A high-performance liquid chromatography assay for measuring integrated biphenyl metabolism by intact cells: its use with rat liver and human liver and kidney.
A high-performance liquid chromatography assay for measuring integrated biphenyl metabolism by intact cells: its use with rat liver and human liver and kidney.
复制标题
一种用于测量完整细胞的综合联苯代谢的高效液相色谱测定法：其在大鼠肝脏以及人类肝脏和肾脏中的应用。
DOI：
10.1016/0003-2697(87)90151-5
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发表时间：
1987
影响因子：
2.9
通讯作者：
Santone,KS
中科院分区：
文献类型：
--
作者：
Powis,G;Moore,DJ;Wilke,TJ;Santone,KS
建立了一种快速、灵敏的高效液相色谱荧光检测方法，用于测定完整细胞的联苯代谢。该分析不需要有机溶剂萃取或酶消化对羟基联苯偶联物的测量。4-羟基联苯的检出下限为5 pmol。用联苯孵育的大鼠肝细胞主要形成4-羟基联苯硫酸盐、少量的4-羟基联苯葡萄糖醛酸和游离羟基联苯，以及少量的3-羟基联苯硫酸盐和3-羟基联苯葡萄糖醛酸。用联苯孵育的新鲜人肝片主要形成4-羟基联苯葡萄糖醛酸盐，还有一些游离的羟基联苯和少量的4-羟基联苯硫酸盐。4-羟基联苯葡萄糖醛酸盐在人肝脏中的形成有一个滞后时间，这个滞后时间不能通过无底物的肝脏预孵育来消除。用联苯形式4-羟基联苯葡萄糖醛酸盐和4-羟基联苯硫酸盐孵育人肾片，其孵育率低于用人肝脏孵育的十分之一。人体肾片不形成可检测到的游离羟基联苯。人体肝脏和肾脏形成羟基联苯代谢物的速率存在广泛的主体间变异性。
A rapid, sensitive high-performance liquid chromatography assay with fluorescence detection for measuring biphenyl metabolism by intact cells has been developed. The assay does not require organic solvent extraction or enzymatic digestion for the measurement of hydroxybiphenyl conjugates. The lower limit of detectability for 4-hydroxybiphenyl is 5 pmol injected. Rat hepatocytes incubated with biphenyl form predominantly 4-hydroxybiphenyl sulfate with lesser amounts of 4-hydroxybiphenyl glucuronide and free hydroxybiphenyls, and small amounts of 3-hydroxybiphenyl sulfate and 3-hydroxybiphenyl glucuronide. Slices of fresh human liver incubated with biphenyl form predominantly 4-hydroxybiphenyl glucuronide with some free hydroxybiphenyl and small amounts of 4-hydroxybiphenyl sulfate. 4-Hydroxybiphenyl glucuronide formation by human liver shows a lag time that is not abolished by preincubating the liver without substrate. Human kidney slices incubated with biphenyl form 4-hydroxybiphenyl glucuronide and 4-hydroxybiphenyl sulfate at rates less than one-tenth those seen with human liver. Human kidney slices do not form detectable free hydroxybiphenyl. There is wide intersubject variability in the rates of hydroxybiphenyl metabolite formation by human liver and kidney.