Malaysian Journal of Analytical Sciences Vol 19 No 3 (2015): 595 – 602

 

 

 

DEVELOPMENT AND VALIDATION OF A STABILITY-INDICATING RP-HPLC METHOD FOR DETERMINATION OF AMBRISENTAN IN BULK DRUGS

 

(Pembangunan dan Validasi Kaedah KCPT-Fasa Terbalik yang Stabil bagi Penentuan Ambrisentan  di dalam Dadah Pukal)

 

Mohammed Nazeerunnisa¹, Lakshmi Garikapati², Syama Sundar Bethanabhatla^1,3*

 

¹Department of Chemistry,

Acharya Nagarjuna University, Nagarjuna Nagar-522510, India

² Centre for Pharmaceutical Sciences,

Jawaharlal Nehru Technological University, Hyderabad – 500085, India.

³Vice Chancellor,

Yogi Vemana University, Kadapa-516003, India.

 

*Corresponding author: profbsyamsundar@yahoo.co.in

 

 

Received: 18 February 2015; Accepted: 17 April 2015

 

 

Abstract

A stability-indicating reversed phase high-performance liquid chromatographic (RP-HPLC) method was developed for quantitative determination of ambrisentan and its potential related substances in bulk drugs. Drug substance was subjected to various stress conditions such as hydrolysis, oxidation, photolysis and thermal degradation as per International Conference on Harmonization (ICH) guidelines to investigate the stability indicating nature of the method. Significant degradation was found in acidic stress conditions. Efficient chromatographic separation was accomplished on a Phenomenex Luna C₁₈ (250 × 4.6 mm, 5 µm) column with a mobile phase composed of 0.02 M ammonium acetate buffer (pH = 4.2), and acetonitrile in 52:48 (v/v) ratio at a flow rate of 1.0 mL/min and column temperature of 25 ˚C. The eluents were monitored with a photo diode array (PDA) detector at a wavelength of 215 nm. The developed liquid chromatographic method was validated with respect to linearity, accuracy, precision, limits of detection and quantitation, and robustness. Regression analysis showed correlation co-efficient values greater than 0.997 for ambrisentan and its five impurities. Accuracy of the method was established based on the recovery obtained between 92.8 – 103.5% and 99.2 – 100.7% for impurities and ambrisentan, respectively.

 

Keywords: ambrisentan, RP-HPLC, forced degradation, stability indicating, validation

 

Abstrak

Kestabilan kaedah kromatografi cecair berprestasi tinggi - fasa terbalik (KCPT-Fasa Terbalik) telah dibangunkan bagi penentuan kuantitatif terhadap ambrisentan dan potensi bahan berkaitannya di dalam dadah pukal. Peubahan dadah adalah tertakluk kepada pelbagai keadaan tekanan seperti hidrolisis, pengoksidaan, fotolisis dan degradasi terma seperti dinyatakan di dalam garis panduan International Conference on Harmonization (ICH) bagi menkaji kaedah kestabilan. Kemerosotan yang ketara telah ditemui dalam keadaan tekanan berasid. Pemisahan kromatografi yang berkesan telah dicapai oleh turus Phenomenex Luna C18 (250 × 4.6 mm, 5 μm) dengan fasa bergerak terdiri daripada 0.02 M larutan penampan ammonium asetat (pH = 4.2), dan asetonitril dalam nisbah 52:48 (v/v) pada kadar aliran 1.0 mL/min dan suhu turus adalah 25 ˚C. Eluen dipantau oleh pengesan photo diode array (PDA) pada panjang gelombang adalah 215 nm. Kaedah kromatografi cecair yang dibangunkan kemudian telah ditentusahkan terhadap kelinearan, ketepatan, ketepatan, had pengesanan dan kauntifikasi, dan keteguhan. Analisis regresi menunjukkan nilai korelasi bersama mencapai 0.997 untuk ambrisentan dan lima bendasing. Ketepatan kaedah yang dibangunkan berdasarkan perolehan semula masing – masing di antara 92.8 – 103.5% dan 99.2 – 100.7% untuk bendasing dan ambrisentan.

 

Kata Kunci: ambrisentan, RP-HPLC, degradasi paksa, petunjuk kestabilan, validasi

 

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