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準(zhǔn)博士金蟲 (正式寫手)
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[求助]
請(qǐng)教萃取內(nèi)標(biāo)法!
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According to the manuscript, the authors have analyzed the products of their bioconversion experiments after liquid-liquid extraction with ethyl acetate. However, there is no indication of the usage of an internal standard (such as progesterone, for example). According to my experience, this is a serious flaw. Typically, there is a significant variation in extraction efficiency from sample to sample, even if there is no change in the composition of the biotransformation media. This effect was neglected by the authors. 以上是審稿人提的意見。我先描述一下我的實(shí)驗(yàn):我做得是微生物轉(zhuǎn)化甾體化合物,轉(zhuǎn)化結(jié)束,采用乙酸乙酯萃取,然后旋光儀蒸干,接著用乙腈溶解,上液相,分析最終產(chǎn)物、底物和副產(chǎn)物。本身覺得應(yīng)該沒有問題,但因?yàn)榉蛛x提取沒系統(tǒng)研究過,審稿人的建議看不懂。請(qǐng)高人解釋。 |
銅蟲 (職業(yè)作家)
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大概了解審稿人的想法,他的意思是指即使你的biotransformation media都是一樣的話(assume轉(zhuǎn)化的結(jié)果都相同),但是每一批樣品間在經(jīng)過液液萃取后,其萃取效率都會(huì)有差別(there is a significant variation in extraction efficiency from sample to sample),造成你最后上液相后得到的產(chǎn)物與副產(chǎn)物會(huì)有一點(diǎn)差距,這是因?yàn)閷?shí)驗(yàn)條件變異造成的誤差,故應(yīng)在萃取前加入內(nèi)標(biāo)物,即使萃曲效率有些微變化,但分析物與內(nèi)標(biāo)物變化的比例是相同的,萃取造成的變異才會(huì)降到最低。 請(qǐng)參考看看。 |
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