Abstract
A method for the separation and quantitation of several important biological thiolamines is described. The procedure employs a C18 reversed-phase HPLC system to separate the dinitrophenyl derivatives of reduced and oxidized glutathione and cysteine and relies on an internal standard, Nε-methyllysine, to minimize experimental error. The method was validated in three matrices (water, HepG2 cell lysates, and mouse liver homogenates) using several criteria. The detector response was linear for the dinitrophenyl derivatives of glutathione, glutathione disulfide, cysteine, and cystine in the concentrations ranging from 10 to 50 nmol/ml. Inter- and intra-day variation, percent recovery in the biological matrices, and limits of detection and quantitation were determined. For the most accurate determination, it is essential that standard curves be produced daily and in the same matrix as that being analyzed.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1-12 |
| Number of pages | 12 |
| Journal | Journal of Chromatography B: Biomedical Sciences and Applications |
| Volume | 761 |
| Issue number | 1 |
| DOIs | |
| State | Published - Sep 15 2001 |
| Externally published | Yes |
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Keywords
- Cysteine
- Cystine
- Glutathione
- Glutathione disulfide
- Thiolamines
ASJC Scopus subject areas
- Chemistry(all)
Cite this
A new and versatile method for determination of thiolamines of biological importance. / Dominick, Pamela K.; Cassidy, Pamela; Roberts, Jeanette C.
In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 761, No. 1, 15.09.2001, p. 1-12.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A new and versatile method for determination of thiolamines of biological importance
AU - Dominick, Pamela K.
AU - Cassidy, Pamela
AU - Roberts, Jeanette C.
PY - 2001/9/15
Y1 - 2001/9/15
N2 - A method for the separation and quantitation of several important biological thiolamines is described. The procedure employs a C18 reversed-phase HPLC system to separate the dinitrophenyl derivatives of reduced and oxidized glutathione and cysteine and relies on an internal standard, Nε-methyllysine, to minimize experimental error. The method was validated in three matrices (water, HepG2 cell lysates, and mouse liver homogenates) using several criteria. The detector response was linear for the dinitrophenyl derivatives of glutathione, glutathione disulfide, cysteine, and cystine in the concentrations ranging from 10 to 50 nmol/ml. Inter- and intra-day variation, percent recovery in the biological matrices, and limits of detection and quantitation were determined. For the most accurate determination, it is essential that standard curves be produced daily and in the same matrix as that being analyzed.
AB - A method for the separation and quantitation of several important biological thiolamines is described. The procedure employs a C18 reversed-phase HPLC system to separate the dinitrophenyl derivatives of reduced and oxidized glutathione and cysteine and relies on an internal standard, Nε-methyllysine, to minimize experimental error. The method was validated in three matrices (water, HepG2 cell lysates, and mouse liver homogenates) using several criteria. The detector response was linear for the dinitrophenyl derivatives of glutathione, glutathione disulfide, cysteine, and cystine in the concentrations ranging from 10 to 50 nmol/ml. Inter- and intra-day variation, percent recovery in the biological matrices, and limits of detection and quantitation were determined. For the most accurate determination, it is essential that standard curves be produced daily and in the same matrix as that being analyzed.
KW - Cysteine
KW - Cystine
KW - Glutathione
KW - Glutathione disulfide
KW - Thiolamines
UR - http://www.scopus.com/inward/record.url?scp=0035885204&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035885204&partnerID=8YFLogxK
U2 - 10.1016/S0378-4347(01)00298-5
DO - 10.1016/S0378-4347(01)00298-5
M3 - Article
C2 - 11585123
AN - SCOPUS:0035885204
VL - 761
SP - 1
EP - 12
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
SN - 0378-4347
IS - 1
ER -