@conference{
author = "Obradović, Darija and Nikolaevich Stavrianidi, Andrey and Alekseevich Shpigun, Oleg and Agbaba, Danica",
year = "2021",
abstract = "A possibility of predicting retention behaviour without a large number of preliminary
experiments is a significant segment of theoretical and experimental investigations. By using
an appropriate design to define experimental conditions (Design of Experiments, DoE), the
retention behaviour of compounds can be described as a function of the most important
parameters of chromatographic system. Further, the DoE methodology has shown successful
applicability to modeling retention under the environmentally friendly Supercritical Fluid
Chromatography (SFC) conditions (1). In the last decades, robustness of the SFC instruments
has been improved in order to minimize ecological risks, and routine application of the SFC
technique has been introduced into many pharmaceutical strategies (e.g., the Good
Manufacturing Practice, GMP) (2). Therefore, the aim of our study was to define the influence
of the main chromatographic factors on the retention behaviour of fourteen imidazoline and
serotonin receptor ligands under the SFC conditions. Using the Central Composite Design
(CCD) approach, retention characteristics (k) of the test compounds were examined on the
mixed-mode stationary phase, with the mixture of supercritical CO2 and methanolic
ammonium-formate (with an addition of 0.1% formic acid) used as mobile phase. We took into
the consideration the influence of the following factors: volume fraction of methanol in mobile
phase (20-30 %), ammonium-formate concentration (15-25 mM), and temperature deviation
(20-30 °C). The most important chromatographic factors were selected by the step-wise
multilinear regression (MLR), and their statistical significance was assessed using the ANOVA
analysis. Based on the results obtained, it was established that the retention characteristics
were significantly influenced by changing the methanol and ammonium-formate
concentrations in mobile phase (r > 0.90, p < 0.05), Figure 1. High degree of agreement (r >
0.98) was observed, when the theoretically predicted logk values for 35% and 15% volume
fraction of methanol in mobile phase were compared with the experimental ones. The
obtained results confirm successful applicability of the experimental design methodology in
order to perform a minimum number of experiments, as demonstrated upon an example of
modeling and predicting the retention behaviour of imidazoline and serotonin receptor
ligands under the SFC conditions., Mogućnost predviđanja retencije u odsustvu velikog broja prethodnih eksperimenata,
značajan je segment teorijskih i eksperimentalnih ispitivanja. Odgovarajućim dizajnom
eksperimentalnih uslova (Design of Experiments, DoE), a na osnovu minimalnog broja
eksperimenata, može se definisati retenciono ponašanje jedinjenja u funkciji najznačajnijih
parametara hromatografskog sistema. Primena DoE metodologije je takođe zabeležena i
prilikom modelovanja retencije u uslovima ekološki bezbedne, superkritične fluidne
hromatografije (Supercritical Fluid Chromatography, SFC) (1). Zbog minimalnih ekoloških
rizika, robusnost SFC instrumenata je poboljšana, a rutinska primena SFC tehnike je uvedena
i u regulatorne farmaceutske propise (npr. dobra proizvođačka praksa; Good Manufacturing
Practice-GMP) (2). Na osnovu toga, cilj ovog istraživanja obuhvatio je definisanje uticaja
najznačajnijih faktora hromatografskog sistema na retenciono ponašanje 14 odabranih
liganada imidazolinskih i serotoninskih receptora na mixed-mode stacionarnoj fazi u SFC
uslovima. Uticaj zapreminskog udela metanola (20-30 %), koncentracije amonijum-formijata
(15-25 mM) i temperature (20-30 °C) na vrednosti retencionih faktora (k) odabranih liganada
imidazolinskih i serotoninskih receptora je ispitana primenom centralnog kompozitnog
dizajna (Central Composite Design, CCD) na mixed-mode stacionarnoj fazi. Kao mobilna faza
korišćena je smeša superkritičnog CO2 i metanolnog rastvora amonijum-formijata uz dodatak
0,1% mravlje kiseline. Najznačajniji hromatografski faktori su izdvojeni step-wise postupkom
u višestrukoj linearnoj regresionoj analizi (Multiple Linear Regression, MLR), a njihova
statistička značajnost je procenjena primenom ANOVA testa. U konstruisanim retencionim
modelima, zapremina metanola i koncentracija pufera su pokazale najveći uticaj na retenciono
ponašanje testiranih jedinjenja (r > 0,90; p < 0,05), Slika 1. U koliko se teorijski predviđene
logk vrednosti uporede sa eksperimentalno dobijenim vrednostima na 35% i 15%
zapreminskim udelima metanola u mobilnoj fazi, uočava se da je prisutan visok stepen
slaganja (r > 0,98). Dobijeni rezultati potvrđuju uspešnu primenljivost metodologije
eksperimentalnog dizajna u cilju izvođenja minimalnog broja eksperimenata, prilikom
modelovanja i predikcije retencionog ponašanja liganda imidazolinskih i serotoninskih
receptora u SFC sistemima.",
publisher = "Savez farmaceutskih udruženja Srbije (SFUS)",
journal = "Arhiv za farmaciju",
title = "Modeling retention behaviour of imidazoline and serotonin receptor ligands under conditions of green SFC techique, Modelovanje retencionog ponašanja liganada imidazolinskih i serotoninskih receptora u uslovima ekološki bezbedne SFC tehnike",
volume = "71",
number = "5 suplement",
pages = "S126-S129",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4681"
}
