Drug-free RBC lysate and to identify areas of peak 5 corresponding nucleoside monophosphate, diphosphate and triphosphate. Stability t, after freezing and thawing investigated in samples from patients.鈻 RESULTS AND DISCUSSION LC MS / MS analysis. All thiopurine nucleotides were easily ionized into positive and negative ESI. The deprotonated molecular ion was observed in the negative ESI showed a Signal, t h twice Ago compared to the protonated molecular ion signal in ESI positive. The fragmentation patterns, however, were more favorable in the positive ionization mode, where all the nucleotides, the protonated nucleobase as the main product Ionenspezifit t. In contrast, fragmentation in negative ion mode observed extensively Agomelatine 5-HT receptor inhibitor Lich Including non-specific fragments to the fraction of phosphate. H3PO4 loss and fragmentation of the diphosphate were observed for all triphosphates. All diphosphates showed a loss of water and the loss of H3PO4, w Fragmented during the respective nucleobase monophosphates with a relatively low sensitivity. Overall, the sensitivity was at least 5 times h Forth in the positive ion mode. All product ion spectra are shown in Figure 2 SI p MRM Trnsfer Length and the corresponding optimized MS / MS parameters are summarized in Table S3. Separation of 5-nucleoside monophosphate, diphosphate, triphosphate and was by ion exchange HPLC on a weak anion exchanger-S Column with a gradient of pH 6 to 10.5 and decreasing a concentration of ammonium acetate from 10 mM to 1 mM obtained, as for other nucleotide previously.21 the two masses of the precursor shore ions and products of the 5 ionmasses thioguanosine methylthioinosine respective phosphates and 5 above differ only by a Da, fully to be separated from these connections to a St tion of the crest of the isotope to prevent. Typical chromatograms, the separation of nucleotides shown in Figure 2. Validation Test.
The method is in the range of 20 pmol / ml to 8.0 nmol / ml for TGMP, TGDP, TgTp, MeTGMP, MeTGDP, MeTGTP, TIMP and TITP linear and 100 pmol / ml to 40 nmol / ml for MeTIMP, MeTIDP and MeTITP. The correlation coefficients from 24 independent Ngigen groups were better than 0.9919 for all metabolites. At the lower detection limit of accuracy of better than 15%, and the accuracy varies between 20% for all analytes. The limit of detection was 5 pmol / ml for MeTGMP, MeTGDP, MeTGTP, 10 pmol / ml for TGMP, TGDP, TgTp, TIMP and TITP, and 25 pmol / ml for MeTIMP, MeTIDP and MeTITP with 3 S / N. Selectivity of t and Idarubicin 57852-57-0 specificity of t were divided into six Feeder Llig selected Hlten samples determined untreated RBC. Extracted ion chromatograms of RBC Wei S were compared with chromatograms of blank samples mixed with the lowest point calibration. For almost all analytes could not interfere with the expected retention time can not be detected. Only the extracted ion chromatogram of TITP shows a big s peak shortly before the expected retention time on TITP leading tosome background peaks with a liquid Surface of more than 30% of Fl Surface of the LLOQ TITP. Nevertheless, both the accuracy and variability T at LLOQ in the desired range with an accuracy between 7.2% and 9.8% and a reproducibility of 11.4%. Pr Precision and accuracy were studied at concentrations low, medium and high all thiopu.