Alteration of biological samples in speciation analysis of metalloproteins.
For investigations of metalloproteins by speciation analysis, the integrity of the protein-metal complexes before and during separation is crucial. Knowledge about potential alterations of the samples is thus essential to avoid misinterpretations of the analytical results. Chromatographic element profiles of different cytosolic samples from animal tissues were measured repeatedly to estimate the sample stability. The dependence of the signals on the dwell time of the sample in an autosampling device at 4 degrees C for a period of 10 h was observed. Alterations in the element content of different metal-containing fractions were quantified by means of recovery values. Some metalloprotein fractions (e.g. approximately 27-kDa arsenic, approximately 27-kDa iron and different zinc fractions) were stable or only minor alterations were observed and for their investigation an autosampling device is therefore suitable. However, most of the other metalloprotein fractions, especially nickel-containing proteins, showed major alterations: these samples should therefore be analysed immediately after preparation or directly after thawing.
Department SF6, Hahn-Meitner-Institute Berlin, Glienicker Str. 100, 14109, Berlin, Germany. wolf@hmi.de
Christian
C
Wolf
Nadine
N
Wenda
Andrea
A
Richter
Antonios
A
Kyriakopoulos
D000818:Animals; D001331:Automation; D002845:Chromatography / Q000379:methods; D003600:Cytosol / Q000737:chemistry* / Q000378:metabolism; D007501:Iron / Q000032:analysis / Q000378:metabolism; D008667:Metalloproteins / Q000032:analysis* / Q000378:metabolism; D008970:Molecular Weight; D009532:Nickel / Q000032:analysis / Q000378:metabolism; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors
D008667:Metalloproteins; D009532:Nickel; D007501:Iron