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xxfan

金虫 (著名写手)

[交流] 谷胱甘肽单乙酯合成(GSH-OEt)方法 已有1人参与

GSH-OEt购买的话,SIGMA是600多元25mg,1860元500mg,稍贵。
以下为其简便合成方法。
原来看到有人在求助,GSH-OEt哪里买?或者说怎么合成。
合成过程如下,和大家分享。
A freshly prepared solution of hydrogen
chloride (16 g, 0.45 mol) in 250 ml of
anhydrous ethanol was cooled to 20°C in
an ice bath, and 20 g (65 mmol) of glutathione
was added and dissolved by gentle
shaking. The mixture was allowed to stand
at 20°C for 2.5 h, during which time the
progress of the reaction was followed by
thin-layer chromatography (see Materials
and Methods). The solution was then
cooled to 0°C and 1.8 liter of ice-cold, dry
diethyl ether was added. The precipitate
which formed was allowed to settle for l-
3 h at 0°C. The ether was decanted and
the precipitate was washed twice with 500
ml of ice-cold ether. The product, after
drying in vacua over CaClz and KOH, was
obtained in 98% yield. The content of
glutathione monoethyl ester found on
amino acid analysis (see Materials and
Methods) varied from 90 to 98%; the other
components were glutathione and glutathione
diethyl ester. The preparations
obtained typically contained O-5% glutathione.

文献

Glutathione monoethyl ester: Preparation, uptake by tissues, and conversion to glutathione
Archives of biochemistry and biophysics
1985 Jun
239(2):538-48.
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xxfan

金虫 (著名写手)

引用回帖:
6楼: Originally posted by bbsoslolita at 2015-10-07 19:04:45
是用鼓吹氮气的方法来除氧的吧?嘿嘿...

先煮沸,再冷却,接着鼓吹氮气
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7楼2015-10-08 08:54:33
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bbsoslolita

银虫 (小有名气)


小木虫: 金币+0.5, 给个红包,谢谢回帖
为什么还原双硫键要用GSH-OEt啊?直接GSH不行吗?
2楼2015-10-01 11:46:18
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xxfan

金虫 (著名写手)

引用回帖:
2楼: Originally posted by bbsoslolita at 2015-10-01 11:46:18
为什么还原双硫键要用GSH-OEt啊?直接GSH不行吗?

GSH-OEt油水分配系数比较合适(脂溶性相对好些),GSH脂溶性太差,二者分别和同一株肿瘤细胞一起孵育时,GSH-OEt更容易透过细胞膜进入细胞质中(文献上是孵育2h,GSH-OEt即可100%进入细胞质,然后细胞质中酯酶参与降解GSH-OEt为GSH,达到需要的细胞质GSH浓度),而GSH不易进入细胞质。
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3楼2015-10-03 12:48:43
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bbsoslolita

银虫 (小有名气)


小木虫: 金币+0.5, 给个红包,谢谢回帖
引用回帖:
3楼: Originally posted by xxfan at 2015-10-03 12:48:43
GSH-OEt油水分配系数比较合适(脂溶性相对好些),GSH脂溶性太差,二者分别和同一株肿瘤细胞一起孵育时,GSH-OEt更容易透过细胞膜进入细胞质中(文献上是孵育2h,GSH-OEt即可100%进入细胞质,然后细胞质中酯酶参与 ...

嗯,原来是如此啊。那细胞外的实验,比如说载药胶束在细胞外的还原刺激性实验,就没有必要用GSH-OEt,直接GSH就可以了吧?
4楼2015-10-05 23:17:00
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