Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC

Chien Ming Chen, Yih Chiao Tsai, Takeshi Fukushima, Kazuhiro Imai, Jen Ai Lee

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

Abstract

A highly sensitive method for the determination of D-lactate in rat urine was developed by using a high-performance liquid chromatography (HPLC) with an octadecylsilica (ODS) connected to a chiral column. At first, (D+L)-lactate in the urine were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1, 3-benzoxadiazole (NBD-PZ), and separated on the ODS column and determined fluorimetrically at 547 nm with 491 nm of excitation wavelength. During the separation step on the ODS, the peak fraction of (D+L)-lactate derivative was introduced directly to an amylose tris (3,5-dimethylphenylcarbamate) (Chiralpak AD-RH) chiral column by changing the flow of the eluent via 6-port valve. Then, D-lactate derivative was separated enantiomerically from L-lactate derivative, and the enantiomeric ratio was determined from the chromatogram. The accuracy values for the determination of D-lactate in 20 μL of rat urine were 96.93% -104.85%. The intra- and inter-day precision values were within 0.80% and 14.44%. The proposed method was applied to the urine of diabetic rats induced by intraperitoneal administration of streptozotocin, and the significant increases of D-lactate was observed in the diabetic rats as compared to the normal rats.

Original languageEnglish
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsC. Quan, F.S. Chau, A. Asundi, B.S. Wong, C.T. Lim
Pages954-958
Number of pages5
Volume5852 PART II
DOIs
Publication statusPublished - 2005
EventThird International Conference on Experimental Mechanics and Third Conference of the Asian Committee on Experimental Mechanics - Singapore, Singapore
Duration: Nov 29 2004Dec 1 2004

Other

OtherThird International Conference on Experimental Mechanics and Third Conference of the Asian Committee on Experimental Mechanics
CountrySingapore
CitySingapore
Period11/29/0412/1/04

Keywords

  • Chiralpak AD-RH
  • D-lactate
  • Enantiomeric determination
  • NBD-PZ

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

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  • Cite this

    Chen, C. M., Tsai, Y. C., Fukushima, T., Imai, K., & Lee, J. A. (2005). Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC. In C. Quan, F. S. Chau, A. Asundi, B. S. Wong, & C. T. Lim (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 5852 PART II, pp. 954-958). [150] https://doi.org/10.1117/12.621955