Sequential extraction of carnosic acid, rosmarinic acid and pigments (carotenoids and chlorophylls) from Rosemary by online supercritical fluid extraction-supercritical fluid chromatography.

A high degree of selectivity is required during the plant extraction process in order to obtain extracts enriched in specific compounds or to avoid the extraction of unwanted ones. Rosemary is well known for its antioxidant compounds (carnosic acid, carnosol and rosmarinic acid). The plant also contains pigments (i.e. carotenoids, chlorophylls) which may cause a colour problem during the use of the extract in cosmetic formulations, for example. Supercritical fluid extraction is considered as a selective technique for plant extraction. Due to the physico-chemical properties of supercritical fluids, related to pressure, temperature and modifier addition, it is possible to carry out sequential extraction with successive conditions to collect different fractions that are rich either in pigments or in bioactive compounds. The aim of this study was to selectively extract bioactive compounds (i.e. carnosic acid and rosmarinic acid) and pigments (carotenoids and chlorophylls) from rosemary using supercritical fluid extraction. The optimisation of the extraction method was carried out using supercritical fluid extraction online coupled with a supercritical fluid chromatography (SFE-SFC) system. Two columns of different polarities were coupled to achieve the separation of the targeted compounds every five minutes during the extraction. Four fractions were obtained: a first one rich in carotenoids obtained with pure CO2 (25°C and 20 MPa), a second rich in carnosic acid obtained with 3% polar modifier (EtOH:water 50/50 v/v), a third fraction rich in rosmarinic acid using 10% of the same modifier and a fourth fraction rich in chlorophylls with 30% of ethanol as modifier. These four samples were then analysed by UHPLC-DAD-ESI-QTOF-HRMS in order to identify other extracted compounds and to study how the selected conditions impacted their extraction.

Selective extraction of bioactive compounds from plants using recent extraction techniques: A review.

Plant extraction has existed for a long time and is still of interest. Due to technological improvements, it is now possible to obtain extracts with higher yields. While global yield is a major parameter because it assesses the extraction performance, it can be of interest to focus on the extraction of particular compounds (specific metabolites) to enrich the sample and to avoid the extraction of unwanted ones, for instance the primary metabolites (carbohydrates, triacylglycerols). The objective then is to improve extraction selectivity is then considered. In solid-liquid extraction, which is often called maceration, the solvent has a major impact on selectivity. Its polarity has a direct influence on the solutes extracted, related to the chemical structure of the compounds, and modelling compound/solvent interactions by using various polarity or interaction scales is a great challenge to favor the choice of the appropriate extracting liquid. Technical advances have allowed the development of recent, and sometimes green, extraction techniques, such as Microwave-Assisted Extraction (MAE), Ultrasound-Assisted Extraction (UAE), Pressurized Liquid Extraction (PLE) and Supercritical Fluid Extraction (SFE). This review focuses on the specificity of these recent techniques and the influence of their physical parameters (i.e. pressure, intensity, etc.). In addition to the solvent selection, which is of prime interest, the physical parameters applied by the different techniques influence the extraction results in different ways. Besides, SFE is a versatile and green technique suitable to achieve selectivity for some compounds. Due to its properties, SC-CO2 allows tailoring conditions to improve the selectivity.

Supercritical Fluid Chromatography development of a predictive analytical tool to selectively extract bioactive compounds by supercritical fluid extraction and pressurised liquid extraction.

Selective extraction is a great concern in the field of natural products. The interest is to apply specific conditions favouring the solubility of targeted secondary metabolites and avoiding the simultaneous extraction of unwanted ones. Different ways exist to reach selective extractions with suited conditions. These conditions can be determined from experimental studies through experimental design, but a full experimental design takes time, energy, and uses plant samples. Prediction from varied solubility models can also be applied allowing a better understanding of the final selected conditions and eventually less experiments. The aim of this work was to develop and use a chromatographic model to determine optimal extraction conditions without the need for numerous extraction experiments. This model would be applied on the selective extraction of the desired antioxidant compounds in rosemary leaves (rosmarinic and carnosic acids) vs chlorophyll pigments to limit the green colour in extracts. This model was achieved with Supercritical Fluid Chromatography (SFC) and then applied to Supercritical Fluid Extraction (SFE) and Pressurised Liquid Extraction (PLE) assays. SFC models predicted low solubility of chlorophylls for low (5%) and high (100%) percentage of solvent in carbon dioxide. Also, low solubility was predicted with acetonitrile solvent compared to methanol or ethanol. This was confirmed with different extractions performed using SFE with different percentages of solvent (5, 30, and 70%) and with the three solvents used in the SFC models (acetonitrile, methanol and ethanol). Also extractions using PLE were carried out using the same neat solvents in order to confirm the SFC models obtained for 100% of solvent. Globally, extractions validated the SFC models. Only some differences were observed between ethanol and methanol showing the complexity of plant extraction due to matrix effect. For all these extracts, the content of carnosic acid and rosmarinic acid was also monitored and selective extraction conditions of bioactive compounds could be determined.

