieve a simultaneous quantification of FC, CE, and triglycerides (TG).
KW - Cholesterol
KW - Cholesterol esters
KW - Electrospray ionization
KW - Lipidomics
KW - Mass spectrometry
KW - Orbitrap™
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UR - http://www.scopus.com/inward/citedby.url?scp=85066242330&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-9488-5_6
DO - 10.1007/978-1-4939-9488-5_6
M3 - Chapter
C2 - 31127547
AN - SCOPUS:85066242330
T3 - Methods in Molecular Biology
SP - 53
EP - 59
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -