A Greener HPTLC Approach for the Determination of β-Carotene in Traditional and Ultrasound-Based Extracts of Different Fractions of Daucus carota (L.), Ipomea batatas (L.), and Commercial Formulation

Various analytical approaches for determining β-carotene in vegetable crops and commercial dosage forms have been documented. However, neither the qualitative nor quantitative environmental safety and greener aspects of the literature analytical methodologies of β-carotene analysis have been assessed. As a result, the goal of this research is to develop and validate a reversed-phase “high-performance thin-layer chromatography (HPTLC)” approach for determining β-carotene in traditional (TE) and ultrasound-assisted (UBE) extracts of different fractions of Daucus carota (L.), Ipomea batatas (L.), and commercial formulation. The greener mobile phase for β-carotene analysis was a ternary mixture of ethanol, cyclohexane, and ammonia (95:2.5:2.5, v v v−1). The detection of β-carotene was done at a wavelength of 459 nm. In the 25–1000 ng band−1 range, the greener reversed-phase HPTLC approach was linear. Other validation factors for β-carotene analysis, including as accuracy, precision, robustness, and sensitivity, were likewise dependable. The contents of β-carotene were found to be maximum in hexane: acetone (50:50%) fractions of TE and UBE of D. carota and I. batatas compared to their acetone and hexane fractions. The amount of β-carotene in hexane: acetone (50:50%) portions of TE of D. carota, I. batatas and commercial formulation A was estimated to be 10.32, 3.73, and 6.73 percent w w−1, respectively. However, the amount of β-carotene in hexane: acetone (50:50%) portions of UBE of D. carota, I. batatas and commercial formulation A was estimated to be 11.03, 4.43, and 6.89 percent w w−1, respectively. The greenness scale for the proposed HPTLC strategy was calculated as 0.81 using the “analytical GREEnness (AGREE)” method, indicating that the proposed HPTLC methodology has good greenness. The UBE approach for extracting β-carotene outperformed the TE procedure. These results indicated that the greener reversed-phase HPTLC approach can be utilized for the determination of β-carotene in different vegetable crops, plant-based phytopharmaceuticals, and commercial products. In addition, this approach is also safe and sustainable due to the utilization of a greener mobile phase compared to the toxic mobile phases utilized in literature analytical approaches of β-carotene estimation.

Rapid, Highly-Sensitive and Ecologically Greener Reversed-Phase/Normal-Phase HPTLC Technique with Univariate Calibration for the Determination of Trans-Resveratrol

The rapid, highly-sensitive and ecologically greener reversed-phase (RP)/normal-phase (NP) high-performance thin-layer chromatography (HPTLC) densitometric technique has been developed and validated for the determination of trans-resveratrol (TRV). The reversed-phase HPTLC-based analysis of TRV was performed using ethanol–water (65:35, v v−1) combination as the greener mobile phase, while, the normal-phase HPTLC-based estimation of TRV was performed using chloroform–methanol (85:15, v v−1) combination as the routine mobile phase. The TRV detection was carried out at 302 nm for RP/NP densitometric assay. The linearity was recorded as 10–1200 and 30–400 ng band−1 for RP and NP HPTLC techniques, respectively. The RP densitometric assay was observed as highly-sensitive, accurate, precise and robust for TRV detection in comparison with the NP densitometric assay. The contents of TRV in commercial formulation were recorded as 101.21% utilizing the RP densitometric assay, while, the contents of TRV in commercial formulation were found to be 91.64% utilizing the NP densitometric assay. The greener profile of RP/NP technique was obtained using the analytical GREEnness (AGREE) approach. The AGREE scales for RP and NP densitometric assays were estimated 0.75 and 0.48, respectively. The recorded AGREE scale for the RP densitometric assay indicated that this technique was highly green/the ecologically greener compared to the NP densitometric assay. After successful optimization of analytical conditions, validation parameters, AGREE scale and chromatography performance, the RP densitometric assay with univariate calibration was found to be better than the NP densitometric assay for the analysis of TRV.

PMMA bone cement containing long releasing silica-based chlorhexidine nanocarriers

Prosthetic joint infections (PJI) are still an extremely concerning eventuality after joint replacement surgery; growing antibiotic resistance is also limiting the prophylactic and treatment options. Chlorhexidine (a widely used topical non-antibiotic antimicrobial compound) coatings on silica nanoparticles capable of prolonged drug release have been successfully developed and characterised. Such nanocarriers were incorporated into commercial formulation PMMA bone cement (Cemex), without adversely affecting the mechanical performance. Moreover, the bone cement containing the developed nanocarriers showed superior antimicrobial activity against different bacterial species encountered in PJI, including clinical isolates already resistant to gentamicin. Cytocompatibility tests also showed non inferior performance of the bone cements containing chlorhexidine releasing silica nanocarriers to the equivalent commercial formulation.

