Method development and validation for the analysis of a new anti-cancer infusion solution via HPLC

• Published in: Journal of separation science Vol. 34; no. 2; pp. 135 - 141
• Main Authors: Donnarumma, Fabrizio, Schober, Margot, Greilberger, Joachim, Matzi, Veronika, Lindenmann, Jörg, Maier, Alfred, Herwig, Ralf, Wintersteiger, Reinhold
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley-VCH Verlag 01.01.2011; WILEY-VCH Verlag; WILEY‐VCH Verlag; Wiley; Wiley Subscription Services, Inc
• Subjects: Analysis; Antineoplastic Agents - analysis; Biological and medical sciences; Cancer; Chromatography; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; General pharmacology; High-performance liquid chromatography; HPLC; Hydroxymethylfurfural; Medical sciences; N-Acetyl-L-methionine; N-Acetyl-L-selenomethionine; pH effects; Pharmaceutical industry; Pharmacology. Drug treatments; Temperature effects; Wavelength; α-Ketoglutaric acid; Antineoplastic agent; Validation; Injectable form; Chemical analysis; N-Acetyl-L-methionine; HPLC chromatography; Furfural(hydroxymethyl); α-Ketoglutaric acid; Aldehyde; Glutaric acid(oxo); N-Acetyl-L-selenomethionine; Reversed phase chromatography; Ultraviolet detector; Hydroxymethylfurfural; Dosage form; Diode array; Ketoacid; HPLC; Quantitative analysis
• ISSN: 1615-9306; 1615-9314; 1615-9314
• DOI: 10.1002/jssc.201000574

A fast and simple HPLC method has been developed and validated for the quantification of a completely new anti-cancer drug during the manufacturing process. The combination of four compounds including α-ketoglutaric acid, hydroxymethylfurfural, N-acetyl-L-methionine and N-acetyl-L-selenomethionine, administered intravenously, is still in test phase but has already shown promising results in cancer therapy. HPLC separation was achieved on an RP-18 column with a gradient system. However, the highly different concentrations of the compounds required a variation in the detection wavelength within one run. In order to produce a chromatogram where peaks were comparable on a similar range scale, detection at absorption maxima for the two most concentrated components was avoided. After optimization of the gradient program it was possible to detect all four substances within 14 min in spite of their strongly different chemical structure. The method developed was validated for accuracy, repeatability, reproducibility and robustness in relation to temperature and pH of buffer. Linearity as well as the limit of detection and quantification were determined. This HPLC method was found to be precise, accurate and reproducible and can be easily used for in-line process control during the manufacture of the anti-tumour infusion solution.