TY - JOUR
T1 - Identification of L-3-hydroxybutyrate as an original ketone body in rat serum by column-switching high-performance liquid chromatography and fluorescence derivatization
AU - Tsai, Yih Chiao
AU - Liao, Tzu Hsin
AU - Lee, Jen Ai
N1 - Funding Information:
This work was financially supported by the National Science Council of the Republic of China (NSC 90-2320-B-038-029). The authors express their sincere appreciation to Professor K. Imai and Dr. T. Fukushima (Tokyo University, Tokyo, Japan) for their helpful suggestions.
PY - 2003/8/1
Y1 - 2003/8/1
N2 - L-3-Hydroxybutyrate (L-3HB), the enantiomer of D-3-hydroxybutyrate (D-3HB), has traditionally been regarded the "unnatural" ketone body in mammals, although there is suspicion that it is a more-favorable energy fuel for mammalian tissues than D-3HB. In this study, we demonstrated that L-3HB is an original substance in rat serum by applying fluorescence derivatization and a column-switching high-performance liquid chromatography system as the analysis technique. Racemic 3HB in rat serum derivatized using 4-nitro-7-piperazino-2,1,3-benzoxadiazole was first separated by an ODS column and was further confirmed by verifying the disappearance of the racemic 3HB peak after pretreating rat serum with D-3-hydroxybutyryl dehydrogenase. A switching valve was used to simultaneously introduce isolated racemic 3HB to the enantiomeric separation by two CHIRALCEL OD-RH columns connected in tandem. An L isomer was found to accompany the D isomer, which were quantified to be 3.98μM (3.61%) and 106.20μM (96.39%), respectively. Using the present analytical method, the dubious interpretation of the existence of L-3HB was clarified.
AB - L-3-Hydroxybutyrate (L-3HB), the enantiomer of D-3-hydroxybutyrate (D-3HB), has traditionally been regarded the "unnatural" ketone body in mammals, although there is suspicion that it is a more-favorable energy fuel for mammalian tissues than D-3HB. In this study, we demonstrated that L-3HB is an original substance in rat serum by applying fluorescence derivatization and a column-switching high-performance liquid chromatography system as the analysis technique. Racemic 3HB in rat serum derivatized using 4-nitro-7-piperazino-2,1,3-benzoxadiazole was first separated by an ODS column and was further confirmed by verifying the disappearance of the racemic 3HB peak after pretreating rat serum with D-3-hydroxybutyryl dehydrogenase. A switching valve was used to simultaneously introduce isolated racemic 3HB to the enantiomeric separation by two CHIRALCEL OD-RH columns connected in tandem. An L isomer was found to accompany the D isomer, which were quantified to be 3.98μM (3.61%) and 106.20μM (96.39%), respectively. Using the present analytical method, the dubious interpretation of the existence of L-3HB was clarified.
KW - Cellulose-based chiral column
KW - Column-switching HPLC
KW - Derivatization
KW - Enantiomeric separation
KW - L-3-Hydroxybutyrate
KW - NBD-PZ
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U2 - 10.1016/S0003-2697(03)00283-5
DO - 10.1016/S0003-2697(03)00283-5
M3 - Article
C2 - 12842104
AN - SCOPUS:0037677333
SN - 0003-2697
VL - 319
SP - 34
EP - 41
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -