title: Indirect chiral separation of tryptophan enantiomers by high performance liquid chromatography with indirect chemiluminiscence detection.
abstract: In recent years, the study of chiral compounds in vivo has received much attention. In this study, a novel method based on high performance liquid chromatography (HPLC) coupled with chemiluminescence (CL) detection was developed for the separation of tryptophan (Trp) enantiomers. o-Phthalaldehyde and N-acetyl-l-cysteine were used as chiral derivatization reagents for Trp before it can be detected by HPLC-CL method. The separation was carried out on an ODS column using a mobile phase composed of methanol-0.01mol/L phosphate buffer (40/60, v/v). Under the optimum conditions, satisfactory results were obtained, including complete separation, good relative standard deviations and low detection limits. The applicability of the proposed method has been validated by determining Trp in biological samples. Linear responses (r>0.9990) were observed over the range of 2.5×10(-7) to 1.2×10(-5)g/mL of Trp enantiomers, with quantitation limit of 2.5×10(-7)g/mL. The assay method shows good specificity to Trp enantiomers, and thus it will have great potential application in clinical diagnosis. The mean extraction efficiency of Trp enantiomers in mice plasma samples were 98.48% and 97.40%, respectively. The mean relative standard deviation (RSD) of Trp enantiomers were <3%.
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, PR China. Electronic address: jie_0822@163.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhou
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, PR China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Chen
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, PR China.
authors.forename: Fang
authors.initials: F
authors.lastname: Sun
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, PR China.
authors.forename: Pei
authors.initials: P
authors.lastname: Luo
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, PR China.
authors.forename: Qiuzheng
authors.initials: Q
authors.lastname: Du
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, PR China.
authors.forename: Suzhen
authors.initials: S
authors.lastname: Zhao
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D016014:Linear Models; D008163:Luminescent Measurements / Q000379:methods*; D008165:Luminol; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012972:Sodium Hydroxide; D013237:Stereoisomerism; D014364:Tryptophan / Q000097:blood* / Q000737:chemistry*
chemical_list: D012972:Sodium Hydroxide; D008165:Luminol; D014364:Tryptophan
keywords: HPLC-CL; Indirect CL; Indirect chiral separation; Tryptophan