Nā huahana

No ka helu kikoʻī ʻana i kekahi mau metabolites o 5-fluorouracil (5-FU) a me ʻelua nucleotides monophosphate endogenous, ua kūkulu mākou i kahi ala kumu i hoʻokumu ʻia ma ka wai chromatography-tandem mass spectrometry (LC-MS/MS).Ua ʻae kēia hōʻike i ka hoʻoholo ʻana o: (i) ka hana intracellular o 5-fluoro-2′-deoxyuridine-5′-monophosphate (5-FdUMP) mai 5-FU a i ʻole 5-fluoro-2′-deoxyuridine (5-FdUrd) , (ii) ka hopena o ka 5-FdUMP hoʻonui i ka intracellular 2′-deoxyuridine-5′-monophosphate (dUMP) / thymidine-5′-monophosphate (TMP) lākiō, a (iii) ka nui huna o 5-FdUMP a 5-FU mai ke kanaka moʻomeheu pūnaewele e ABC transporters.Ma lalo o kā mākou kūlana hoʻokolohua, ua hoʻokomo ʻia nā cell me 5-FU a i ʻole 5-FUrd.A laila, ua hoʻoheheʻe ʻia nā protein cellular e ka methanol.Hāʻawi kēia kaʻina hana i ka hoʻihoʻi kiʻekiʻe.Eia kekahi, no ke ana ʻana i ka huna ʻana o 5-FU a me 5-FdUMP mai nā pūnaewele, ua hana mākou i ka helu ʻana o kēia mau mole i ka moʻomeheu moʻomeheu.Ua hoʻokomo pololei ʻia ka Media (5-FU) a i ʻole ma lalo o ka hoʻohana ʻana i ka pahu hoʻoheheʻe ʻo Oasis Wax (5-FdUMP).Hoʻokaʻawale ʻia nā analytes ma kahi kolamu dC18 Atlantis 3.5 μm, (100 mm × 2.1 mm id) me ke ʻano isocratic me ka hoʻohana ʻana i ka ammonium formate buffer / methanol / wai (5/5/90, v / v) e like me ka pae kelepona.ʻAʻole iʻoi aku ka manawa holo ma mua o 6.2 min.Ua hoʻokaʻawale ʻia nā analytes i kahi interface electrospray ma lalo o ke ʻano ion maikaʻi ʻole.Ua hōʻoia mākou i ke ʻano ma luna o kahi ākea o 2.5-150 ng mL −1 e like me nā pūhui.ʻOi aku ka liʻiliʻi o ka hoʻololi intra-a me inter-assay ma mua o 10% i nā lā ʻehiku.Paʻa nā pūhui a pau i loko o nā pūnaewele a i ʻole ka moʻomeheu i ka wā i mālama ʻia ai nā laʻana ma −20 °C no ka liʻiliʻi ʻelua pule, a ma hope o ʻekolu mau pōʻai hau.ʻAʻole i ʻike ʻia ka hopena matrix ma nā media ʻelua.