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Title:Razvoj, validacija in primerjava bioanaliznih metod HPLC in LC-MS/MS za določanje kandesartana v humani plazmi
Authors:ID Forjan, Vanja (Author)
ID Brodnjak Vončina, Darinka (Mentor) More about this mentor... New window
ID Prosen, Helena (Comentor)
Files:.pdf MAG_Forjan_Vanja_2016.pdf (1,86 MB)
MD5: D8C4B5E1CAA1587D6DE2D1BE09065341
 
Language:Slovenian
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:Kandesartan je farmakološko aktivni metabolit kandesartan cileksetila iz skupine sartanov, ki se uporablja za zdravljenje hipertenzije. V okviru magistrske naloge smo najprej razvili metodo za določanje kandesartana v humani plazmi, ki obsega pripravo plazemskih vzorcev z ekstrakcijo na trdno fazo na ekstrakcijskih kolonah Strata X in analizo pripravljenih plazemskih vzorcev s HPLC instrumentalno tehniko. Razvito metodo smo validirali in dokazali, da je selektivna, točna, ponovljiva in linearna v območju od 4 ng/ml do 400 ng/ml. Da smo dosegli spodnjo mejo določljivosti 1 ng/ml, smo za detekcijo uporabili tandemsko masno spektrometrijo (LC-MS/MS) v SRM načinu z uporabo pozitivne ionizacije z elektro- razprševanjem. Po končanem razvoju smo metodo statistično ovrednotili v koncentracijskem območju od 1 do 400 ng/ml v humani plazmi in pri tem uporabili metodo internega standarda, ovrednotenje metode pa smo izvedli z določitvijo naslednjih validacijskih parametrov: selektivnost, linearnost, ponovljivost in pravilnost, spodnja meja določljivosti (LLOQ), izkoristek priprave vzorca, vpliv matrice, matrični faktor, vpliv redčenja, vpliv hemolitične plazme, testiranje čistosti sistema ter različne stabilnostne pogoje kandesartana v raztopinah in plazmi. Razviti in ovrednoteni metodi sta se v praksi izkazali kot učinkoviti, saj lahko z njima analiziramo veliko število vzorcev v kratkem času, hkrati pa sta natančni, točni in robustni v daljšem časovnem obdobju.
Keywords:kandesartan, SPE, HPLC, LC-MS/MS
Place of publishing:Maribor
Publisher:[V. Forjan]
Year of publishing:2016
PID:20.500.12556/DKUM-58437 New window
UDC:543.51:616.15(043.2)
COBISS.SI-ID:20380438 New window
NUK URN:URN:SI:UM:DK:UH8GDKQU
Publication date in DKUM:19.07.2016
Views:1908
Downloads:222
Metadata:XML RDF-CHPDL DC-XML DC-RDF
Categories:KTFMB - FKKT
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Secondary language

Language:English
Title:Development, validation and comparison of HPLC and LC-MS/MS bioanalytical methods for the determination of candesartan in human plasma
Abstract:Candesartan is a pharmacologically active metabolite of candesartan cilexetil from the sartan group, which is used to treat hypertension. A method for the determination of candesartan in human plasma, which includes the preparation of plasma samples by solid phase extraction using Strata-X extraction columns and analysis of prepared plasma samples by HPLC instrumental technique, was developed. The method was validated and was proved to be selective, accurate, repeatable, and linear in the range from 4 ng/mL to 400 ng/mL. In order to reach the lower limit of quantification of 1ng/ml, tandem mass spectrometry in the SRM mode with positive electrospray ionization was used for detection. When developed, the method was statistically validated in the concentration range of 1 – 400 ng/mL in human plasma, using the internal standard method. The validation of the method was performed by the evaluation of the following validation parameters: selectivity, linearity, repeatability and accuracy, lower limit of quantitative determination (LLOQ), recovery of sample preparation, matrix effects, matrix factor, effect of dilution and hemolysis, testing of system purity, and different stability conditions of candesartan in solutions and plasma. The developed and validated methods have proved to be effective in practice because they can be used for the analysis of a large number of samples in a short time. In addition, both methods are precise, accurate and robust over a longer period of time.
Keywords:candesartan, SPE, HPLC, LC-MS/MS


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