Capdevila A, Wagner C
Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232, USA.
Anal Biochem. 1998 Nov 15;264(2):180-4. doi: 10.1006/abio.1998.2839.
The low levels of S-adenosylmethionine (AdoMet) and S-adenosylhomocysteine (AdoHcy) in plasma can be measured by formation of the fluorescent isoindole derivatives of these compounds. The procedure involves an initial separation of AdoMet and AdoHcy in deproteinized plasma by HPLC on a C-8 column followed by derivatization with naphthalenedialdehyde and cyanide for 10 min at pH 9.0. The fluorescent derivatives of AdoMet and AdoHcy are then chromatographed by HPLC on a C-18 column monitored with a fluorescence monitor. The formation of the isoindole goes to 85% completion and the overall recovery of standards added to plasma is about 65%. Correction for recovery is made by addition of known quantities of AdoMet and AdoHcy to plasma. Normal values (+/- SE) for AdoMet were 102.7 nM +/- 9.9 and for AdoHcy were 22.7 +/- 3.1.
血浆中低水平的S-腺苷甲硫氨酸(AdoMet)和S-腺苷高半胱氨酸(AdoHcy)可通过这些化合物荧光异吲哚衍生物的形成来测定。该程序包括首先在C-8柱上通过HPLC对去蛋白血浆中的AdoMet和AdoHcy进行分离,然后用萘二醛和氰化物在pH 9.0下衍生化10分钟。然后,AdoMet和AdoHcy的荧光衍生物通过HPLC在C-18柱上进行色谱分析,并用荧光监测器进行监测。异吲哚的形成完成率达85%,添加到血浆中的标准品的总回收率约为65%。通过向血浆中添加已知量的AdoMet和AdoHcy来进行回收率校正。AdoMet的正常值(±SE)为102.7 nM±9.9,AdoHcy的正常值为22.7±3.1。