Validation of an HPLC method for the simultaneous determination of eletriptan and UK 120.413

Abstract

Arapid and sensitive RPHPLCmethod was developed for the routine control analysis of eletriptan hydrobromide and its organic impurity UK 120.413 in Relpax® tablets. The chromatography was performed at 20°C using a C18 XTerra ™ (5 µm, 150 x 4,6 mm) column at a flow rate 1.0 ml/min. The drug and its impurity were detected at 225 nm. The mobile phase consisted of TEA (1 %) - methanol (67.2:32.8 v/v), the pH of which was adjusted to 6.8 with 85 % orthophosphoric acid. Quantification was accomplished by the internal standard method. The developed RP HPLC method was validated by testing: accuracy, precision, repeatability, specificity, detection limit, quantification limit, linearity, robustness and sensitivity. High linearity of the analytical procedure was confirmed over the concentration range of 0.05 - 1.00 mg/ml for eletriptan hydrobromide and from 0.10 - 1.50 µg/ml for UK 120.413, with correlation coefficients greater than r = 0.995. The low value of the RSD expressed the good repeatability and precision of the method. Experimental design and a response surface method were used to test robustness of the analytical procedure and to evaluate the effect of variation of the method parameters, namely the mobile phase composition, pH and temperature. They showed small deviations from the method setting. The good recovery and low RSD confirm the suitability of the proposed RP HPLC method for the routine determination of eletriptan hydrobromide and its impurity UK 120.413 in Relpax® tables.

U ovom radu je predstavljena brza i osetljiva RP HPLC metoda namenjena za rutinsko ispitivanje i kontrolu eletriptan hidrobromida i njegove organske nečistoće UK 120.413 u Relpax® tabletama. Hromatografski postupak je izveden uz korišćenje kolone C 18 XTerra ™ (5 µm, 150 x 4.6 mm) pri protoku mobilne faze od 1.0 ml/min i na 20 °C, a detekcija lekovite supstance i njene nečistoće je vršena na 225 nm. Mobilna faza se sastojala iz smeše TEA(1 %) - metanol (67,2:32,8 v/v), pH vodene faze je podešen na 6.8 sa 85 % ortofosfornom kiselinom. Kvantitativna analiza je vršena metodom internog standarda. Predložena RP HPLC metoda je validirana, a ispitivani su tačnost, preciznost, ponovljivost, specifičnost, limit detekcije, limit kvantifikacije, linearnost, robusnost i osetljivost metode. Visoka linearnost analitičkog postupka je potvrđena u opsegu koncentracija 0,05-1,00 mg/ml za eletriptan hidrobromid i 0,10-1,50 µg/ml za UK 120.413, sa koeficijentima korelacije koji su veći od r = 0,995. Niska vrednost relativne standardne devijacije potvrđuje dobru ponovljivost i preciznost metode. Eksperimentalni dizajn i metoda površine odgovora sistema su korišćeni u toku ispitivanja robusnosti da bi se procenio uticaj variranja vrednosti hromatografskih parametara metode. Test robusnosti je obuhvatao sastav mobilne faze, pH i temperaturu u malim variranjima vrednosti oko nominalne. Dobre "recovery" vrednosti i niska relativna standardna devijacija potvrđuju da je predložena RP HPLC pogodna za rutinsko određivanje eletriptan hidrobromida i njegove nečistoće UK 120.412 u Relpax® tabletama.