María Fernanda Silva

Metal content monitoring in Hypericum perforatum pharmaceutical derivatives by atomic absorption and emission spectrometry

Metals have been investigated in different plant materials in order to establish their normal concentration range and consider their role in plants as part of human medicinal treatment. Metal monitoring as a pattern recognition method is a promising tool in the characterization and/or standardization of phytomedicines. In the present work measurable amounts of Ca, Cu, K, Li, Mg, Mn, Na, Ni, and Zn were detected in phytopharmaceutical derivatives of Hypericum perforatum by atomic techniques. Atomic methodologies like flame atomic absorption spectrometry (FAAS) and electrothermal atomic absorption spectrometry (ETAAS) allow reliable determination of mineral content in pharmaceutical quality control of medicinal plants. Additionally, capillary electrophoresis (CE) patterns of characteristic components (fingerprints) have been performed for the search of adulterants in phytopharmaceutical products.

Simultaneous determination of cloramphenicol, salicylic acid and resorcinol by capillary zone electrophoresis and its application to pharmaceutical dosage forms

This study demonstrates the separation of active ingredients in acne formulations (salicylic acid, cloramphenicol and resorcinol in presence of azulene) by capillary zone electrophoresis. Factors affecting their separations were the buffer pH and concentration, applied voltage, sample preparation, and presence of additives. Optimun results were obtained with a 50 mM sodium tetraborate-50 mM sodium phosphate, pH 9.0. The carrier electrolyte gave baseline separation with good resolution, short migration times (<6 min), great reproducibility and accuracy. Calibration plots were linear over at least three orders of magnitude of analyte concentrations, the lower limits of detection being within the range 0.39-1.25 mug ml(-1). The procedure was fast and reliable and commercial pharmaceuticals could be analysed without prior sample clean-up procedure.

Nectar and Flower Traits of Different Onion Male Sterile Lines Related to Pollination Efficiency and Seed Yield of F1 Hybrids

ABSTRACT Honey bees are the main pollinators of onion crops for seed production, but owing to low attractiveness of flowers, pollination is often inadequate. Pollination problems result in low seed yields. This problem is accentuated when male sterile lines (MSL) are used to produce hybrid onion seeds. In this study, the effect of floral attributes and nectar composition on the preference of honey bees of four MSLs and one onion open pollinated cultivar were assessed. The chemical composition of nectar was described through the analysis of sugars, trace elements, volatile organic compounds, and phenol compounds. The samples studied showed qualitative and quantitative differences in the analyzed traits of flowers and nectar among the different lines. Furthermore, field observations showed a great difference on the number of bee visits and seed yield among the onion lines analyzed. For the first time, this study demonstrates that there are marked differences in the chemical composition of nectar and floral morphology between open pollinated and MSLs and also within MSLs. In addition, these differences were correlated with the number of visits and seed yield. Therefore, it would be possible to select indirectly the most promising productive MSL using simple determinations of chemical compounds or floral morphological characters.

Determination of seleno-amino acids bound to proteins in extra virgin olive oils.

An analytical method has been developed to determine seleno-amino acids in proteins extracted from extra virgin olive oils (EVOOs). Different aqueous/organic solvents were tested to isolate proteins, an acetone:n-hexane combination being the best protein precipitant. In a first dimension chromatography, extracted proteins were analysed by size exclusion chromatography (SEC) coupled to inductively coupled plasma mass spectrometry (ICP-MS) to identify S and Se associations as proteins marker. Two fractions of 66 kDa (A) and 443 kDa (B) were identified. These fractions were submitted to microwave-assisted acid hydrolysis (MAAH) to release seleno-amino acids. In a second dimension chromatography seleno-amino acids were determined by reversed-phase chromatography (RPC) coupled to ICP-MS. Seleno-methylselenocysteine was determined with values ranging from 1.03-2.03±0.2 μg kg(-1) and selenocysteine at a concentration of 1.47±0.1 μg kg(-1). Variations of protein and seleno-amino acid concentrations were observed between EVOO varieties, contributing to EVOO cultivar differentiation.

Preconcentration of seleno-amino acids on a XAD resin and determination in regional olive oils by SPE UPLC–ESI-MS/MS

This study describes a method for seleno-amino acids determination in Argentinean olive oils. Preliminary total selenium determination in olive oils probed low concentrations (62.8±1.6 to 117.4±3.0 μg/kg) and the necessity of implementing a preconcentration method. To this end a XAD® resin was employed as sorbent for selenomethionine (Se-Met), selenomethylselenocysteine (Se-MetSeCys), and selenocysteine (Se-Cys) preconcentration. Determinations were performed by UPLC-ESI-MS/MS. Recoveries were between 84% and 97% for the seleno-amino acids studied, reaching a detection limit of 0.09 μg/kg, a precision of 10% (RSD, n=6), and an enhancement factor of 60-fold (6 for the extraction system and 10 for the preconcentration approach). The only detected Se species in the olive oils was Se-MetSeCys in concentrations ranging from 2.0 to 8.3 μg/kg.

Determination of alkaloids in onion nectar by micellar electrokinetic chromatography.

Nectar is the most important floral reward offered by plants to insects. Minor components such as alkaloid compounds in nectar affect bee foraging, with great influence in seed production. CE is an advantageous tool for the analysis of unexplored samples such as onion nectar due to the limited amounts of samples. Considering the importance of these compounds, a simultaneous determination of nicotine, theophylline, theobromine, caffeine, harmaline, piperine in onion nectar by MEKC‐UV is herein reported. The extraction of alkaloid compounds in nectar was performed by SPE using a homemade miniaturized column (C18). Effects of several important factors affecting extraction efficiency as well as electrophoretic performance were investigated to acquire optimum conditions. Under the proposed conditions, the analytes can be separated within 15 min in a 50 cm effective length capillary (75 μm id) at a separation voltage of 20 kV in 20 mmol/L sodium tretraborate, 100 mmol/L SDS. The amount of sample requirement was reduced up to 2000 times, when compared to traditional methods, reaching limits of detection as low as 0.0153 ng/L. For the first time, this study demonstrates that there are marked qualitative and quantitative differences in nectar alkaloids between open pollinated and male sterile lines (MSLs) and also within MSLs.

Onion Hybrid Seed Production: Relation with Nectar Composition and Flower Traits

Abstract Onion (Allium cepa L.) is one of the main vegetable crops. Pollinators are required for onion seed production, being honeybees the most used. Around the world, two types of onion varieties are grown: open pollinated (OP) and hybrids. Hybrids offer numerous advantages to growers, but usually have lower seed yields than OP cultivars, which in many cases compromise the success of new hybrids. As pollination is critical for seed set, understanding the role of floral rewards and attractants to pollinator species is the key to improve crop seed yield. In this study, the correlation of nectar-analyzed compounds, floral traits, and seed yield under open field conditions in two experimental sites was determined. Nectar composition was described through the analysis of sugars, phenol, and alkaloid compounds. Length and width of the style and tepals of the flowers were measured to describe floral traits. Floral and nectar traits showed differences among the studied lines. For nectar traits, we found a significant influence of the environment where plants were cultivated. Nonetheless, flower traits were not influenced by the experimental sites.The OP and the male-sterile lines (MSLs) showed differences in nectar chemical composition and floral traits. In addition, there were differences between and within MSLs, some of which were correlated with seed yield, bringing the opportunity to select the most productive MSL, using simple determinations of morphological characters like the length of the style or tepals size.