Zogulitsa

Kuti tidziwe bwino ma metabolites angapo a 5-fluorouracil (5-FU) ndi ma nucleotides awiri amtundu wa monophosphate, tinapanga njira yoyambirira yotengera madzi a chromatography-tandem mass spectrometry (LC-MS/MS).Kuyeza uku kunalola kutsimikiza kwa: (i) kupanga kwa intracellular ya 5-fluoro-2'-deoxyuridine-5'-monophosphate (5-FdUMP) kuchokera ku 5-FU kapena 5-fluoro-2'-deoxyuridine (5-FdUrd) + 5-FU kuchokera ku ma cell otukuka ndi anthu onyamula ABC.Pansi pa zoyeserera zathu, ma cell adalumikizidwa ndi 5-FU kapena 5-FUrd.Kenako, mapuloteni am'manja adapangidwa ndi methanol.Njira imeneyi anapereka mkulu m'zigawo kuchira.Kuphatikiza apo, kuyeza kutulutsa kwa 5-FU ndi 5-FdUMP kuchokera kumaselo, tidayesa kuchuluka kwa mamolekyuwa mu chikhalidwe chapakati.Media anali mwina mwachindunji jekeseni (5-FU) kapena anapita olimba gawo m'zigawo ntchito Oasis Sera m'zigawo katiriji (5-FdUMP).Kulekanitsa ma analytes kunachitika pa dC18 Atlantis 3.5 μm, (100 mm × 2.1 mm id) yokhala ndi isocratic mode pogwiritsa ntchito ammonium formate buffer/methanol/water (5/5/90, v/v) monga gawo la mafoni.Nthawi yothamanga sinapitirire 6.2 min.The analytes anali ionized mu mawonekedwe electrospray pansi negative ion mode.Tinatsimikizira njirayo pamtundu wa 2.5-150 ng mL -1 malinga ndi mankhwala.Kusiyana kwa intra- ndi inter-assay kunali kotsika kuposa 10% pamasiku asanu ndi awiri.Zosakaniza zonse zinali zokhazikika m'maselo kapena mu chikhalidwe cha chikhalidwe pamene zitsanzo zinkasungidwa pa -20 ° C kwa masabata osachepera awiri, ndipo pambuyo pa maulendo atatu oundana.Palibe zotsatira za matrix zomwe zidawonedwa muzofalitsa zonse ziwiri.