A Descriptive Chemical Composition of Concentrated Bud Macerates through an Optimized SPE-HPLC-UV-MS2 Method—Application to Alnus glutinosa, Ribes nigrum, Rosa canina, Rosmarinus officinalis and Tilia tomentosa

Concentrated bud macerates (CBMs) are obtained from meristematic tissues such as buds and young shoots by maceration in a solvent composed of glycerin, water and ethanol (1/1/1/, v/v). Their traditional utilization in gemmotherapy has gained interest in the past years, and the knowledge of their chemical characterization can provide commercial arguments, particularly to secure their quality control. Therefore, an optimized method for phytochemical analysis including glycerol removal by a preliminary solid phase extraction (SPE) followed by compound identification using high performance liquid chromatography coupled with ultra-violet and tandem mass detectors (HPLC-UV-MS2) was developed. This method was applied on 5 CBMs obtained from Alnus glutinosa, Ribesnigrum, Rosmarinus officinalis, Rosa canina and Tilia tomentosa in order to determinate their chemical composition. Their antioxidant effects were also investigated by radical scavenging activity assays (DPPH and ORAC). Glycerol removal improved the resolution of HPLC chemical profiles and allowed us to perform TLC antioxidant screening. Our approach permitted the identification of 57 compounds distributed in eight major classes, three of them being common to all macerates including nucleosides, phenolic acids and glycosylated flavonoids. Quantification of the later class as a rutin equivalent (RE) showed a great disparity between Rosa canina macerate (809 mg RE/L), and the other ones (from 175 to 470 mg RE/L). DPPH and ORAC assays confirmed the great activity of Rosa canina (4857 and 6479 μmol TE/g of dry matter, respectively). Finally, phytochemical and antioxidant analysis of CBMs strengthened their phytomedicinal interest in the gemmotherapy field.

Influence of Gender and Age of Brown Seaweed (Fucus vesiculosus) on Biochemical Activities of Its Aqueous Extracts

Fucus vesiculosus L. is a common coastal brown seaweed associated with various benefits to human health due to its phenolic content and nutrients and is used as food through different methods of consumption. This study aims to evaluate the influence of the seaweed’s gender and growth stage on different types of biological activities as well as its chemical constitution and elements present. Akin to food preparation, aqueous extracts of the seaweed were prepared at 25 °C (salad) and 100 °C (soup). Biological activities were determined by measuring total phenol content (TPC), antioxidant activity and inhibition of acetylcholinesterase (AChE). Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS/MS) was used for compound identification, and elemental analysis was carried out by using Total Reflection X-ray Fluorescence Spectrometry (TXRF). Older females and males had higher TPC compared to the new ones at 100 °C. Antioxidant activity depended on the extraction temperature but was higher for the youngest male at 100 °C. AChE inhibitory activity was higher for older males at 25 °C, but at 100 °C it was higher for older females. Primary metabolites and various phloroglucinol were the main compounds identified. Additionally, since this seaweed is often harvested in estuarine systems with high anthropogenic impacts, its safety was evaluated through the evaluation of the sample’s metal content. The heavy metals detected are within the limits established by various regulating entities, pointing to a safe food source.

POTENTIAL ANTIDIABETIC ACTIVITIES OF FRACTIONS FROM PURIFIED EXTRACT OF LAWSONIA INERMIS LEAVES IN ALLOXAN–INDUCED DIABETIC MICE

Objective: This research was conducted to determine the potential antidiabetic activity fractions of purified extract Lawsonia inermis leaves in mice (Mus musculus) and identification of the compound. Methods: The method included maceration, purification using ethanol and distilled water was followed by liquid-liquid extraction using ethyl acetate and magnesium sulfate as drying agents. Furthermore, the extract was analyzed using thin layer chromatography (TLC) for testing the purified extract. Fractionation using vacuum liquid chromatography, antidiabetic activity test of fractions at dose 100 mg/kgBW with alloxan induced and compound identification by Liquid Chromatography-Mass Spectrometry (LC-MS/MS) using HPLC connected to a Q-TOF spectrometer equipped with an ESI source, with Phenomenon column C8, and methanol with 0.3% formic acid as solvent. Results: The results showed that from the purification step of L. inermis leaves by vacuum liquid chromatography method, 7 fractions were obtained, i.e. A-G fractions. While the antidiabetic activity of fractions shown by decreasing blood sugar level in mice on the 15th day were 64, 75, 73, 73, 57, 45 and 67%, respectively. The identified compounds from each fraction were the ester groups namely 12-hydroxy-methyl abietate, 9,12-octadecadienoic acid (Z,Z)-(2,2-dimethyl-1,3-dioxolan-4-yl)methyl ester, dehydromorroniaglycone, and (E)-hexadecyl-ferulate; the steroid group namely siraitic acid E; phenylpropanoid groups namely umbelliferone and bletilol C, and the alkaloid groups namely moupinamide and valine. Conclusion: L. inermis leaves had activity in lowering blood sugar levels. LC-MS/MS analysis revealed the presence of ester groups, steroid groups, phenylpropanoid groups and alkaloid groups. The presence of these compounds mostly contribute to antidiabetic activity.

