Quality of Non-Sterile Herbal Pharmaceutical Products – Detection of Fungi

The aim of this paper was to use real-time quantitative PCR (qPCR) to perform microbiological purity tests, aiming to identify the presence/absence of fungal DNA and its quantification from non-sterile pharmaceuticals, with different manufacturers and various compositions. For identification a potential presence of fungi in non-sterile drug samples, fungal DNA has been isolated, amplified and quantified under real-time qPCR conditions. qPCR analysis of non-sterile pharmaceutical samples showed the presence of fungi in all investigated non-sterile herbal drugs. The highest amount of fungal DNA was recorded for S2 sample (0.0767 ng) and the lowest fungal DNA quantities have been registered for S1 sample (0.000032 ng). qPCR-based methods provided an earlier and more sensitive detection, identification, and quantification of fungal contamination compared to standard methods

HPLC Method Validation for Simultaneous Determination of Three Mycotoxins from Corn Seeds

A new HPLC optimized method for simultaneous determination of some mycotoxins by separation and using a Dionex UltiMate3000 modular system, with multichannel UV detector was developed and validated. The method optimization was performed to determine simultaneously, several relevant mycotoxins from corn seeds that were stored over 8 months. The mycotoxins selected for quantification were: aflatoxin B1, ochratoxin A and zearalenone. They were selected due to their high concentration in the fodder. For the development stages of the HPLC method, it was introduced an internal standard to have accurate results. Taking into account that the analytes should be extracted from the complex matrix they reside, an extraction procedure was performed, using organic solvents, and the selection after repeated tests demonstrated the best capacity for ethyl acetate. The recovery is about 60–70 % after the extraction process also there is a good preconcentration (×2.5) of the analytes and the internal standard for their quantification. The limit of quantification (LOQ) obtained by chromatographic parameters optimization, for aflatoxin B1 and ochratoxin A were about 3–5 μg kg–1 and 14.4 μg kg–1 for zearalenone of raw biological material, making these values lower than those accepted by the actual normatives and regulations. (doi: 10.5562/cca1788