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

Fingerprint

lactates
urine
liquid chromatography
High performance liquid chromatography
rats
Rats
Derivatives
reagents
Wavelength
wavelengths
excitation

Keywords

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

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

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

Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC. / Chen, Chien Ming; Tsai, Yih Chiao; Fukushima, Takeshi; Imai, Kazuhiro; Lee, Jen Ai.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / C. Quan; F.S. Chau; A. Asundi; B.S. Wong; C.T. Lim. Vol. 5852 PART II 2005. p. 954-958 150.

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

Chen, CM, Tsai, YC, Fukushima, T, Imai, K & Lee, JA 2005, Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC. in C Quan, FS Chau, A Asundi, BS Wong & CT Lim (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 5852 PART II, 150, pp. 954-958, Third International Conference on Experimental Mechanics and Third Conference of the Asian Committee on Experimental Mechanics, Singapore, Singapore, 11/29/04. https://doi.org/10.1117/12.621955
Chen CM, Tsai YC, Fukushima T, Imai K, Lee JA. Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC. In Quan C, Chau FS, Asundi A, Wong BS, Lim CT, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 5852 PART II. 2005. p. 954-958. 150 https://doi.org/10.1117/12.621955
Chen, Chien Ming ; Tsai, Yih Chiao ; Fukushima, Takeshi ; Imai, Kazuhiro ; Lee, Jen Ai. / Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC. Proceedings of SPIE - The International Society for Optical Engineering. editor / C. Quan ; F.S. Chau ; A. Asundi ; B.S. Wong ; C.T. Lim. Vol. 5852 PART II 2005. pp. 954-958
@inproceedings{f99a3e4774d74d15a939f041314fbed5,
title = "Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC",
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.",
keywords = "Chiralpak AD-RH, D-lactate, Enantiomeric determination, NBD-PZ",
author = "Chen, {Chien Ming} and Tsai, {Yih Chiao} and Takeshi Fukushima and Kazuhiro Imai and Lee, {Jen Ai}",
year = "2005",
doi = "10.1117/12.621955",
language = "English",
volume = "5852 PART II",
pages = "954--958",
editor = "C. Quan and F.S. Chau and A. Asundi and B.S. Wong and C.T. Lim",
booktitle = "Proceedings of SPIE - The International Society for Optical Engineering",

}

TY - GEN

T1 - Enantiomeric determination of D-, L-Iactate in diabetic rat urine using a column-switching HPLC

AU - Chen, Chien Ming

AU - Tsai, Yih Chiao

AU - Fukushima, Takeshi

AU - Imai, Kazuhiro

AU - Lee, Jen Ai

PY - 2005

Y1 - 2005

N2 - 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.

AB - 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.

KW - Chiralpak AD-RH

KW - D-lactate

KW - Enantiomeric determination

KW - NBD-PZ

UR - http://www.scopus.com/inward/record.url?scp=28444479152&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=28444479152&partnerID=8YFLogxK

U2 - 10.1117/12.621955

DO - 10.1117/12.621955

M3 - Conference contribution

VL - 5852 PART II

SP - 954

EP - 958

BT - Proceedings of SPIE - The International Society for Optical Engineering

A2 - Quan, C.

A2 - Chau, F.S.

A2 - Asundi, A.

A2 - Wong, B.S.

A2 - Lim, C.T.

ER -