A new culture medium based on genetic algorithms for Isochrysis galbana production relevant to hatcheries

The nutrient content of a commercial seawater culture medium for growing the microalga Isochrysis galbana was optimized using a stochastic strategy based on genetic algorithms. For this, 210 experiments spread over seven generations were carried out. This strategy reduced the number of assays by more than 90% compared to a factorial design involving the optimization of twelve nutrients simultaneously. The optimized medium outperformed the reference medium in all aspects. The genetic algorithm strategy achieved a polyunsaturated fatty acids (PUFAs) productivity of 7.8 mg L−1 day−1 in a continuous culture of I. galbana, corresponding to an increase of 15% compared to the commercial formulation. Carotenoids, on the other hand, increased by 50% d.w. In addition, PUFA yields were significantly improved, which allowed us to reduce the requirement of several nutrients, for instance, N (25%), Mo (20%), Mn (60%), Co (60%), and Cu (60%).

Effects of Prohydrojasmon on the Number of Infesting Herbivores and Biomass of Field-Grown Japanese Radish Plants

Prohydrojasmon (PDJ), an analog of jasmonic acid (JA), was found to induce direct and indirect defenses against herbivores in non-infested plants. To test whether PDJ can be used for pest control in crop production, we conducted experiments in pesticide-free Japanese radish fields from October 4 to December 12 in 2015. Twenty-four Japanese radish plants in three plots were treated with a 100 times-diluted commercial formulation (5%) of PDJ (treated plants), and 24 plants in three different plots were treated with water (control plants) until November 29 every week. Throughout the observation period, the number of aphids, leaf-mining fly larvae, vegetable weevils, and thrips was significantly lower on the treated plants than on the control plants. In contrast, the number of lepidopteran larvae was not significantly different between the treated and control plants throughout the study period. Parasitized aphids (mummies) were also observed in both plots. Poisson regression analyses showed that a significantly higher number of mummies was recorded on the treated plants as compared to that on the control plants when the number of aphids increased. This suggested that PDJ application to Japanese radish plants attracted more parasitoid wasps on the treated plants than on the control plants. We also identified eight terpenoids and methyl salicylate as the PDJ-induced plant volatiles in the headspace of the treated plants. Some of these volatiles might be responsible for attracting aphid-parasitoid wasps in the field. However, for other insect pests, we did not find any natural enemies. Interestingly, the genes of the JA and salicylic acid signaling pathways were differentially upregulated in the treated plants. We also observed that the PDJ treatments induced the expression of the genes related to glucosinolate biosynthesis and the subsequent isothiocyanate formation. Additionally, the weights of both the aboveground and belowground parts of the treated plants were significantly lower than those of the respective parts of the control plants. These results indicated that the treatment of Japanese radish plants with a 100 times-diluted commercial formulation of PDJ induced their direct and indirect defenses against several insect pest species to reduce their numbers, and negatively affected their biomass.

DIFFERENTIAL TOXICITY OF PYRETHROID AND ORGANOPHOSPHATE INSECTICIDES TO THE HONEY BEE, APIS MELLIFERA AND THE YELLOW FEVER MOSQUITO, AEDES AEGYPTI

Six insecticide active ingredients (AIs) and five commercial insecticide formulations were applied by topical application and onto filter paper strips to determine differential toxicity to Aedes aegypti (L.) and Apis mellifera (L.), and to evaluate their potential use in future insecticide resistance monitoring surveys. For topical application, 0.1 or 1 µl of the technical insecticide solution was applied to the Ae. aegypti and A. mellifera thorax, respectively. For insecticide-impregnated strips the insecticide amount varied, according with the commercial formulation. By topical application deltamethrin was the most toxic AI (LD 50 = 0.057 µg/g) to Ae. aegypti and prallethrin was least toxic (LD 50 = 19.42 µg/g). For A. mellifera, the most toxic AIs were deltamethrin (LD 50 = 0.013 µg/g) and bifenthrin (LD50 = 0.156 µg/g); and the least toxic was chlorpyrifos (LD 50 = 3.246 µg/g). When the insecticide-impregnated papers method was used, Mosquitomist Two (chlorpyrifos 24.6%) was the most toxic insecticide for Ae. aegypti (LC50 = 0.024 µg/cm2 ), and Aqualuer (permethrin 20.6%, PBO 20.6%) was least toxic (LC50 = 0.408 µg/cm2 ). For A. mellifera the most toxic commercial insecticide formulations were Talstar (bifenthrin 7.9%; LC50 = 0.288 µg/cm2 ) and Mosquitomist Two (LC50 = 0.299 µg/cm2 ), with no significant differences, and the least toxic commercial formulation was Deltagard (deltamethrin 2.0%; LC50 = 15.084 µg/cm2 ). By topical application, more than 28 times of chlorpyrifos was needed to obtain the same mortality in A. mellifera as in Ae. aegypti. When using the insecticide-impregnated paper method, more than 206 times of Deltagard was needed to obtain the same mortality in A. mellifera as in Ae. aegypti. Even though Mosquitomist Two was the most toxic insecticide for both insect species, the honey bees were >12 times more tolerant to this insecticide, compared with the mosquitoes.