Multikriterijumski pristup optimizaciji hromatografskih metoda za farmaceutsku analizu perindopril t-butilamina

Abstract

Cilj ove doktorske disertacije bio je da se korišćenjem savremene metodologije koja uključuje optimizaciju metode multikriterijmskom pristupom i procenu robusnosti metode upotrebom eksperimentalnog dizajna, razviju metode reverzno–fazne i mikroemulzione tečne hromatografije za analizu perindopril t-butilamina i njegovih nečistoća (perindoprilat, Y31, Y32 i Y33). U prvom delu doktorske disertacije opisan je razvoj metode reverzno–fazne tečne hromatografije za analizu ispitivanih jedinjenja. Uzimajući u obzir savremeni koncept kojim se obezbeđuje razvoj metode odgovarajućih karakteristika, u fazi optimizacije metode primenjen je centralni kompozicioni dizajn sastavljen iz punog faktorskog dizajna 24, zvezda dizajna sa tačkama   0,5 sa 4 ponavljanja u centralnoj tački, kojim je ispitan uticaj faktora (sadržaj acetonitrila u mobilnoj fazi, pH mobilne faze, temperatura kolone i brzina protoka mobilne faze) na odabrane odgovore sistema. Na osnovu dobijenih matematičkih modela izvedeni su zaključci o uticaju faktora ali ne i o globalnom optimumu u ispitivanom eksperimentalnom području. U tom cilju, primenjen je multikriterijumski pristup tj. Deringerova funkcija poželjnih odgovora. Rezultati su omogućili definisanje globalnog optimuma a subjektivni parametri ispitani su primenom analize osetljivosti čime je potvrđen adekvatan izbor optimalnih uslova. Nakon toga, procenjena je robusnost postavljenog optimuma primenom pristupa koji uključuje primenu eksperimentalnog dizajna. Robusnost metode reverzno–fazne tečne hromatografije procenjena je primenom Plackett–Burman dizajna kojim je kroz 12 eksperimenta ispitano 7 pravih faktora a u cilju kompletiranja matrice eksperimenta dodata su 4 veštačka faktora (dummies). Analiza rezultata primenom statističkih i grafičkih metoda potvrdila je odgovarajuću robusnost optimuma a na osnovu rezultata su određeni i intervali neznačajnosti za značajne faktore, kao i parametri za procenu pogodnosti sistema razvijene metode reverzno–fazne tečne hromatografije za analizu perindopril t-butilamina i njegovih nečistoća (perindoprilat, Y31, Y32 i Y33). Na kraju, ispitani su i ostali parametri validacije i metoda je primenjena za analizu tableta sa perindopril t-butilaminom...

The goal of this doctorial dissertation was the development of reversed phase high performance liquid chromatography (RP–HPLC) as well as microemulsion liquid chromatography (MELC) methods for the analysis of perindopril t-butylamine and its impurities (perindoprilate, Y31, Y32 and Y33), by using contemporary approaches such as Multi-criteria decision-making method (MCDM) for methods optimisation and experimental design for methods robustness assessment. Development of RP–HPLC method for the analysis of the investigated compounds was desribed in the first part of the doctorial dissertation. Taking into account the modern concept that ensures development of the method with appropriate characteristics, Central Composite Design (CCD) with 24 full factorial design,   0,5 star design and 4 replicates in central point, was chosen for the method optimisation. Previously described CCD was applied for examination of factors (acetonitrile content in the mobile phase, pH of the mobile phase, column temperature and flow rate of the mobile phase) effects on chosen system responses. Based on the defined mathematical models, conclusions on factor effects were drawn, but not on the global optimum within the examined experimental domain, as well. In that aim, multicriteria approach, i.e. Derringer’s desirability function was applied. Results obtained enabled defining of global optimum. Since desirability function itself includes subjective parameters, Sensitivity Analysis was applied which confirmed adequate selection of optimal conditions. After that, robustness of the optimum set was examined by applying experimental design. Robustness of the RP–HPLC method was assessed by applying Plackett–Burman design for examining seven real factors by 12 experiments. Besides seven real factors, 4 dummy factors were included in order to complete the plan of the experiments. Analysis of the results obtained performed by statistical and graphical methods confirmed appropriate robustness of the optimum. In addition, for effects estimated to be significant, non–significant intervals were calculated as well as parameters for the assessment of system–suitability for the developed RP–HPLC method for the analysis of perindopril t-butylamine and its impurities (perindoprilate, Y31, Y32 and Y33). Finally, method validation parameters were examined, after which the method was applied for the analysis of perindopril t-butylamine tablets...