title: Establishment of a two-dimensional chiral HPLC system for the simultaneous detection of lactate and 3-hydroxybutyrate enantiomers in human clinical samples.
abstract: 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.
authors.affiliation: School of Pharmacy, College of Pharmacy, Taipei Medical University, No. 250 Wuxing St., Taipei 11031, Taiwan.
authors.forename: Shu-Ling
authors.initials: SL
authors.lastname: Liu
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Tsubasa
authors.initials: T
authors.lastname: Oyama
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Yurika
authors.initials: Y
authors.lastname: Miyoshi
authors.affiliation: School of Pharmacy, College of Pharmacy, Taipei Medical University, No. 250 Wuxing St., Taipei 11031, Taiwan.
authors.forename: Shiow-Yunn
authors.initials: SY
authors.lastname: Sheu
authors.affiliation: Shiseido Co., Ltd., 1-6-2 Higashi-shimbashi, Minato-ku, Tokyo 105-8310, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Mita
authors.affiliation: Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Ide
authors.affiliation: Institute of Analytical Chemistry, University of Vienna, Waehringerstrasse 38, A-1090 Vienna, Austria.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lindner
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. Electronic address: hamase@phar.kyushu-u.ac.jp.
authors.forename: Kenji
authors.initials: K
authors.lastname: Hamase
authors.affiliation: School of Pharmacy, College of Pharmacy, Taipei Medical University, No. 250 Wuxing St., Taipei 11031, Taiwan. Electronic address: jenai@tmu.edu.tw.
authors.forename: Jen-Ai
authors.initials: JA
authors.lastname: Lee
mesh_terms: D020155:3-Hydroxybutyric Acid / Q000032:analysis* / Q000378:metabolism*; D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods; D006801:Humans; D019344:Lactic Acid / Q000032:analysis* / Q000378:metabolism*; D008297:Male; D013237:Stereoisomerism; D055815:Young Adult
chemical_list: D019344:Lactic Acid; D020155:3-Hydroxybutyric Acid
keywords: 2D-HPLC; 3-Hydroxybutyrate; Chiral separation; Lactate