Determination of Potential Genotoxic Impurity, 5-Amino-2-Chloropyridine, in Active Pharmaceutical Ingredient Using the HPLC-UV System

Abstract

A novel analytical method, based on high-performance liquid chromatography with a UV (HPLC-UV) detection system for the sensitive detection of a genotoxic impurity (GTI) 5-amino-2-chloropyridine (5A2Cl) in a model active pharmaceutical ingredient (API) tenoxicam (TNX), has been developed and validated. The HPLC-UV method was used for the determination of GTI 5A2Cl in API TNX. The compounds were separated using a mobile phase composed of water (pH 3 adjusted with orthophosphoric acid): MeOH, (50:50: v/v) on a C18 column (150 × 4.6 mm i.d., 2.7 μm) at a flow rate of 0.7 mL min−1. Detection was carried out in the 254 nm wavelength. Column temperature was maintained at 40°C during the analyses and 10 μL volume was injected into the HPLC-UV system. The method was validated in the range of 1–40 μg mL−1. The obtained calibration curves for the GTI compound was found linear with equation, y = 40766x − 1125,6 (R2 = 0.999). The developed analytical method toward the target compounds was accurate, and the achieved limit of detection and limit of quantification values for the target compound 5A2Cl were 0.015 and 0.048 μg mL−1, respectively. The recovery values were calculated and found to be between 98.80 and 100.03%. The developed RP-HPLC-UV analytical method in this research is accurate, precise, rapid, simple and appropriate for the sensitive analysis of target GTI 5A2Cl in model API TNX.

Eco-Friendly High-Performance Thin-Layer Chromatography Method for the Determination of Tenoxicam in Commercial Formulations

“…Some high-performance liquid chromatography (HPLC) approaches were utilized to determine TNX in pure and dosage forms. − In the transdermal therapeutic system, TNX and meloxicam were also determined using an HPLC approach . A HPLC approach has also been used to measure potential genotoxic impurity, 5-amino-2-chloropyrine in TNX . Numerous HPLC assays were also used for the determination of TNX in the plasma samples of human subjects. − The liquid-chromatography mass-spectrometry (LC-MS) and micro-HPLC methodologies were utilized to identify TNX in conjunction with other NSAIDs in pharmaceutical preparations and biological materials, including blood, plasma, and erythrocytes .…”

“… 17 A HPLC approach has also been used to measure potential genotoxic impurity, 5-amino-2-chloropyrine in TNX. 18 Numerous HPLC assays were also used for the determination of TNX in the plasma samples of human subjects. 19 − 23 The liquid-chromatography mass-spectrometry (LC-MS) and micro-HPLC methodologies were utilized to identify TNX in conjunction with other NSAIDs in pharmaceutical preparations and biological materials, including blood, plasma, and erythrocytes.…”