Mass Spectrometry Coupled with Chromatography toward Separation and Identification of Organic Mixtures

Mass spectrometers can provide information about molecular composition and chemical structure. However, with complex mixtures, superpositions and even suppression of signals may occur. On the other hand, Chromatography is an ideal technique for separating complexes but is often insufficient for compound identification. Hence, coupling both techniques in order to eliminate the limitations of each technique makes perfect sense. In this contribution, a brief description of mass spectrometry coupled with chromatography in the gas and liquid phase will be discussed to explain the advantages of coupling the two methods. The ionization techniques are also reported and followed by application areas of these techniques. Finally, the recording and treatment of the results are reviewed.

Paper Chromatographic Separation of Plant Pigments a New Approach by using Akshya-Swagatika Solvent and Mobile Chromatogram Detection System (MCDS)

Chromatography is a term that refers to a group of laboratory techniques for separating mixtures. Chromatography works on the premise of solute partitioning between two phases or solvents. The technique of paper chromatography is commonly used to separate plant pigments based on their molecular weight. Plant pigments include chlorophyll-b, chlorophyll-a, carotenoid, and xanthophyll, which all have various molecular weights, colours, and absorption maxima. In this study, an attempt was made to see how a new solvent (Akshya-Swagatika solvent) can be used to separate plant pigments using paper chromatography, as well as a new detection method developed by us known as Mobile chromatogram detection system (MCDS) that can be used for compound identification and photographing. As shown in the table and photograph, pigments are separated by molecular weight and band colour, with carotenoid having the highest RF value and chlorophyll-b having the lowest. It was obvious that the Akshya-Swagatika solution could be used to separate plant pigments in paper chromatography. Photograph taken with a mobile chromatogram detection system that is much clearer than normal one. In developing countries, both solvent and detection systems are useful in explaining paper chromatography in a cost-effective manner. The MCDS detection system is cost-effective fast method which replaces traditional sophisticated detection procedures.

A Fast and Automated Strategy for the Identification and Risk Assessment of Unknown Substances (IAS/NIAS) in Plastic Food Contact Materials by GC-Q-Orbitrap HRMS: Recycled LDPE as a Proof-of-Concept

A fast and automated approach has been developed for the tentative identification and risk assessment of unknown substances in plastic food contact materials (FCM) by GC-Q-Orbitrap HRMS. The proposed approach combines GC-HRMS full scan data acquisition coupled to Compound Discoverer™ 3.2 software for automated data processing and compound identification. To perform the tentative identification of the detected features, a restrictive set of identification criteria was used, including matching with the NIST Mass Spectral Library, exact mass of annotated fragments, and retention index calculation. After the tentative identification, a risk assessment of the identified substances was performed by using the threshold of toxicological concern (TTC) approach. This strategy has been applied to recycled low-density polyethylene (LDPE), which could be used as FCM, as a proof-of-concept demonstration. In the analyzed sample, 374 features were detected, of which 83 were tentatively identified after examination of the identification criteria. Most of these were additives, such as plasticizers, used in a wide variety of plastic applications, oligomers of LDPE, and substances with chemical, industrial, or cosmetic applications. The risk assessment was performed and, according to the TTC approach, the obtained results showed that there was no risk associated with the release of the identified substances. However, complementary studies related to the toxicity of the unidentified substances and the potential mixture toxicity (cocktail effects) should be conducted in parallel using bioassays.

Development of the method for determining Methandienone in Toxicology and doping-analysis

Anabolic-androgenic steroids are ones of the most frequently detected drugs in amateur and professional sports. Doping control laboratories have developed numerous assays enabling the determination of administered drugs. However, it is relevant today to improve detection methods that are accurate, fast and require small amounts of reagents. The identification and quantification of methandienone from body fluids has been insufficiently developed. The gas chromatography-mass spectrometry (GC-MS) method for the determination of methandienone traces in body fluids was developed and validated in the toxicology laboratory in order to control a non-medical use of an anabolic steroid-methandienone. Extracting procedures were optimized in order to obtain entire amount of the drug and methandienone and its metabolites were determined by sensitive and specific reactions for the compound identification, as well as the system for carrying out the TLC method have been proposed, and conditions for identifying a test sample from the biomaterial by IR spectrophotometry have been selected. The GC-MS method is found to be simple, fast, sensitive, and the strategy proposed can be effortlessly and advantageously applied for the routine analysis of biological fluids for determining methandienone and its metabolites at a level less than 30 ng in toxicology and doping-analysis.