Razvoj analizne metode za določevanje alkil halidov s plinsko kromatografijo v aktivni farmacevtski učinkovini

diplomsko delo visokošolskega strokovnega študijskega programa I. stopnje

Damir Orkić (Avtor), Darinka Brodnjak-Vončina (Mentor), Mitja Kolar (Komentor)

Povzetek

Glavna naloga farmacevtske industrije je pacientu zagotoviti varno in kvalitetno izdelano zdravilo. V zadnjem času se veliko pozornosti posveča kontroli potencialnih genotoksičnih nečistoč (PGI), saj le-te povzročajo genske mutacije, kromosomske prelome ter preureditve, ki lahko vodijo v rakava obolenja. Dovoljena meja za vsebnost PGI-jev v zdravilih je običajno zelo nizka (koncentracijski nivo µgg-1) in je odvisna od dnevnega odmerka. V skupino teh nečistoč uvrščamo tudi alkil halide, ki lahko nastajajo kot stranski produkti različnih sinteznih stopenj, postopkov čiščenja (tvorba API soli) ali pa so kot reagenti prisotni v sintezi. Cilj dela je bil razvoj občutljive in selektivne analizne metode, ki bo direktno omogočala določevanje alkil halidov (klorometana, kloroetana bromometana in bromoetana) v aktivni farmacevtski učinkovini (API). Zaradi izredne hlapnosti analitov smo se razvoja analizne metode lotili s tehniko plinske kromatografije. Za uspešno ločbo analitov smo z neposrednim injiciranjem standardnih raztopin v metanolu preizkusili polarne, srednje-polarne in tudi nepolarne »WCOT« kapilarne kolone v kombinaciji z različnimi detektorji. Največji izziv je bilo izbrati kolono, ki bo zagotovila utrezno ločitev med analiti in topilom (metanol). Ločitev smo dosegli na koloni DB-624 v kombinaciji z masno-selektivnim detektorjem (MSD). Po večkratnem neposrednem injiciranju raztopine vzorca smo ugotovilili slabo ponovljivost, nestabilnost bazne linije in slabo obliko kromatografskih vrhov. Neposredno injiciranje smo nato uspešno nadomestili z injiciranjem plinaste faze s tehniko nadprostora (HS - headspace). Analizno metodo smo optimirali in jo tudi validirali kot limitni test na nivoju 2 µgg-1 (µgml-1). Pri validaciji smo potrdili selektivnost analizne metode, ponovljivost injiciranja, ponovljivost metode (za vzorec in vzorec s standardnim dodatkom), točnost ter robustnost. Z metodo smo nato analizirali realne vzorce izdelanega API-ja in potrdili ustreznost le-tega (vsebnost vseh analiziranih alkil halidov je bila pod mejo poročanja). Dobljeni rezultati so uradni del kontrolne strategije in so vključeni v registracijsko dokumentacijo.

Ključne besede

genotoksične nečistoče;alkil halidi;aktivna farmacevtska učinkovina;plinska kromatografija;masno delektivni detektor;validacija;

Podatki

Jezik:	Slovenski jezik
Leto izida:	2015
Tipologija:	2.11 - Diplomsko delo
Organizacija:	UM FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik:	[D. Orkič]
UDK:	543.544.3:661.12(043.2)
COBISS:	18667798
Št. ogledov:	1690
Št. prenosov:	244
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	DEVELOPMENT OF ANALYTICAL METHOD FOR THE DETERMINATION OF ALKYL HALIDES BY GAS CHROMATOGRAPHY IN ACTIVE PHARMACEUTICAL INGREDIENT
Sekundarni povzetek:	The main focus of Pharmaceutical industry is to produce safe and quality manufactured drug substance, which will ensure patient's safety. Recently, much attention is paid to the control of potential genotoxic impurities (PGI), since these cause gene mutations, chromosomal breaks and rearrangements, which may lead to cancer. Permissible limit for the content of PGI's in medicines is usually very low (concentration level mgkg-1) and it depends on the daily dose. One group of these impurities are also alkyl halides, which may be formed as by-products of the various synthetic steps, cleaning processes (the formation of API salts) or they can be present as reagents in the synthesis. The aim of my work was to develop a sensitive and selective analytical method that will allow direct determination of alkyl halides (methyl chloride, ethyl chloride, methyl bromide and ethyl bromide) in active pharmaceutical ingredient (API). Because of the extremely volatile nature of analytes method was developed by using gas chromatography (GC) as an analytical technique. To confirm the appropriate separation of the analytes and solvent (methanol), a direct injection of standard solutions in methanol was performed. During this process we tested polar, medium polar and non-polar "WCOT" capillary columns in combination with a variety of GC detectors. The biggest challenge was to identify the column, which would provide suitable separation between the analytes and the solvent (methanol). Appropriate separation was obtained on GC column DB-624 in combination with mass-selective detector (MSD). After repeated direct injections of the sample solution poor repeatability, baseline instability and poor shape of peaks were identified. Direct injection was then successfully replaced by injecting a gas phase with headspace injection (HS). Analytical method was optimized and validated as a limit test at level 2 µgg-1 (µgml-1). During validation of the analytical method selectivity, reproducibility of injections, reproducibility (for pure and spiked sample), accuracy and robustness were confirmed. The method was then used for testing of real samples of produced API. It was shown that the assay of all analyzed alkyl halides was below reporting limit. The results are an official part of the control strategy and are included in the registration dossier (DMF).
Sekundarne ključne besede:	genotoxic impurities;alkyl halides;active pharmaceutical ingredients;gas chromatography;mass selective detector;validation;
URN:	URN:SI:UM:
Vrsta dela (COBISS):	Diplomsko delo/naloga
Komentar na gradivo:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Strani:	XI, 77 f.
ID:	8750988