A Simple HPLC-UV Method For Simultaneous Determination of Levetiracetam and Carbamazepine

Abstract

ABSTRACT

A combination of levetiracetam (LEV) and carbamazepine (CBZ) may be used to

obtain synergistic effect in treatment of epilepsy. The aim of the study is to develop and

validate a simple reversed phase (RP) HPLC-UV method to determine simultaneously

LEV and CBZ in pure form and in different pharmaceutical formulations. Separation

was achieved on a C18 column (5 μm, 4.6 mm×250 mm) using a mobile phase

consisting of methanol and ultrapure water (45:55, v/v) at a flow rate of 1.0 mL/min

at 230 nm. Validation parameters (specificity, linearity, accuracy, precision, LOD and

LOQ) were studied according to the relevant ICH Guideline. The calibration curves

were linear over a concentration range of 0.5-32 μg/mL for both LEV and CBZ. The

intra- and inter-day accuracy values (as relative error; RE) were less than ±2% and also

the intra- and inter-day precision values (as the percent relative standard deviation;

RSD%) were less than 2%. According to relevant guidelines, the accuracy and precision

of the developed method is suitable. LOD and LOQ values were found to be 0.036 and

0.110 μg/mL for LEV and 0.026 and 0.078 μg/mL for CBZ, respectively. The simple,

specific, accurate, precise and sensitive analytical method developed could be useful for

simultaneous determination of LEV and CBZ.

Keywords: Carbamazepine, HPLC-UV method, Levetiracetam, Validation.

ÖZET

Epilepsi tedavisinde sinerjik etki elde etmek için levetirasetam (LEV) ve karbamazepin

(CBZ) birarada kullanılabilir. Bu çalışmanın amacı, saf halde ve farklı farmasötik

formülasyonlarda LEV ve CBZ’yi eş zamanlı olarak belirlemek üzere basit ters faz

(RP) HPLC-UV yönteminin geliştirilmesi ve geçerliliğinin gösterilmesidir. Ayırım, C18

kolonunda (5 μm, 4.6 mm x 250 mm), metanol ve ultra saf sudan (45:55, h/h) oluşan

hareketli faz kullanılarak 1.0 mL/dk akış hızında ve 230 nm’de gerçekleştirilmiştir.

Geçerlilik parametreleri (özgüllük, doğrusallık, doğruluk, kesinlik, LOD ve LOQ) ilgili

ICH Kılavuzuna göre çalışılmıştır Kalibrasyon eğrileri hem LEV hem de CBZ için

0.5-32 μg/mL derişim aralığında doğrusal olarak bulunmuştur. Gün içi ve günler arası

doğruluk değerlerinin [bağıl hata (BH) olarak] ±%2’den az ve gün içi ve günler arası

kesinlik değerlerinin de [% bağıl standart sapma; %BSS) %2’den az olduğu saptanmıştır.

İlgili kılavuzlara göre, geliştirilen yöntemin doğruluğu ve kesinliği uygundur. LOD ve

LOQ değerleri sırasıyla, LEV için 0.036 ve 0.110 μg/mL ve CBZ için sırasıyla 0.026

ve 0.078 μg/mL olarak bulunmuştur. Geliştirilen basit, özgül, doğru, kesin ve hassas

analitik yöntem LEV ve CBZ’nin eş zamanlı olarak saptanması için faydalı olabilir.

Anahtar Kelimeler: Geçerlilik, HPLC-UV yöntemi, Karbamazepin, Levetirasetam

Keywords

• Carbamazepine,HPLC-UV method,Levetiracetam,Validation

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