質(zhì)譜與紫外檢測器的異同

上一篇 / 下一篇 2007-08-22 01:02:16/ 個人分類：質(zhì)譜

轉(zhuǎn)自丁香園，作者：Cooks

你可以把質(zhì)譜理解為一個檢測器，就如常用的紫外檢測器一樣。

那我們想想，紫外檢測器能提供什么信號呢？紫外檢測器只能對那些在紫外區(qū)（一般為190-400 nm）有吸收的化合物有信號，也就是可以看到高出本底的色譜峰。這時候的信號僅能提供化合物相對量或者絕對量（需要標準物質(zhì)）的信息，假如單波長紫外檢測器改為DAD，那么這時候除了提供被分析物質(zhì)量的信息以外，還能提供化合物的定性信息（特征吸收峰）。但DAD的紫外圖譜作為定性的依據(jù)存在專署性差的問題（很明顯，一張紫外圖譜一般只有幾個峰，并且都是很寬的峰，表示的信息當然很有限了）。

那么質(zhì)譜作為檢測器能提供什么信息？

1、定量信息

2、定性信息

下面我們具體分析一下，從定量和定性兩方面質(zhì)譜到底優(yōu)于紫外的有哪些地方：

首先說定量，

(1)從可分析的化合物方面，質(zhì)譜大大超過UV，比如沒有紫外吸收或僅有末端吸收的化合物（皂甙、萜類、糖等），用質(zhì)譜分析具有很大的優(yōu)勢。一般只要化合物能夠被電離，就可以被分析。極性比較大的化合物可用ESI源，極性比較小的用APCI源，而對于那些極性非常小的化合物，比如多環(huán)芳烴類化合物，即便用APCI相應也很差，可以考慮用APPI。

(2)從靈敏度上看，一般質(zhì)譜比紫外高1-2個數(shù)量級，假如采用SRM模式，靈敏度會更高，我們測過的生物樣品最低可測到pg/mL(麥角甾酮類化合物)。

(3)從分析的速度和樣品的通量上看，假如你用HPLC-UV作定量的話，必須把要分析的化合物與其他所有干擾物質(zhì)分開，這個問題在分析體內(nèi)樣品的時候尤其嚴重。而用質(zhì)譜的話，大部分情況你根本不用考慮分離，就可以實現(xiàn)定量，我們以前做的許多生物利用度的項目的分析方法都是死時間出峰的（Rt~=2 min）。這樣的分析速度，我們每天可以測定大于200個樣本，這時單純的液相方法無法比擬的。

(4)當然還有其它更多的指標，這和不同廠家的儀器也有關，以后我們再詳細探討。

接下來再看看定性方面，我們以離子阱為例（定性是離子阱的強項）：

(1)提供整個結(jié)構的信息。相對于DAD僅提供官能團的信息，質(zhì)譜提供的信息豐富多了，尤其是多極質(zhì)譜。有時候我們僅通過質(zhì)譜就可以確定為之化合物的結(jié)構，給你一篇最新參考文獻（Rapid Commun. Mass Spectrom. 2006; 20: 2328-2342）

(2)輔助代謝產(chǎn)物鑒定，這方面也是質(zhì)譜應用很廣的一個領域。生物體內(nèi)的代謝產(chǎn)物，往往很微量，直接分離得到相當困難，假如沒有質(zhì)譜技術，現(xiàn)在代謝物的鑒定將會是非常困難的。一方面質(zhì)譜可以提供代謝物的碎片信息，輔助你推斷代謝物的結(jié)構，然后你可以通過微生物轉(zhuǎn)化或者合成的方法，得到代謝，從而最終確定代謝產(chǎn)物的結(jié)構；另一方面，也可以通過質(zhì)譜直接確定代謝產(chǎn)物的結(jié)構，比如一些二相代謝產(chǎn)物（葡萄糖醛酸結(jié)合物等）。

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液質(zhì)在中藥方面的應用主要（國外文獻）是集中在藥材的基礎研究上，比如使用多種質(zhì)譜技術快速鑒定中藥中的已知和未知化合物，而不需要像傳統(tǒng)的方法那樣進行植化分離，大大節(jié)省了成本和時間。
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