HPLC-UV Method Development and Validation for the Determination of Low Level Formaldehyde in a Drug Substance

• Published in: Journal of chromatographic science 2008-07, Vol.46 (6), p.461-465
• Main Authors: Soman, A., Qiu, Y., Chan Li, Q.
• Format: Article
• Language: English
• Subjects: Analysis; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Formaldehyde - analysis; General pharmacology; Kinetics; Medical sciences; Pharmaceutical Preparations - chemistry; Pharmacology. Drug treatments; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet - methods
• ISSN: 0021-9665; 1945-239X
• DOI: 10.1093/chromsci/46.6.461

A reversed-phase high-performance liquid chromatographic method (HPLC) with diode-array detection is developed and validated for the quantitative determination of formaldehyde in a drug substance. Formaldehyde (HCHO) is reacted with 2,4- dinitrophenylhydrazine (DNPH) to form a Schiff base (HCHODNPH derivatization product), which has an absorbing maximum (λmax) at 360 nm. The HPLC method employs a C8, 3-µm particle size analytical column (150 mm × 4.6 mm), 15-µL injection volume, column temperature controlled at 30°C, detection at 360 nm, and a water-acetonitrile (55:45, v/v) mobile phase at a flow rate of 1 mL/min. These conditions resolve the HCHO-DNPH product from unreacted DNPH, the drug substance and related impurities, as well as diluent peaks within 20 min. The retention time of the HCHO-DNPH product is approximately 6.4 min. The method is linear, accurate in the specified range (0.33–333 ppm), and robust based on analyte (HCHO-DNPH derivatization product) stability in standard and sample. Detection limit is 0.03 ng (0.1 ppm).