Toyo'oka T, Furukawa F, Suzuki T, Saito Y, Takahashi M, Hayashi Y, Uzu S, Imai K
National Institute of Hygienic Sciences, Tokyo, Japan.
Biomed Chromatogr. 1989 Jul;3(4):166-72. doi: 10.1002/bmc.1130030406.
Biological thiols and disulfides in rat and hamster tissues were simultaneously determined by HPLC-fluorescence detection using 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F) and ammonium 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate (SBD-F). The coefficients of variation (CV) of the method for reduced glutathione (GSH) and oxidized glutathione (GSSG) in liver and for cysteine (CySH) and cystine (CySSCy) in kidney were less than 3.1%. In 11 tissues of Wistar rats (liver, spleen, heart, lung, stomach, bladder, ovary, uterus, adrenal, kidney and pancreas), only CySH, CySSCy, GSH and/or GSSG were detected. Other thiols and disulfides were at extremely low levels in all samples. Both concentrations of CySH and CySSCy in the livers of old rats (111 weeks old, F344) were significantly higher than those of young rats (8 weeks old) (CySH, 0.246 +/- 0.099 vs 0.130 +/- 0.020 mumol/g; CySSCy, 0.051 +/- 0.027 vs 0.013 +/- 0.002 mumol/g). Administration of N-nitrosobis(2-oxopropyl)amine (BOP), a selective carcinogen of hamster pancreatic cancer, to Syrian golden hamsters (38 weeks old) resulted in the increase in the pancreas of GSH to a level 19 times as high and of GSSG to a level 14 times as high as those in untreated hamsters (GSH, 1.173 +/- 0.272 vs 0.062 +/- 0.017 mumol/g; GSSG, 0.155 +/- 0.063 vs 0.011 +/- 0.001 mumol/g).
采用4-(氨磺酰基)-7-氟-2,1,3-苯并恶二唑(ABD-F)和7-氟-2,1,3-苯并恶二唑-4-磺酸铵(SBD-F)，通过高效液相色谱-荧光检测法同时测定大鼠和仓鼠组织中的生物硫醇和二硫化物。该方法对肝脏中还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)以及肾脏中半胱氨酸(CySH)和胱氨酸(CySSCy)的变异系数(CV)小于3.1%。在Wistar大鼠的11种组织(肝脏、脾脏、心脏、肺、胃、膀胱、卵巢、子宫、肾上腺、肾脏和胰腺)中，仅检测到CySH、CySSCy、GSH和/或GSSG。所有样本中其他硫醇和二硫化物的含量极低。老年大鼠(111周龄，F344)肝脏中CySH和CySSCy的浓度均显著高于幼年大鼠(8周龄)(CySH，0.246±0.099对0.130±0.020μmol/g；CySSCy，0.051±0.027对0.013±0.002μmol/g)。给叙利亚金黄仓鼠(38周龄)注射仓鼠胰腺癌选择性致癌物N-亚硝基双(2-氧代丙基)胺(BOP)，导致胰腺中GSH水平升高至未处理仓鼠的19倍，GSSG水平升高至未处理仓鼠的14倍(GSH，1.173±0.272对0.062±0.017μmol/g；GSSG，0.155±0.063对0.011±0.001μmol/g)。
