TY - JOUR
T1 - High-performance liquid chromatographic technique for non-derivatized leucine purification
T2 - evidence for carbon isotope fractionation
AU - Baumann, Patricia Quinn
AU - Ebenstein, David B.
AU - O'Rourke, Bruce D.
AU - Nair, K. Sreekumaran
N1 - Funding Information:
This study was supported by Public Health Service Grant RO1 DK 41973-01 and General Clinical Research Centers Grant RR MO1 109. The authors would like to thank Tom Wheat of Waters Chromatography (Milford, MA, USA) for advice on non-derivatized amino acid separations.
PY - 1992/1/3
Y1 - 1992/1/3
N2 - A high-performance liquid chromatographic (HPLC) method, utilizing an ion-pairing agent with a reversed-phase column, isocratic elution and ultraviolet detection, was developed for the separation of non-derivatized leucine from a mixture of amino acids. Fractionation of leucine isotopes during HPLC separation was observed, creating potential problems for collection of leucine plasma or tissue protein hydrolyzates for subsequent isotopic determinations. It was demonstrated that [13C]leucine and [12C]leucine have different retention times (p<0.01). It is concluded from this study that partial collection of leucine eluted from an HPLC column will result in erroneous estimation of the isotopic enrichment of leucine.
AB - A high-performance liquid chromatographic (HPLC) method, utilizing an ion-pairing agent with a reversed-phase column, isocratic elution and ultraviolet detection, was developed for the separation of non-derivatized leucine from a mixture of amino acids. Fractionation of leucine isotopes during HPLC separation was observed, creating potential problems for collection of leucine plasma or tissue protein hydrolyzates for subsequent isotopic determinations. It was demonstrated that [13C]leucine and [12C]leucine have different retention times (p<0.01). It is concluded from this study that partial collection of leucine eluted from an HPLC column will result in erroneous estimation of the isotopic enrichment of leucine.
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U2 - 10.1016/0378-4347(92)80467-5
DO - 10.1016/0378-4347(92)80467-5
M3 - Article
C2 - 1564087
AN - SCOPUS:0026593025
SN - 0378-4347
VL - 573
SP - 11
EP - 16
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
IS - 1
ER -