Determination of time-dependent accumulation of d-lactate in the streptozotocin-induced diabetic rat kidney by column-switching HPLC with fluorescence detection

Mei Hsiang Lin, Hsiang Yin Chen, Tzu Hsin Liao, Tzu Chuan Huang, Chien Ming Chen, Jen Ai Lee

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Abstract

For better understanding the complete metabolism and the physiological role of d-lactate, the concentrations of d-lactate in the serum, liver and kidney of normal and diabetic rats were determined by our established column-switching HPLC method with pre-column fluorescence derivatization. Eight-week-old male Sprague-Dawley rats were administered with streptozotocin (STZ) (80. mg/kg) or citrate buffer intraperitoneally. The tissues were then removed and homogenized after 4, 8, 12 and 16. weeks of drug administration, respectively. The homogenates were centrifuged at 1200 × g for 10. min, then the supernatants were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on an ODS column followed by a Chiralpak AD-RH chiral column for enantioseparation. The results showed that the d-lactate content elevated in all the 3 examined tissues under diabetic stages. In addition, d-lactate concentrations in rat kidney were accumulated significantly and time-dependently in diabetic groups after receiving STZ for 4, 8, 12 and 16. weeks (2.99, 13.11, 18.19, 23.23 vs. 0.79 μmol/mg protein as control group). Moreover, the kidney of induced 12-week diabetic rat renal showed some histological changes of progressive diabetic nephropathy. The results suggest that d-lactate may be used as a marker of diabetic nephropathy.

Original languageEnglish
Pages (from-to)3214-3219
Number of pages6
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume879
Issue number29
DOIs
Publication statusPublished - Nov 1 2011

Fingerprint

Streptozocin
Rats
Lactic Acid
Fluorescence
High Pressure Liquid Chromatography
Kidney
Diabetic Nephropathies
Tissue
Metabolism
Citric Acid
Liver
Sprague Dawley Rats
Buffers
Control Groups
Serum
Pharmaceutical Preparations
Proteins
4-nitro-7-piperazino-2,1,3-benzoxadiazole

Keywords

  • D-Lactate
  • Diabetic rat
  • L-Lactate
  • Methylglyoxal
  • Nephropathy
  • Streptozotocin

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Cell Biology
  • Clinical Biochemistry

Cite this

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title = "Determination of time-dependent accumulation of d-lactate in the streptozotocin-induced diabetic rat kidney by column-switching HPLC with fluorescence detection",
abstract = "For better understanding the complete metabolism and the physiological role of d-lactate, the concentrations of d-lactate in the serum, liver and kidney of normal and diabetic rats were determined by our established column-switching HPLC method with pre-column fluorescence derivatization. Eight-week-old male Sprague-Dawley rats were administered with streptozotocin (STZ) (80. mg/kg) or citrate buffer intraperitoneally. The tissues were then removed and homogenized after 4, 8, 12 and 16. weeks of drug administration, respectively. The homogenates were centrifuged at 1200 × g for 10. min, then the supernatants were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on an ODS column followed by a Chiralpak AD-RH chiral column for enantioseparation. The results showed that the d-lactate content elevated in all the 3 examined tissues under diabetic stages. In addition, d-lactate concentrations in rat kidney were accumulated significantly and time-dependently in diabetic groups after receiving STZ for 4, 8, 12 and 16. weeks (2.99, 13.11, 18.19, 23.23 vs. 0.79 μmol/mg protein as control group). Moreover, the kidney of induced 12-week diabetic rat renal showed some histological changes of progressive diabetic nephropathy. The results suggest that d-lactate may be used as a marker of diabetic nephropathy.",
keywords = "D-Lactate, Diabetic rat, L-Lactate, Methylglyoxal, Nephropathy, Streptozotocin",
author = "Lin, {Mei Hsiang} and Chen, {Hsiang Yin} and Liao, {Tzu Hsin} and Huang, {Tzu Chuan} and Chen, {Chien Ming} and Lee, {Jen Ai}",
year = "2011",
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T1 - Determination of time-dependent accumulation of d-lactate in the streptozotocin-induced diabetic rat kidney by column-switching HPLC with fluorescence detection

AU - Lin, Mei Hsiang

AU - Chen, Hsiang Yin

AU - Liao, Tzu Hsin

AU - Huang, Tzu Chuan

AU - Chen, Chien Ming

AU - Lee, Jen Ai

PY - 2011/11/1

Y1 - 2011/11/1

N2 - For better understanding the complete metabolism and the physiological role of d-lactate, the concentrations of d-lactate in the serum, liver and kidney of normal and diabetic rats were determined by our established column-switching HPLC method with pre-column fluorescence derivatization. Eight-week-old male Sprague-Dawley rats were administered with streptozotocin (STZ) (80. mg/kg) or citrate buffer intraperitoneally. The tissues were then removed and homogenized after 4, 8, 12 and 16. weeks of drug administration, respectively. The homogenates were centrifuged at 1200 × g for 10. min, then the supernatants were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on an ODS column followed by a Chiralpak AD-RH chiral column for enantioseparation. The results showed that the d-lactate content elevated in all the 3 examined tissues under diabetic stages. In addition, d-lactate concentrations in rat kidney were accumulated significantly and time-dependently in diabetic groups after receiving STZ for 4, 8, 12 and 16. weeks (2.99, 13.11, 18.19, 23.23 vs. 0.79 μmol/mg protein as control group). Moreover, the kidney of induced 12-week diabetic rat renal showed some histological changes of progressive diabetic nephropathy. The results suggest that d-lactate may be used as a marker of diabetic nephropathy.

AB - For better understanding the complete metabolism and the physiological role of d-lactate, the concentrations of d-lactate in the serum, liver and kidney of normal and diabetic rats were determined by our established column-switching HPLC method with pre-column fluorescence derivatization. Eight-week-old male Sprague-Dawley rats were administered with streptozotocin (STZ) (80. mg/kg) or citrate buffer intraperitoneally. The tissues were then removed and homogenized after 4, 8, 12 and 16. weeks of drug administration, respectively. The homogenates were centrifuged at 1200 × g for 10. min, then the supernatants were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on an ODS column followed by a Chiralpak AD-RH chiral column for enantioseparation. The results showed that the d-lactate content elevated in all the 3 examined tissues under diabetic stages. In addition, d-lactate concentrations in rat kidney were accumulated significantly and time-dependently in diabetic groups after receiving STZ for 4, 8, 12 and 16. weeks (2.99, 13.11, 18.19, 23.23 vs. 0.79 μmol/mg protein as control group). Moreover, the kidney of induced 12-week diabetic rat renal showed some histological changes of progressive diabetic nephropathy. The results suggest that d-lactate may be used as a marker of diabetic nephropathy.

KW - D-Lactate

KW - Diabetic rat

KW - L-Lactate

KW - Methylglyoxal

KW - Nephropathy

KW - Streptozotocin

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