Kayayyaki

Don ƙididdige ƙayyadaddun metabolites da yawa na 5-fluorouracil (5-FU) da monophosphate nucleotides guda biyu, mun ƙirƙiri wata hanya ta asali dangane da ruwa chromatography-tandem mass spectrometry (LC-MS/MS).Wannan tantancewar ta ba da izinin ƙayyade: (i) samar da intracellular na 5-fluoro-2'-deoxyuridine-5'-monophosphate (5-FdUMP) daga 5-FU ko 5-fluoro-2'-deoxyuridine (5-FdUrd) , (ii) tasiri na 5-FdUMP maida hankali a kan intracellular 2'-deoxyuridine-5'-monophosphate (dUMP) / thymidine-5'-monophosphate (TMP) rabo, da kuma (iii) da secretion iyakar 5-FdUMP da 5-FU daga ƙwayoyin al'adar ɗan adam ta masu jigilar ABC.A ƙarƙashin yanayin gwajin mu, an haɗa sel tare da 5-FU ko 5-FUrd.Sa'an nan, sunadaran sunadaran suna haɓaka ta hanyar methanol.Wannan hanya ta ba da babban farfadowar hakar.Bugu da kari, don auna 5-FU da 5-FdUMP ɓoye daga sel, mun aiwatar da ƙididdige waɗannan ƙwayoyin a cikin matsakaicin al'ada.An yi wa kafofin watsa labarai allura kai tsaye (5-FU) ko kuma an sami ingantaccen hakar lokaci ta amfani da harsashin hakar Oasis Wax (5-FdUMP).An yi rarrabuwar nazari akan dC18 Atlantis 3.5 μm, (100 mm × 2.1 mm id) shafi tare da yanayin isocratic ta amfani da buffer ammonium formate buffer/methanol/water (5/5/90, v/v) azaman lokacin wayar hannu.Lokacin gudu bai wuce 6.2 min ba.An sanya masu binciken ion a cikin mahallin lantarki a ƙarƙashin yanayin ion mara kyau.Mun inganta hanyar a kan kewayon 2.5-150 ng ml -1 bisa ga mahadi.Bambancin tsaka-tsaki da tsaka-tsaki ya kasance ƙasa da 10% sama da kwanaki bakwai.Duk mahadi sun tsaya tsayin daka a cikin sel ko a matsakaicin al'ada lokacin da aka adana samfurori a -20 ° C na akalla makonni biyu, kuma bayan daskare-narkewa uku.Ba a sami tasirin matrix a cikin kafofin watsa labarai biyu ba.