Phosphate-Modified Nucleotides: Conversions in Human Serum

References

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• HPLC was carried out using Nucleosil 100C18 column (4x150 mm) with a linear gradient of methanol (0–35% during 25 min, 35% for 5 min) in 0.05 M KH2PO4 (pH 6.8). The flow rate was 0.5 ml/min. The retention time was 23.6 min (HpAZT), 20.7 min (HpFLT), 16.8 min (HpThd), 21.4 min (FpAZT), 18.6 min (FpFLT), 18.6 min (FpThd), 14.2 min (pAZT), 12.5 min (pFLT), 8.6 min (pThd), 25.7 min (AZT), 22.9 min (FLT), 17.8 min (Thd).
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• Analysis was carried out on Merck Kieselgel 60F254 HPTLC plates in the following systems (v/v): dioxane:H2O:acetic acid, 90:10:0.2 (A); dioxane: NH4OH, 9:1 (B); 2-propanol:ethanol:NH4OH, 4:5:1 (C). Rf in system A, dA, 0.72; dI, 0.68; Hx, 0.57; in system B, dA, 0.35, dI, 0.27, Hx, 0.04; in system C, Hx, 0.44.
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