TY - JOUR
T1 - Establishment of a two-dimensional chiral HPLC system for the simultaneous detection of lactate and 3-hydroxybutyrate enantiomers in human clinical samples
AU - Liu, Shu Ling
AU - Oyama, Tsubasa
AU - Miyoshi, Yurika
AU - Sheu, Shiow Yunn
AU - Mita, Masashi
AU - Ide, Tomomi
AU - Lindner, Wolfgang
AU - Hamase, Kenji
AU - Lee, Jen Ai
N1 - Funding Information:
This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan . The authors thank Shiseido Co., Ltd. (Tokyo, Japan) for technical support.
Publisher Copyright:
© 2015 Elsevier B.V.
PY - 2015/12/10
Y1 - 2015/12/10
N2 - A two-dimensional chiral high-performance liquid chromatography system was established for simultaneous detection of lactate (LA) and 3-hydroxybutyrate (3HB) enantiomers in human clinical samples. d-LA is increased upon kidney damage but 3HB protected against kidney injury. Therefore, determining the concentrations of d,. l-LA and d,. l-3HB simultaneously would be useful for evaluating pathological conditions. LA and 3HB were pre-column-derivatized with the fluorescent reagent 4-(. N-chloroformylmethyl-. N-methylamino)-7-nitro-2,1,3-benzoxadiazole (NBD-COCl) at 60. °C for 15. min and separated in the first dimension with a capillary monolithic octadecylsilane column. The mobile phase consisted of 13% acetonitrile and 0.05% tirfluoroacetic acid in water. Chiralpak QD-AX and KSAACSP-001S enantioselective columns were used in the second dimension to separate LA and 3HB enantiomers, respectively. Mobile phases were mixed solutions of methanol and acetonitrile containing formic acid. The separation factors were 1.14 and 1.08, respectively. The detection limit of LA and 3HB enantiomers was 10. fmol/injection. This method was applied to human clinical samples; intra- and inter-day relative standard deviations of LA and 3HB enantiomers were, respectively, 1.04-3.25% and 1.61-5.12% in plasma, 9.19-11.2% and 4.60-5.89% in urine, and 7.12-8.90% and 2.86-6.97% in saliva. This novel analytical method is a powerful tool for investigating variations in LA and 3HB enantiomers under disease conditions.
AB - A two-dimensional chiral high-performance liquid chromatography system was established for simultaneous detection of lactate (LA) and 3-hydroxybutyrate (3HB) enantiomers in human clinical samples. d-LA is increased upon kidney damage but 3HB protected against kidney injury. Therefore, determining the concentrations of d,. l-LA and d,. l-3HB simultaneously would be useful for evaluating pathological conditions. LA and 3HB were pre-column-derivatized with the fluorescent reagent 4-(. N-chloroformylmethyl-. N-methylamino)-7-nitro-2,1,3-benzoxadiazole (NBD-COCl) at 60. °C for 15. min and separated in the first dimension with a capillary monolithic octadecylsilane column. The mobile phase consisted of 13% acetonitrile and 0.05% tirfluoroacetic acid in water. Chiralpak QD-AX and KSAACSP-001S enantioselective columns were used in the second dimension to separate LA and 3HB enantiomers, respectively. Mobile phases were mixed solutions of methanol and acetonitrile containing formic acid. The separation factors were 1.14 and 1.08, respectively. The detection limit of LA and 3HB enantiomers was 10. fmol/injection. This method was applied to human clinical samples; intra- and inter-day relative standard deviations of LA and 3HB enantiomers were, respectively, 1.04-3.25% and 1.61-5.12% in plasma, 9.19-11.2% and 4.60-5.89% in urine, and 7.12-8.90% and 2.86-6.97% in saliva. This novel analytical method is a powerful tool for investigating variations in LA and 3HB enantiomers under disease conditions.
KW - 2D-HPLC
KW - 3-Hydroxybutyrate
KW - Chiral separation
KW - Lactate
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U2 - 10.1016/j.jpba.2015.05.036
DO - 10.1016/j.jpba.2015.05.036
M3 - Article
C2 - 26256241
AN - SCOPUS:84945437686
SN - 0731-7085
VL - 116
SP - 80
EP - 85
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
ER -
