Evaluating phytochemical constituents and in-vitro antiplasmodial and antioxidant activities of Fadogiella stigmatoloba, Hygrophylla auriculata, Hylodesmum repandum, and Porphyrostemma chevalieri extracts.

Introduction: Fadogiella stigmatoloba, Hygrophylla auriculata, Hylodesmum repandum and Porphyrostemma chevalieri are used against malaria in traditional medicine in the Democratic Republic of the Congo (DRC). To evaluate their potential in the treatment of this disease, the in vitro antiplasmodial property of these four plants was evaluated. All experiments were conducted on methanolic extracts performed on selected organ parts of these plants. Methods: The methanolic extracts, obtained by maceration, were firstly screened in vitro against the chloroquine sensitive (3D7) and resistant (W2) Plasmodium falciparum strains by the measurement of lactate dehydrogenase activity, and on human keratinocytes (HaCat) cells by the MTT assay to determine their selectivity indices (SI). Secondly, the antioxidant activity of the same extracts was evaluated using DPPH and FRAP assays. Finally, the presence of specific phytochemical constituents was evaluated using standard methods and tentatively identified by GC-MS. Results: An optimum antiplasmodial activity (IC50 = 3.4 ± 0.7 µg/mL, for 3D7, SI = 58.2; IC50 = 7 ± 1.0 µg/mL, for W2, SI = 28.3) was obtained with the leave extract of P. chevalieri. The leaves (for F. stigmatoloba and H. repandum), and the aerial part (for H. repandum) extracts showed promising and moderate antiplasmodial activities against respectively the 3D7 strain (IC50: <15 µg/mL), and W2 strain (IC50:15-50 µg/mL). All extracts presented a weak cytotoxic effect (IC50: >100 µg/mL) on HaCat cells. For the antioxidant test, the most interesting activity was obtained with the leaf extract of P. chevalieri. The GC-MS analysis of these four plants species extracts revealed the presence of various compounds, such as Ethyl 2-nonenoate, 2-(2-Hydroxy-2-phenylethyl)-3,5,6-trimethyl pyrazine, Palmitic Acid, Ethyl palmitate, Ethyl linolenate, and N-Acetyltyramine. Conclusion: Based on the obtained results, P. chevalieri could be selected for further investigations or /and for the management of malaria after standardization.

Proposals for Antimicrobial Testing Guidelines Applied on Ajowan and Spanish Lavender Essential Oils.

To fight the rising resistance of microorganisms to antibiotics, a strategy followed by several researchers is to focus on natural compounds, such as essential oils, as a source of potent antibacterial compounds. These last decades, hundreds of original papers have been written about microbiological assays that prove the antibacterial activity of essential oils and their use in the medical field. But can we really compare all the data available in the literature when the raw material, the microbiological assays, and/or the strains are different from one article to another? This review will point out the differences and the inadequate practices found in published articles that tested 2 lesser-studied essential oils-Spanish lavender and the ajowan-by the broth dilution method against Staphylococcus aureus, a human pathogenic bacterium. Many pitfalls were found in the literature, for example, a variable chemical composition rarely underlined by the authors, unidentified strains or clinical strains used without a related antibiogram, a lack of quality controls, and the assertion of questionable positive results. At last, some general guidelines that should be followed by every scientific researcher will be discussed.

Rapid method for the confirmatory analysis of chrysoidine in aquaculture products by ultra-performance liquid chromatography-tandem mass spectrometry.

A sensitive and fast method for the quantification of the illegal dye chrysoidine in aquaculture products with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) is presented. Muscle tissues were made alkaline with sodium hydroxide and extracted with ethyl acetate. After evaporation and subsequent defatting with n-hexane, extracts were directly injected onto the UPLC-column. Chromatography was performed on a C(18) column using 0.1% formic acid in water and an acetonitrile gradient within 6 min. Mass spectrometric analysis was performed in the positive electrospray MS/MS mode. The limit of quantification was 0.25 ng/g, which was 30 times lower than the only previously published method with gas chromatographic detection. A complete validation according to the scientific literature and as defined by the European Union was performed. The applicability of the method was shown in the analysis of more than 50 unknown samples in the framework of a monitoring program.

Validation of a method for the detection and confirmation of nitroimidazoles and the corresponding hydroxy metabolites in pig plasma by high performance liquid chromatography-tandem mass spectrometry.

