12 April 2005 Enantiomeric determination of D-, L-lactate in diabetic rat urine using a column-switching HPLC
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Proceedings Volume 5852, Third International Conference on Experimental Mechanics and Third Conference of the Asian Committee on Experimental Mechanics; (2005) https://doi.org/10.1117/12.621955
Event: Third International Conference on Experimental Mechanics and Third Conference of the Asian Committee on Experimental Mechanics, 2004, -, Singapore
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.
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Chien-Ming Chen, Chien-Ming Chen, Yih-Chiao Tsai, Yih-Chiao Tsai, Takeshi Fukushima, Takeshi Fukushima, Kazuhiro Imai, Kazuhiro Imai, Jen-Ai Lee, Jen-Ai Lee, } "Enantiomeric determination of D-, L-lactate in diabetic rat urine using a column-switching HPLC", Proc. SPIE 5852, Third International Conference on Experimental Mechanics and Third Conference of the Asian Committee on Experimental Mechanics, (12 April 2005); doi: 10.1117/12.621955; https://doi.org/10.1117/12.621955
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