Abstract
In vivo determinations of amino acids are important for improving our understanding of physiological states of biological tissue function and dysfunction. However, the chemically complex matrix of different biological fluids complicates the assay of this important class of molecules. We introduce a method for characterizing the amino acid composition of submicroliter volumes of vitreous humor perfusates. Low-flow push-pull perfusion sampling is compatible with collecting small volume samples in a complicated matrix that are potentially difficult to separate. An efficient, sensitive, and rapid analysis of amino acids from in vivo perfusates of the vitreous is presented with 3-(4-carboxybenzoyl)-2-quinoline-carboxaldehyde (CBQCA) derivatitation and capillary electrophoresis (CE) separation with laser-induced fluorescence detection (LIF). Derivatization with CBQCA for up to 2 h provided high sensitivity and low detection limits at the nM level. Seventeen amino acids including D-serine(D-Ser) and D-aspartate (D-Asp) were resolved in less than 10 min. Importantly, D-Ser is separated from its enantiomeric pair. Characterization of vitreal amino acids with this assay technique will be useful for understanding ocular diseases and physiological mechanisms in vision.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2978-2984 |
| Number of pages | 7 |
| Journal | Electrophoresis |
| Volume | 25 |
| Issue number | 17 |
| DOIs | |
| State | Published - Sep 2004 |
| Externally published | Yes |
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Keywords
- Amino acids
- Chiral separation
- Flourescence
- Small-volume samples
- Vitreous samples
ASJC Scopus subject areas
- Clinical Biochemistry
Cite this
Determination of amino acids in rat vitreous perfusates by capillary electrophoresis. / Thongkhao-On, Kongthong; Kottegoda, Sumith; Pulido, Jose S; Shippy, Scott A.
In: Electrophoresis, Vol. 25, No. 17, 09.2004, p. 2978-2984.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Determination of amino acids in rat vitreous perfusates by capillary electrophoresis
AU - Thongkhao-On, Kongthong
AU - Kottegoda, Sumith
AU - Pulido, Jose S
AU - Shippy, Scott A.
PY - 2004/9
Y1 - 2004/9
N2 - In vivo determinations of amino acids are important for improving our understanding of physiological states of biological tissue function and dysfunction. However, the chemically complex matrix of different biological fluids complicates the assay of this important class of molecules. We introduce a method for characterizing the amino acid composition of submicroliter volumes of vitreous humor perfusates. Low-flow push-pull perfusion sampling is compatible with collecting small volume samples in a complicated matrix that are potentially difficult to separate. An efficient, sensitive, and rapid analysis of amino acids from in vivo perfusates of the vitreous is presented with 3-(4-carboxybenzoyl)-2-quinoline-carboxaldehyde (CBQCA) derivatitation and capillary electrophoresis (CE) separation with laser-induced fluorescence detection (LIF). Derivatization with CBQCA for up to 2 h provided high sensitivity and low detection limits at the nM level. Seventeen amino acids including D-serine(D-Ser) and D-aspartate (D-Asp) were resolved in less than 10 min. Importantly, D-Ser is separated from its enantiomeric pair. Characterization of vitreal amino acids with this assay technique will be useful for understanding ocular diseases and physiological mechanisms in vision.
AB - In vivo determinations of amino acids are important for improving our understanding of physiological states of biological tissue function and dysfunction. However, the chemically complex matrix of different biological fluids complicates the assay of this important class of molecules. We introduce a method for characterizing the amino acid composition of submicroliter volumes of vitreous humor perfusates. Low-flow push-pull perfusion sampling is compatible with collecting small volume samples in a complicated matrix that are potentially difficult to separate. An efficient, sensitive, and rapid analysis of amino acids from in vivo perfusates of the vitreous is presented with 3-(4-carboxybenzoyl)-2-quinoline-carboxaldehyde (CBQCA) derivatitation and capillary electrophoresis (CE) separation with laser-induced fluorescence detection (LIF). Derivatization with CBQCA for up to 2 h provided high sensitivity and low detection limits at the nM level. Seventeen amino acids including D-serine(D-Ser) and D-aspartate (D-Asp) were resolved in less than 10 min. Importantly, D-Ser is separated from its enantiomeric pair. Characterization of vitreal amino acids with this assay technique will be useful for understanding ocular diseases and physiological mechanisms in vision.
KW - Amino acids
KW - Chiral separation
KW - Flourescence
KW - Small-volume samples
KW - Vitreous samples
UR - http://www.scopus.com/inward/record.url?scp=4644275196&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=4644275196&partnerID=8YFLogxK
U2 - 10.1002/elps.200405941
DO - 10.1002/elps.200405941
M3 - Article
C2 - 15349938
AN - SCOPUS:4644275196
VL - 25
SP - 2978
EP - 2984
JO - Electrophoresis
JF - Electrophoresis
SN - 0173-0835
IS - 17
ER -