Awọn ọja

Lati ṣe iwọn ni pato awọn metabolites pupọ ti 5-fluorouracil (5-FU) ati monophosphate nucleotides endogenous meji, a ṣe agbekalẹ ọna atilẹba ti o da lori chromatography olomi-tandem mass spectrometry (LC-MS/MS).Iwadii yii gba laaye ipinnu ti: (i) iṣelọpọ intracellular ti 5-fluoro-2′-deoxyuridine-5′-monophosphate (5-FdUMP) lati 5-FU tabi 5-fluoro-2′-deoxyuridine (5-FdUrd) , (ii) ikolu ti 5-FdUMP ifọkansi lori intracellular 2'-deoxyuridine-5'-monophosphate (dUMP)/thymidine-5'-monophosphate (TMP) ratio, ati (iii) iye yomijade ti 5-FdUMP ati 5-FU lati awọn sẹẹli gbin eniyan nipasẹ awọn gbigbe ABC.Labẹ awọn ipo idanwo wa, awọn sẹẹli ti wa pẹlu 5-FU tabi 5-FUrd.Lẹhinna, awọn ọlọjẹ cellular ti ṣaju nipasẹ methanol.Ilana yii pese imularada isediwon giga.Ni afikun, lati wiwọn 5-FU ati 5-FdUMP yomijade lati awọn sẹẹli, a ṣe iwọn awọn ohun elo wọnyi ni alabọde aṣa.Media ti wa ni itasi taara taara (5-FU) tabi labẹ ti lọ isediwon alakoso to lagbara nipa lilo katiriji isediwon Oasis Wax (5-FdUMP).Iyapa ti awọn atunnkanka ni a ṣe lori dC18 Atlantis 3.5 μm, (100 mm × 2.1 mm id) iwe pẹlu ipo isocratic nipa lilo ammonium formate buffer/methanol/water (5/5/90, v/v) gẹgẹbi apakan alagbeka.Akoko ṣiṣe ko kọja iṣẹju 6.2.Awọn atunnkanka naa jẹ ionized ni wiwo elekitiropiti labẹ ipo ion odi.A fọwọsi ọna naa lori iwọn 2.5-150 ng mL -1 ni ibamu si awọn agbo ogun.Iyipada inu-ati agbeyẹwo agbedemeji jẹ kekere ju 10% ju ọjọ meje lọ.Gbogbo awọn agbo ogun jẹ iduroṣinṣin ninu awọn sẹẹli tabi ni alabọde aṣa nigbati awọn ayẹwo ti wa ni ipamọ ni -20 °C fun o kere ju ọsẹ meji, ati lẹhin awọn iyipo didi-diẹ mẹta.Ko si ipa matrix ti a ṣe akiyesi ni awọn media mejeeji.