DETERMINATION OF GLUCOSINOLATES USING LIQUID CHROMATOGRAPHY WITH MASS SELECTIVE DETECTION

Namen diplomskega dela je bil določiti prisotnost sekundarnih metabolitov rastlin, iz skupine glukozinolatov, v vzorcih različnih križnic. Raziskave so potekale na tekočinskem kromatografu z masno selektivno detekcijo (LC/MS). Najprej smo iz pripravljenih čistih raztopin desetih standardov: glukonasturtin, glukonapin, glukoiberin, sinigrin, glukobrassicin, sinalbin, glukotropaeolin, epiprogoitrin, progoitrin in glukoraphenin določili pogoje, pri katerih bi lahko najučinkovitejše določali glukozinolate. Za dosego optimalnih pogojev smo spreminjali parametre, kot so: vrsta in sestava mobilne faze, kolona, pretok, napetost kolizijske celice, čas trajanja in območje meritev (m/z). Optimalni pogoji, ki smo jih določili, so: mobilna faza H2O z dodatkom metanojske kisline (HCOOH) in amonijevega formiata (CH5NO2), pretok skozi kolono 0,2 ml/min, napetost kolizijske celice 30 V in čas merjenja 30 minut. Ugotovili smo, da so glukozinolati prisotni v realnih vzorcih pri čemer se sestava spojin med vzorci spreminja, koncentracije glukozinolatov pa so praviloma nizke.The purpose of my diploma thesis was to determine the presence of secondary metabolites of plants, from the group of glucosinolates in a sample of cabbage and sacrum. The measurements were carried out on the liquid chromatography with mass selective detection (LC/MS), which allowed a quantitative and structural analysis. From prepared solutions of ten standardsgluconasturtin, gluconapin, glucoiberin, sinigrin, glucobrassicin, sinalbin, glucotropaeolin, epiprogoitrin, progoitrin and glucoraphenin, we first determined the conditions in which glucosinolates could be determined most effectively. To achieve appropriate conditions we changed the following parameters: mobile phase and its composition, the type of column and the flow through it, the voltage of the collision cell, duration and the area of measurements (m/z). Optimal conditions were determined as follows: H2O mobile phase containing formic acid (HCOOH) and ammonium formiate (CH5NO2), the flow through the column 0.2 ml / min, the collision cell voltage of 30 V and the measurement time of 30 minutes. We found that glucosinolates are present in real samples with the composition of the compounds varies between samples, the concentrations of glucosinolates are generally low