DNA extracted from plants rich in polyphenols and/or polysaccharides is often problematic when subjected to polymerase chain reaction, especially when mature tissues are used for DNA extraction. In order to overcome the problems associated with poor-quality DNA extracted from such plant samples, a protocol has been developed, availing on a high salt concentration and on the combination of polyvinylpyrrolidone and activated charcoal in the extraction buffer, in order to prevent the solubilization of polysaccharides and polyphenols in the DNA extract. Mild temperature conditions during extraction and precipitation were also recognized as important parameters. Besides DNA purity, mild precipitation conditions were found to be beneficial in obtaining less low-molecular mass nucleic acids in the final DNA extract. The homogenization step and the amount of sample extracted were also found to be crucial in keeping the extraction procedure robust.
We developed a simple and robust liquid chromatographic/mass spectrometric method (LC-MS) for the quantitative analysis of 10 sterols from the late part of cholesterol synthesis (zymosterol, dehydrolathosterol, 7-dehydrodesmosterol, desmosterol, zymostenol, lathosterol, FFMAS, TMAS, lanosterol, and dihydrolanosterol) from cultured human hepatocytes in a single chromatographic run using a pentafluorophenyl (PFP) stationary phase. The method also avails on a minimized sample preparation procedure in order to obtain a relatively high sample throughput. The method was validated on 10 sterol standards that were detected in a single chromatographic LC-MS run without derivatization. Our developed method can be used in research or clinical applications for disease-related detection of accumulated cholesterol intermediates. Disorders in the late part of cholesterol synthesis lead to severe malformation in human patients. The developed method enables a simple, sensitive, and fast quantification of sterols, without the need of extended knowledge of the LC-MS technique, and represents a new analytical tool in the rising field of cholesterolomics.
An original gas chromatographic method has been developed for simultaneous determination of major terpenes and cannabinoids in plant samples and their extracts. The main issues to be addressed were the large differences in polarity and volatility between both groups of analytes, but also the need for an exhaustive decarboxylation of cannabinoid acidic forms. Sample preparation was minimised, also by avoiding any analyte derivatisation. Acetone was found to be the most appropriate extraction solvent. Successful chromatographic separation was achieved by using a medium polarity column. Limits of detection ranged from 120 to 260 ng/mL for terpenes and from 660 to 860 ng/mL for cannabinoids. Parallel testing proved the results for cannabinoids are comparable to those obtained from established HPLC methods. Despite very large differences in concentrations between both analyte groups, a linear range between 1 and 100 µg/mL for terpenes and between 10 and 1500 µg/mL for cannabinoids was determined.
5 August 2019 in European Food Research and Technology
Floral nectar is mainly a reward in the form of food for pollinators. Its composition plays an important role when pollinators choose their food. Several studies have shown that the popularity of flowers with nectar is influenced by the concentration and ratio of sugars. Here, we present the nectar chemical composition with regard to three main sugars and their concentrations in correlation with abiotic factors for the plant species Salvia pratensis L. and Salvia glutinosa L. through their 2023 flowering season. We sampled nectar using microcapillaries at three different times during the day on sites in nature. Our results show that nectar production in both species is the highest at around 12 a.m. The abiotic factor that affects nectar production in both species is the soil temperature, while UVB radiation does not influence nectar production. Air temperature and air humidity affect the nectar production of S. glutinosa, while soil humidity affects nectar production in S. pratensis. The most represented sugar in S. glutinosa nectar is sucrose, while S. pratensis nectar has more glucose and fructose. Our results show that UVB radiation has an effect on the sucrose level, although it does not have any direct effect on nectar productivity.