Nitroimidazoles (Ronidazole, Dimetridazole, Metronidazole, Ipronidazole) and their hydroxy metabolites are banned substances with antibiotic and anticoccidial activity. They are suspected to be carcinogenic and mutagenic. Since nitroimidazoles showed an inhomogeneous distribution and a rapid degradation in incurred muscle samples, plasma is the preferred target matrix for residue analysis. The analytical method of Polzer et al. [J. Polzer, C. Stachel, P. Gowik, Anal. Chim. Acta 521 (2004) 189] was adapted for liquid chromatography-tandem mass spectrometry detection and was validated in house according to the Commission Decision 2002/657/EC. The method is specific for all nitroimidazole except for Ipronidazole and its metabolite, due to interferences at their retention times in chromatograms of blank plasma and reagents samples. The absence of a matrix effect enables the use of a (linear) calibration curve in solution for quantitation. The apparent recovery (obtained after correction with a deuterated internal standard) is between 93% and 123%, except for the metabolite of Metronidazole (58-63%). The repeatability (CVr=2.49-13.39%) and intralaboratory reproducibility (CVRW=2.49-16.38%) satisfy the Horwitz equation. The obtained values for the detection capacity (CCbeta) range from 0.25 to 1 microg L(-1), while values obtained for the decision limit (CCalpha) are below CCbeta.

Determination of acrylamide levels in selected foods in Brazil.

Selected carbohydrate-rich foods available on the Brazilian market (111 samples representing 19 product categories) were analysed for acrylamide content using a liquid chromatography-tandem mass spectrometry method. A limit of detection of 10 microg kg(-1), a limit of quantitation of 20 microg kg(-1) and mean recoveries ranging 100 to 115% were obtained during a laboratory validation procedure. The concentration of acrylamide in the samples ranged from <20 to 2528 microg kg(-1), with a considerable variation between individual foodstuffs within the same food product class. The highest levels were found in potato products processed at high temperature and in instant coffee, while the lowest concentrations were detected in cassava- and maize-based products, bread and beer. These results are comparable with those reported in other countries.

Selection criteria for potato tubers to minimize acrylamide formation during frying.

A number of parameters linked to the selection of potato tubers were evaluated with regard to their potential to influence acrylamide formation in French fries. The formation of acrylamide, which is a potential human carcinogen, can be minimized for a big extent by the selection of an appropriate tuber. This study focused on the following selection criteria: variety as influenced by storage time and soil type, underwater weight, and tuber size. A total of 16 varieties were compared, concerning their potential for acrylamide formation. From that survey, certain varieties, such as Tebina and Quincy, could be appointed as unsuitable for frying. The differences in the potential of acrylamide formation between the varieties could mainly be explained by the reducing sugar content of the potato (R2 = 0.82, n = 96). The investigated type of soil and storage time at 8 degrees C appeared to have a minor influence on the acrylamide formation during frying. On the other hand, the tuber size of the potato did contribute in a significant manner to the acrylamide formation. Smaller tubers were more susceptible to acrylamide formation and should be avoided in the frying process. The last selection parameter, the underwater weight, appeared to be of minor importance in the acrylamide formation. On the basis of these simple selection criteria, it is possible to make a first screening of potatoes to reduce the acrylamide formation during frying.

Influence of fertilization on acrylamide formation during frying of potatoes harvested in 2003.

The quality of the potato has been found to vary, when grown under different agricultural and environmental conditions, such as the level of fertilization. Consequently these factors may influence the acrylamide formation during the preparation of French fries. These assumptions were studied on three varieties: Bintje, Ramos, and Saturna from the harvest of 2003. Decreasing N fertilization caused increases in the reducing sugar concentration from 60% up to 100% on DM for all varieties studied. Due to a high correlation between the reducing sugar content and the generation of acrylamide during frying, this resulted in a parallel increase in the acrylamide concentration of the French fries. Thus by lowering the amount of N fertilizer, an increase of 30-65% of the acrylamide generation during frying could be observed. It seems of extreme importance to find an appropriate balance between the level of N fertilizer in order to diminish acrylamide formation but on the other hand to obtain an acceptable tuber and to consider the environmental impact. All results reported should be seen in the perspective of the warm growing season of 2003.

Influence of storage practices on acrylamide formation during potato frying.

A number of parameters linked to storage of potatoes were evaluated with regard to their potential to influence the acrylamide formation in French fries. Acrylamide, which is a potential human carcinogen, is reported to be formed during the frying of potatoes as a result of the reactions between asparagine and reducing sugars. This study was conducted using three potato varieties (Bintje, Ramos, and Saturna) typically used in Belgium, The Netherlands, and the northern part of France for French fry and crisp production. Saturna, mainly used in crisp production, appeared to be the least susceptible for acrylamide formation during frying. Especially storage at low temperatures (4 degrees C) compared to storage at 8 degrees C seemed to enhance acrylamide formation due to a strong increase in reducing sugars caused by low-temperature storage. Because of the reversible nature of this physiological reaction, it was possible to achieve a significant reduction of the reducing sugars after a reconditioning of the cold-stored potatoes for 3 weeks at 15 degrees C. All changes in acrylamide concentrations could mainly be explained by the reducing sugar content of the potato (R2 = 0.84, n = 160). This means that, by ensuring a low reducing sugar content of the potato tuber, the risk for acrylamide formation will largely be reduced. Finally the use of a sprout inhibitor did not influence the composition of the potato, and thus acrylamide formation was not susceptible to this treatment.