title: Development and Validation of a RP-HPLC Method for Determination of Related Substances and Degradants in Entacapone.
abstract: A new reverse phase-liquid chromatography (RP-HPLC) method has been developed for simultaneous determination of entacapone and its pharmacopoeia impurities, in-house impurities and degradation impurities (total 17 analytes). Chromatographic separation was achieved on a C18 column (size: 250 × 4.6 mm; 5 µm particle size) at a flow rate of 1.0 mL/min with 210 nm detection. The mobile phase (MP) consists of 1.361 g of potassium di-hydrogen phosphate and 1.742 g of di-potassium phosphate in 1.0 L water, pH adjusted to 2.5 with ortho phosphoric acid (MP-A) and acetonitrile (MP-B) through gradient elution. The product was subjected to stress conditions such as acid, base, peroxide, thermal and photolytic degradation. Two new impurities above 2% level were observed and isolated through preparative HPLC and well characterized. However, no interference observed due to degradation impurities and entacapone and its EP impurities, in-house impurities. As part of the method validation, specificity, limit of detection, limit of quantitation (LOQ), linearity, accuracy, precision, robustness and ruggedness were determined. LOQ values were achieved between 0.01 and 0.04%. Good linearity (r(2) > 0.99) was obtained ranging from LOQ to 150%. Recovery was verified for all impurities at concentrations ranging from LOQ to 150%. Hence, a newly developed RP-HPLC method was capable for well separation of all analytes with acceptable resolution and tailing factor.
authors.affiliation: Research and Development Centre, Suven Life Sciences Limited, Plot No.: 18, Phase-3; IDA, Jeedimetla, Hyderabad 500055, India purnachand_d@yahoo.com.
authors.forename: Dasari
authors.initials: D
authors.lastname: Purnachand
authors.affiliation: Research and Development Centre, Suven Life Sciences Limited, Plot No.: 18, Phase-3; IDA, Jeedimetla, Hyderabad 500055, India.
authors.forename: Arava
authors.initials: A
authors.lastname: Veerareddy
authors.affiliation: Department of Chemistry, JNT University, Hyderabad, India.
authors.forename: Bhoomireddy
authors.initials: B
authors.lastname: Ramadevi
authors.affiliation: Research and Development Centre, Suven Life Sciences Limited, Plot No.: 18, Phase-3; IDA, Jeedimetla, Hyderabad 500055, India.
authors.forename: Ch V S L
authors.initials: ChV
authors.lastname: Kameswarrao
authors.affiliation: Research and Development Centre, Suven Life Sciences Limited, Plot No.: 18, Phase-3; IDA, Jeedimetla, Hyderabad 500055, India.
authors.forename: Bethi
authors.initials: B
authors.lastname: Madhusudhanreddy
mesh_terms: D002396:Catechols / Q000737:chemistry* / Q000378:metabolism; D002623:Chemistry Techniques, Analytical / Q000379:methods* / Q000592:standards; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004340:Drug Contamination; D009570:Nitriles / Q000737:chemistry* / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D015203:Reproducibility of Results
chemical_list: D002396:Catechols; D009570:Nitriles; D004364:Pharmaceutical Preparations; C071192:entacapone