Farmakokinetički parametri fitoestrogena
| Autor: | Lasić, Kornelija |
|---|---|
| Přispěvatelé: | Nigović, Biljana |
| Jazyk: | chorvatština |
| Rok vydání: | 2020 |
| Předmět: |
phytoestrogens
fitoestrogeni izoflavoni lipofilnost biomimetička kromatografija Caco-2 permeabilnost kromatografija visoke djelotvornosti kromatografija ultravisoke djelotvornosti kvaliteta ugrađena u dizajn dvodimenzionalna tekućinska kromatografija masena spektrometrija fitoestrogeni kromatografija ultravisoke djelotvornosti kromatografija visoke djelotvornosti Pharmacology. Therapeutics. Toxicology biomimetička kromatografija masena spektrometrija dvodimenzionalna tekućinska kromatografija udc:615(043.3) Caco-2 permeabilnost BIOMEDICINE AND HEALTHCARE. Pharmacy. Pharmacy lipofilnost ultra-high performance liquid chromatography izoflavoni Farmakologija. Terapeutika. Toksikologija lipophilicity two dimensional liquid chromatography Caco-2 permeability isoflavones biomimetic chromatography BIOMEDICINA I ZDRAVSTVO. Farmacija. Farmacija mass spectrometry |
| Popis: | Primjena fitoestrogena se kontinuirano povećava zbog njihove biološke aktivnosti i potencijalno korisnih učinaka. Na tržištu se nalazi sve veći broj pripravaka koji sadrže različite fitoestrogene namijenjene ženama u postmenopauzi kao posebno rizičnoj skupini. Osim što se razlikuju kvantitativno s obzirom na ukupni sadržaj fitoestrogena, dodatno se razlikuju i kvalitativno s obzirom na zastupljenost pojedinih fitoestrogena. Cilj je ovog rada odrediti farmakokinetičke parametre skupini 15 strukturno sličnih fitoestrogena (genistein, biokanin A, daidzein, formononetin, genistin, sisotrin, daidzin, ononin, prunetin, 3,4,7-trihidroksi izoflavon, 4,6,7-trihidroksi izoflavon, 4,6,7-trimetoksi izoflavon, genistein 4,7-dimetil eter, kumestrol, kumestrol dimetil eter) u svrhu boljeg razumijevanja njihove bioraspoloživosti i učinaka na organizam. Zatim, na temelju kromatografski određene lipofilnosti razviti novu, brzu, selektivnu i osjetljivu analitičku metodu za određivanje 8 najznačajnijih fitoestrogena iz skupine izoflavona u obliku aglikona i njihovih glikozida. Određeni su parametri koji opisuju lipofilnost skupine strukturno sličnih fitoestrogena primjenom tekućinske kromatografije ultravisoke djelotvornosti obrnutih faza (RP-UHPLC). Ispitane su njihove interakcije s fosfolipidima staničnih membrana primjenom tekućinske kromatografije visoke djelotvornosti i kolona s vezanim umjetnim membranama. Određeni su udjeli vezanja fitoestrogena za najzastupljenije proteine plazme primjenom biomimetičke kromatografije. Permeabilnost fitoestrogena i način prijenosa kroz sluznicu gastrointestinalnog trakta određeni su primjenom modela Caco-2 stanica. Dobiveni rezultati diferenciraju fitoestrogene sukladno njihovoj strukturi, gdje su aglikoni pokazali značajno višu lipofilnost, jaču interakciju s fosfolipidima, viši udio vezanja za alfa-1-kiseli glikoprotein i višu permeabilnost u usporedbi s glikozidima. Međutim, udio vezanja za humani serumski albumin bio je visok za sve ispitivane fitoestrogene. Eksperimentalno određeni farmakokinetički parametri uspoređeni su s izračunatim različitim računalnim programima. Razvijena je nova RP-UHPLC metoda primjenom QbD pristupa za istovremenu kvntifikaciju najznačajnijih fitoestrogena iz skupine izoflavona u obliku glikozida (genistin, sisotrin, daidzin, ononin) i aglikona (genistein, biokanin A, daidzein, formononetin) bez prethodno provedenog dugotrajnog postupka hidrolize. Metoda je validirana prema ICH smjernicama i primijenjena u analizi pripravaka različitih dozirnih oblika Razvijena je i dvodimenzionalna metoda tekućinske kromatografije ultravisoke djelotvornosti spregnute s masenom spektrometrijom (2D-UHPLC-MS) za pouzdanu identifikaciju fitoestrogena iz skupine izoflavona u složenom matriksu ispitivanih uzoraka. Analiza pripravaka s fitoestrogenima namijenjenim ženama u postmenopauzi novom metodom pokazala je velika odstupanja između kromatografski određenog i deklariranog sadržaja ispitivanih izoflavona. The use of phytoestrogens has increased remarkably due to their biological activity and potential beneficial effects. An increasing number of dietary supplements containing different phytoestrogens are widely present in the market targeting postmenopausal women as a particular risk group. The content of isoflavones in dietary supplements not only differ quantitatively concerning their total isoflavone content, but also qualitatively concerning their isoflavone spectrum. The aim of the present study was to obtain data on pharmacokinetic properties for a set of 15 structurally related phytoestrogens (biochanin A, daidzein, formononetin, genistein, genistein-4,7-dimethylether, prunetin, 3,4,7-trihydroxyisoflavon, 4,6,7- trihydroxyisoflavon, 4,6,7-trimethoxyisoflavon, daidzin, genistin, ononin, sissotrin, coumestrol, coumestrol dimethylether) for the purpose of better understanding their biological activity and effects. Thereafter, based on the chromatographically determined lipophilicity, a novel, fast, selective and sensitive analytical method for simultaneous quantification of the 8 most important isoflavones in the aglycone and glycoside forms was developed. Lipophilicity of structurally related phytoestrogens was measured using reversed phase ultra-high performance liquid chromatography (RP-UHPLC). The phospholipid binding measurements were carried out using high performance liquid chromatography (HPLC) on a column with a stationary phase that contains phosphatidylcholine. The protein binding measurements were carried out using biomimetic chromatography. Caco-2 cells were used to estimate the intestinal permeability and the way of transport across the membrane of phytoestrogens test compounds. Obtained results differentiate phytoestrogens according to their structure where aglycones show significantly higher lipophilicity, IAM partitioning, AGP binding and Caco- 2 permeability comparing to glucosides. However, HSA binding was very high for all investigated compounds. Experimentally obtained pharmacokinetic parameters were compared with in silico predicted data by different computer platforms. A new RP-UHPLC method was developed for simultaneous determination of the most significant phytoestrogens from isoflavone family in aglycone (genistein, daidzein, biochanin A and formononetin) and glycoside (genistin, daidzin, sissotrin, ononin) forms without prior time-consuming process of hydrolysis. The method was validated as per ICH recommendations and was successfully applied to the analysis of various dietary supplements. Additionally to identification using external standard, the identity of selected compounds in samples was confirmed by using two-dimensional ultra-high performance liquid chromatography-tandem mass spectrometry (2D-UHPLC-MS). Analysis of dietary supplements containing phytoestrogens intended for use in postmenopausal women, using newly developed method, revealed significant deviations between measured and labelled content of investigated isoflavones. |
| Databáze: | OpenAIRE |
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