Quantification of plasma S-adenosylmethionine and S-adenosylhomocysteine as their fluorescent 1,N(6)-etheno derivatives: an adaptation of previously described methodology.
A simplified reversed phase HPLC system for the detection of fluorescent 1,N(6)-etheno derivatives of SAM (S-adenosylmethionine) and S-adenosylhomocysteine (SAH) is described. The most important changes from the previously reported method are a shorter derivatization reaction time, the use of a solid-phase extraction resulting in an increase of the method's sensitivity, and the use of only one chromatographic system to separate SAM and SAH (in which the use of an ion-pairing reagent in the mobile phase is avoided). The linearity of the method was established, and the intra-assay coefficients of variation were 10.4 and 4.7% for SAM and SAH, respectively. Normal plasma values (n=8), evaluated with the present methodology, were, for SAM and SAH, respectively, 57+/-12 and 28+/-3 nM (mean+/-SD).
Castro
Rita
R
Department of Clinical Chemistry, Free University Hospital, Amsterdam, The Netherlands.
Struys
Eduard A
EA
Jansen
Erwin E W
EE
Blom
Henk J
HJ
de Almeida
Isabel Tavares
IT
Jakobs
Cornelis
C
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D015203:Reproducibility of Results; D012435:S-Adenosylhomocysteine / Q000097:blood* / Q000737:chemistry; D012436:S-Adenosylmethionine / Q000097:blood* / Q000737:chemistry; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine