A novel method for the determination of chemical purity and assay of menaquinone-7. Comparison with the methods from the official USP monograph.

An HPLC method with UV detection and separation with the use of a C30 reversed phase analytical column for the determination of chemical purity and assay of menaquinone-7 (MK7) in one chromatographic run was developed. The method is superior to the methods published in the USP Monograph in terms of selectivity, sensitivity and accuracy, as well as time, solvent and sample consumption. The developed methodology was applied to MK7 samples of active pharmaceutical ingredient (API) purity, MK7 samples of lower quality and crude MK7 samples before purification. The comparison of the results revealed that the use of USP methodology could lead to serious overestimation (up to a few percent) of both purity and MK7 assay in menaquinone-7 samples.

Development and validation of high performance liquid chromatography with a spectrophotometric detection method for the chemical purity and assay of nepafenac.

The study is a proposition of the application of high performance liquid chromatography (HPLC) with a spectrophotometric UV range detector to analyze the chemical purity and assay of nepafenac, an active pharmaceutical ingredient (API). During literature search only a few publications were found about nepafenac. HPLC UV methods were mainly presented in patent documents about nepafenac synthesis and chemical purity. The presented method allows to separate all potential related compounds from nepafenac and to quantitate the nepafenac amount. As there is no official monograph in the pharmacopeias about nepafenac, the performed full validation procedure makes the method ready to use in routine analysis. The composition of the mobile phase (10mM ammonium formate, pH 4.1) and the HPLC column (Phenomenex Gemini-NX C18) were selected during the development step. Presented data confirm the benefits of the developed method. Four of the most potential impurities were validated as for the quantitative test and the rest of impurities were validated as for the limit test - according to ICH Q2(R1). The accuracy/recovery results for the chemical purity method are within 90-108%, in the case of assay studies from 99% to 101%; the limit of detection is as low as 15-30ng/mL. The linearity passes all statistical tests.

Identification and physicochemical characteristics of temozolomide process-related impurities.

In this article the crystal structures of the starting material TZ-5 and the key intermediate TZ-6 of temozolomide (TZ-7), an anticancer therapeutic agent, are presented, together with their spectroscopic and thermal characteristics. Both compounds crystallize in the triclinic P-1 space group. X-ray crystallography studies proved that the compound TZ-6 exists as a monohydrate. A complete structural assignment was obtained for the signals in the 1H-, 13C- and 15N-nuclear magnetic resonance spectra and the structures were confirmed by Fourier-Transform infrared and Raman spectroscopy. The article describes the importance of the high purity of TZ-6 during the small-scale plant production of TZ-7 in a desired polymorphic form III with the purity higher than 99.50%, according to an HPLC method.

Stability of sample solution as a crucial point during HPLC determination of chemical purity of temozolomide drug substance.

An HPLC method for determination of related substances in temozolomide drug substance was developed. Particular attention was paid to the stability studies due to the fact that temozolomide is unstable in a solution and quickly decomposes to its main degradation product 5-amino-4-imidazolecarboxamide (AIC). A mixture of diluted acetic acid and acetonitrile (4:1, v/v) as a diluent guaranteed lowering the decomposition of temozolomide in the solution. As it is not practically possible to fully eliminate the decomposition of temozolomide during an analysis, the mathematical correction of the results was proposed which allows to analyse almost five times more samples per week, comparing to the procedure without the application of the correction. The accuracy of the correction procedure was proved by investigating the recovery of AIC spiked to temozolomide solutions at different levels. Recoveries equalled 90-108% for AIC concentrations contained in the range of 0.30-1.80 µg ml(-1). The developed method was validated according to the current guidelines, proving the suitability of the method for its intended purpose.

Fractionation study in bioleached metallurgy wastes using six-step sequential extraction.

The stored metallurgy wastes contain residues from ore processing operations that are characterized by relatively high concentrations of heavy metals. The bioleaching process makes use of bacteria to recover elements from industrial wastes and to decrease potential risk of environmental contamination. Wastes were treated by solutions containing bacteria. In this work, the optimized six-stage sequential extraction procedure was applied for the fractionation of Ni, Cr, Fe, Mn, Cu and Zn in iron-nickel metallurgy wastes deposited in Southern Poland (Szklary). Fractionation and total concentrations of elements in wastes before and after various bioleaching treatments were studied. Analyses of the extracts were performed by ICP-MS and FAAS. To achieve the most effective bioleaching of Zn, Cr, Ni, Cu, Mn, Fe the usage of both autotrophic and heterotrophic bacteria in sequence, combined with flushing of the residue after bioleaching is required. 80-100% of total metal concentrations were mobilized after the proposed treatment. Wastes treated according to this procedure could be deposited without any risk of environmental contamination and additionally the metals could be recovered for industrial purposes.

Speciation analysis of arsenic in terrestrial plants from arsenic contaminated area.

Arsenic speciation analysis was carried out in plants collected from arsenic contaminated area. Two plant species were chosen for the investigation: Reed Grass (Calamagrostis arundinacea) and Lady Fern (Athyrium filix-femina). To characterize arsenic species several different extraction procedures were applied including enzymatic extraction and extraction using surfactant solution (SDS). Two-step sequential extraction (water+SDS) that assures the highest extraction efficiency was applied to extract arsenic species from plant material. HPLC with anion-exchange column was used to separate extracted arsenic compounds and ICP-MS was applied for quantitative arsenic determination after species separation.