Ngwaahịa

Iji kọwapụta ọtụtụ metabolites nke 5-fluorouracil (5-FU) na monophosphate nucleotides abụọ endogenous, anyị mepụtara usoro izizi dabere na chromatography nke tandem mass spectrometry (LC-MS/MS).Nyocha a nyere ohere ikpebi: (i) mmepụta intracellular nke 5-fluoro-2'-deoxyuridine-5'-monophosphate (5-FdUMP) sitere na 5-FU ma ọ bụ 5-fluoro-2'-deoxyuridine (5-FdUrd) , (ii) mmetụta nke 5-FdUMP ntinye uche na intracellular 2'-deoxyuridine-5'-monophosphate (dUMP)/thymidine-5'-monophosphate (TMP) ratio, na (iii) ihe nzuzo nke 5-FdUMP na 5-FU sitere na sel mmadụ mebere site na ndị njem ABC.N'okpuru ọnọdụ nnwale anyị, ejiri 5-FU ma ọ bụ 5-FUrd tinye mkpụrụ ndụ.Mgbe ahụ, methanol na-ebuli protein cellular.Usoro a nyere mgbake mmịpụta dị elu.Na mgbakwunye, iji tụọ 5-FU na 5-FdUMP nzuzo sitere na mkpụrụ ndụ, anyị mere ọnụọgụ nke ụmụ irighiri ihe ndị a na usoro ọdịnala.A na-agbanye mgbasa ozi ozugbo (5-FU) ma ọ bụ n'okpuru wepụta mmịpụta siri ike site na iji Oasis Wax extract cartridge (5-FdUMP).Emere nkewa nke nyocha na dC18 Atlantis 3.5 μm, (100 mm × 2.1 mm id) kọlụm nwere ọnọdụ isocratic na-eji ammonium formate buffer/methanol/water (5/5/90, v/v) dị ka usoro mkpanaka.Oge ịgba ọsọ agafeghị 6.2 min.A na-eme ka ndị nyocha ahụ ionized na ntanetị electrospray n'okpuru ọnọdụ ion na-adịghị mma.Anyị kwadoro usoro ahụ n'ogo nke 2.5-150 ng ml -1 dịka ogige ndị ahụ.Mgbanwe intra- na n'etiti nyocha dị ala karịa 10% n'ime ụbọchị asaa.Ogige niile kwụsiri ike na sel ma ọ bụ na usoro ọdịnala mgbe echekwara nlele na -20 Celsius C ma ọ dịkarịa ala izu abụọ, na mgbe usoro nchara atọ gachara.Enweghị mmetụta matrix ahụ na mgbasa ozi abụọ ahụ.