Evaluation of the extraction and stability of chlorophyll-rich extracts by supercritical fluid chromatography.

In this study, a rapid (less than 10 min) analytical method by reverse-phase supercritical fluid chromatography was developed with an isocratic mobile phase, enabling the separation of 11 compounds, chlorophyll a and b, pheophytin a and numerous allomers or epimers. This method was used to examine the stability of chlorophyll pigments of plant extracts obtained with various extraction methods including microwave-assisted extraction (MAE), supercritical fluid extraction (SFE), pressurized liquid extraction (PLE) and ultrasound-assisted extraction (UAE), with ethanol as solvent or modifier. The effect of storage was studied for both liquid and dried extracts. Irrespective of the extraction method, the evaporation of the extracts induced partial degradation of the chlorophyll pigments. It was found that liquid extracts could be stored at 4 °C for 3 weeks without a dramatic change in allomer forms of chlorophylls. However, during this storage period, epimerization appears to be important, leading to a significant decrease in the chlorophyll b native form. Graphical abstract.

The challenging management of Rift Valley Fever in humans: literature review of the clinical disease and algorithm proposal.

Rift Valley Fever (RVF) is an emerging zoonotic arbovirus with a complex cycle of transmission that makes difficult the prediction of its expansion. Recent outbreaks outside Africa have led to rediscover the human disease but it remains poorly known. The wide spectrum of acute and delayed manifestations with potential unfavorable outcome much complicate the management of suspected cases and prediction of morbidity and mortality during an outbreak. We reviewed literature data on bio-clinical characteristics and treatments of RVF human illness. We identified gaps in the field and provided a practical algorithm to assist clinicians in the cases assessment, determination of setting of care and prolonged follow-up.

Development of an analytical method for chlorophyll pigments separation by reversed-phase supercritical fluid chromatography.

Chlorophyll pigments give the green colour to plants, which is a quality attribute of food and vegetables. However, the chemical structure of native chlorophyll can change during varied processes (drying, freezing, extraction) applied to plants, which produce degradation compounds that could have a brown and unwanted colour. Systematic experiments have been conducted in supercritical fluid chromatography with a C18 stationary phase to understand and model the chromatographic behaviour of the compounds with respect to the nature of the modifier (MeOH, ACN, and MeOH/ACN 50/50) and its percentage, from 10% to 100%. Specific retention changes were observed, which provide numerous analytical conditions to achieve compound separation. The chromatographic profile of the extract containing native chlorophyll a, b and numerous phytylated chlorophyll derivatives (pheophytin a, a', b, b'; hydroxypheophytin a, a', b, b'; pyropheophytin and lactone derivatives) is strongly impacted by the nature of the modifier and, because of the complexity of the extract, the optimal conditions obtained are unusual for supercritical fluid chromatography. An original method development using an optimization criterion was discussed for the analyses of samples, leading to a fast analytical method with a very low backpressure and a flow rate gradient, but a simplest and rapid method is also suggested for samples displaying fewer derivatives.

Simultaneous extraction and determination of various pesticides in environmental waters.

A simple and rapid method was developed for the simultaneous analysis of nine different pesticides in water samples by gas chromatography with mass spectrometry. A number of parameters that may affect the recovery of pesticides, such as the type of solid-phase extraction cartridge, eluting solvent in single or combination and their volumes, and water pH value were investigated. It showed that three solid-phase extraction cartridges (Strata-X, Oasis HLB, and ENVI-18) produced the greatest recovery while ethyl acetate/dichloromethane/acetone (45:10:45, 12 mL) followed by dichloromethane (6 mL) was efficient in eluting target pesticides from solid-phase extraction cartridges. Different water pH values (4-9) did not show a significant effect on the pesticides recovery. The optimized method was verified by performing spiking experiments with a series of concentrations (0.002-10 µg/L) in waters, with good linearity, recovery, and reproducibility for most compounds. The limit of detection and limit of quantification of this optimized method were 0.01-2.01 and 0.02-6.71 ng/L, respectively, much lower than the European Union environmental quality standard for the pesticides (0.1 µg/L) in waters. The proposed method was further validated by participation in an interlaboratory trial. It was then subsequently applied to river waters from north-east Scotland, UK, for the determination of the target pesticides.