Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography

Yoshikazu Yoshida; Yasuharu Ohiwa; Masao Shimamura; Toru Izumi; Shozo Yoshida; Katsuyuki Takahashi; Shinichi Miyairi; Mizue Makimura; Akira Naganuma

doi:10.1248/jhs.49.527

Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography

Yoshikazu Yoshida, Yasuharu Ohiwa, Masao Shimamura, Toru Izumi, Shozo Yoshida, Katsuyuki Takahashi, Shinichi Miyairi, Mizue Makimura, Akira Naganuma

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

Abstract

For the accurate quantitation of glutathione (GSH), cysteine (Cys) and cysteinylglycine (CysGly) in human plasma by detection of fluorescence after derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F), a thiol-specific fluorogenic agent and high-performance liquid chromatography (HPLC), we attempted to identify optimum conditions for the derivatization and treatment of samples. The extent of derivatization with SBD-F of endogenous thiol compounds in human plasma was affected by several factors, such as the concentration of SBD-F and the reaction time. Acidification and deproteinization of plasma immediately after the separation of plasma from whole blood were essential and prevented the determination of inappropriately low values. The method developed in the present study allowed recovery that was closed to 100% for each compound. The mean values determined for 61 healthy individuals were as follows: Cys, 50.1 μM; CysGly, 12.2 μM; and GSH, 5.3 μM.

Original language	English
Pages (from-to)	527-530
Number of pages	4
Journal	Journal of Health Science
Volume	49
Issue number	6
DOIs	https://doi.org/10.1248/jhs.49.527
Publication status	Published - 2003 Dec
Externally published	Yes

Keywords

Cysteine
Cysteinylglycine
Glutathione
HPLC
Human plasma

ASJC Scopus subject areas

Toxicology
Health, Toxicology and Mutagenesis

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Yoshida, Y., Ohiwa, Y., Shimamura, M., Izumi, T., Yoshida, S., Takahashi, K., Miyairi, S., Makimura, M., & Naganuma, A. (2003). Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography. Journal of Health Science, 49(6), 527-530. https://doi.org/10.1248/jhs.49.527

Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography. / Yoshida, Yoshikazu; Ohiwa, Yasuharu; Shimamura, Masao; Izumi, Toru; Yoshida, Shozo; Takahashi, Katsuyuki; Miyairi, Shinichi; Makimura, Mizue; Naganuma, Akira.

In: Journal of Health Science, Vol. 49, No. 6, 12.2003, p. 527-530.

Research output: Contribution to journal › Article › peer-review

Yoshida, Y, Ohiwa, Y, Shimamura, M, Izumi, T, Yoshida, S, Takahashi, K, Miyairi, S, Makimura, M & Naganuma, A 2003, 'Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography', Journal of Health Science, vol. 49, no. 6, pp. 527-530. https://doi.org/10.1248/jhs.49.527

Yoshida Y, Ohiwa Y, Shimamura M, Izumi T, Yoshida S, Takahashi K et al. Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography. Journal of Health Science. 2003 Dec;49(6):527-530. https://doi.org/10.1248/jhs.49.527

Yoshida, Yoshikazu ; Ohiwa, Yasuharu ; Shimamura, Masao ; Izumi, Toru ; Yoshida, Shozo ; Takahashi, Katsuyuki ; Miyairi, Shinichi ; Makimura, Mizue ; Naganuma, Akira. / Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography. In: Journal of Health Science. 2003 ; Vol. 49, No. 6. pp. 527-530.

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AU - Yoshida, Yoshikazu

AU - Ohiwa, Yasuharu

AU - Shimamura, Masao

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AU - Yoshida, Shozo

AU - Takahashi, Katsuyuki

AU - Miyairi, Shinichi

AU - Makimura, Mizue

AU - Naganuma, Akira

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AB - For the accurate quantitation of glutathione (GSH), cysteine (Cys) and cysteinylglycine (CysGly) in human plasma by detection of fluorescence after derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F), a thiol-specific fluorogenic agent and high-performance liquid chromatography (HPLC), we attempted to identify optimum conditions for the derivatization and treatment of samples. The extent of derivatization with SBD-F of endogenous thiol compounds in human plasma was affected by several factors, such as the concentration of SBD-F and the reaction time. Acidification and deproteinization of plasma immediately after the separation of plasma from whole blood were essential and prevented the determination of inappropriately low values. The method developed in the present study allowed recovery that was closed to 100% for each compound. The mean values determined for 61 healthy individuals were as follows: Cys, 50.1 μM; CysGly, 12.2 μM; and GSH, 5.3 μM.

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Optimum conditions for derivatization of glutathione, cysteine and cysteinylglycine in human plasma with ammonium 7-fluorobenzo-2-oxa-1,3-diazole- 4-sulfonate for accurate quantitation by high-performance liquid chromatography

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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