Cov khoom

Txhawm rau kom muaj nuj nqis ntau yam metabolites ntawm 5-fluorouracil (5-FU) thiab ob lub endogenous monophosphate nucleotides, peb tau tsim ib txoj hauv kev los ntawm cov kua chromatography-tandem mass spectrometry (LC-MS/MS).Qhov kev ntsuam xyuas no tau tso cai rau kev txiav txim siab ntawm: (i) cov khoom tsim tawm ntawm 5-fluoro-2'-deoxyuridine-5'-monophosphate (5-FdUMP) los ntawm 5-FU lossis 5-fluoro-2'-deoxyuridine (5-FdUrd) , (ii) qhov cuam tshuam ntawm 5-FdUMP concentration ntawm intracellular 2′-deoxyuridine-5′-monophosphate (dUMP) / thymidine-5′-monophosphate (TMP) piv, thiab (iii) qhov tso tawm ntawm 5-FdUMP thiab 5-FU los ntawm tib neeg kab lis kev cai los ntawm ABC thauj.Raws li peb qhov kev sim, cov hlwb tau tsim nrog 5-FU lossis 5-FUrd.Tom qab ntawd, cellular proteins tau precipitated los ntawm methanol.Cov txheej txheem no tau muab siab rho tawm rov qab.Tsis tas li ntawd, txhawm rau ntsuas 5-FU thiab 5-FdUMP tso tawm los ntawm cov hlwb, peb tau ua kom muaj nuj nqis ntawm cov molecules hauv kab lis kev cai nruab nrab.Media tau raug txhaj ncaj qha (5-FU) lossis hauv qab mus rau theem kev rho tawm siv Oasis Wax extraction cartridge (5-FdUMP).Kev sib cais ntawm cov tshuaj ntsuam xyuas tau ua tiav ntawm dC18 Atlantis 3.5 μm, (100 mm × 2.1 mm id) kem nrog isocratic hom siv ammonium formate tsis / methanol / dej (5/5/90, v / v) raws li theem mobile.Lub sijhawm khiav tsis tshaj 6.2 min.Cov analytes tau ionized nyob rau hauv ib tug electrospray interface nyob rau hauv tsis zoo ion hom.Peb validated txoj kev tshaj li ntawm 2.5-150 ng mL −1 raws li cov tebchaw.Intra- thiab inter-assay variability yog qis dua 10% nyob rau xya hnub.Tag nrho cov tebchaw tau ruaj khov hauv cov hlwb lossis hauv kab lis kev cai nruab nrab thaum cov qauv tau khaws cia ntawm -20 ° C rau tsawg kawg ob lub lis piam, thiab tom qab peb lub voj voog khov-thaw.Tsis muaj cov nyhuv matrix tau pom nyob rau hauv ob qho tib si xov xwm.