Natural Product Extracts Research Articles

Natural Products Extraction of the Future—Sustainable Manufacturing Solutions for Societal Needs

The production of plant-based extracts is significantly influenced by traditional techniques and the natural variability of feedstock. For that reason, the discussion of innovative approaches to improve the manufacturing of established products and the development of new products within the regulatory framework is essential to adapt to shifting quality standards. This perspective of members of the DECHEMA/ProcessNet working group on plant-based extracts outlines extraction business models and the regulatory framework regarding the extraction of traditional herbal medicines as complex extracts. Consequently, modern approaches to innovative process design methods like QbD (Quality by Design) and quality control in the form of PAT (Process Analytical Technology) are necessary. Further, the benefit of standardized laboratory equipment combined with physico-chemical predictive process modelling and innovative modular, flexible batch or continuous manufacturing technologies which are fully automated by advanced process control methods are described. A significant reduction of the cost of goods, i.e., by a factor of 4–10, and decreased investments of about 1–5 mil. € show the potential for new products which are in line with market requirements.

Betulinic acid, derived from the desert lavender Hyptis emoryi, attenuates paclitaxel-, HIV-, and nerve injury-associated peripheral sensory neuropathy via block of N- and T-type calcium channels.

The Federal Pain Research Strategy recommended development of nonopioid analgesics as a top priority in its strategic plan to address the significant public health crisis and individual burden of chronic pain faced by >100 million Americans. Motivated by this challenge, a natural product extracts library was screened and identified a plant extract that targets activity of voltage-gated calcium channels. This profile is of interest as a potential treatment for neuropathic pain. The active extract derived from the desert lavender plant native to southwestern United States, when subjected to bioassay-guided fractionation, afforded 3 compounds identified as pentacyclic triterpenoids, betulinic acid (BA), oleanolic acid, and ursolic acid. Betulinic acid inhibited depolarization-evoked calcium influx in dorsal root ganglion (DRG) neurons predominantly through targeting low-voltage-gated (Cav3 or T-type) and CaV2.2 (N-type) calcium channels. Voltage-clamp electrophysiology experiments revealed a reduction of Ca, but not Na, currents in sensory neurons after BA exposure. Betulinic acid inhibited spontaneous excitatory postsynaptic currents and depolarization-evoked release of calcitonin gene-related peptide from lumbar spinal cord slices. Notably, BA did not engage human mu, delta, or kappa opioid receptors. Intrathecal administration of BA reversed mechanical allodynia in rat models of chemotherapy-induced peripheral neuropathy and HIV-associated peripheral sensory neuropathy as well as a mouse model of partial sciatic nerve ligation without effects on locomotion. The broad-spectrum biological and medicinal properties reported, including anti-HIV and anticancer activities of BA and its derivatives, position this plant-derived small molecule natural product as a potential nonopioid therapy for management of chronic pain.

Optimization of azo printing dye removal with oak leaves-nZVI/H2O2 system using statistically designed experiment

The paper investigates the potential use of nano zero-valent iron (nZVI) particles as a cheap, natural and effective catalyst in the Fenton-like system for the removal of water-based Magenta flexographic dye from aqueous solution. The functional nZVI was successfully synthesized within an environmentally friendly method, by using the natural product extract, oak leaves (OAK-nZVI). The process of Magenta removal was optimized using statistically designed experiment. The influence of four quantitative parameters on decolorization efficiency was investigated: initial dye concentration (20–180 mgL-1), OAK- nZVI dosage (0.75–60 mgL-1), H2O2 concentration (1–11 mM) and pH value (2–10). The results of statistical analysis show that OAK-nZVI dosage and pH significantly contribute to the decolorization efficiency, while the concentration of H2O2 has a significant influence on their impact. The optimization yielded highest removal efficiency of 91.95% under following conditions: initial dye concentration of 180 mgL−1, OAK-nZVI dosage of 60 mgL−1, H2O2 concentration of 11 mM and pH value 2. Optimization results were experimentally verified. Treatment of real effluent under optimized experimental conditions resulted within 84.06% Magenta removal after a reaction time of 60 min. The absorption spectra of Magenta dye clearly indicate the changes in the molecular structure in the form of the destruction of azo bond in the chromophore, leading to the decolorization of dye solution. Based on the GC–MS analysis, the mechanism for Fenton-like oxidation of Magenta was proposed.

CFD Simulation of Cross-Flow Filtration

The aim of the present work is to study the transfer processes in a cross-flow filtration cell, in order to determine the conditions for stable and efficient operation of a side-stream filtration module, integrated with a bioreactor. The current interest in membrane integrated bioreactors is connected with the pursuit of energy and cost efficiency in a wide area of industrial applications, including wastewater treatment, food industry, pharmaceutical industry, and fuel production. A numerical CFD model is employed, based on previous experience with experimental concentration of antioxidants, such as polyphenols and flavonoids from extracts of natural products by nanofiltration. The geometry under investigation is a 3D model of the experimental flat-sheet cell with tangential orientation of the feed inlet. The swirling turbulent flow in the feed channel is favourable for reducing the concentration polarization layer on the membrane surface and preventing fouling. The main factors, affecting the filtration process, are the shear stress distribution and the concentration profiles in the vicinity of the membrane surface. The CFD models of mass transfer in cross-flow nanofiltration are scarce and there are none for the reference experimental filtration cell. The present CFD simulation reveals the concentration distribution in the feed channel. It complements previous data for the flow pattern with new knowledge on the mass transfer there, directed to understanding and control of the concentration polarization phenomenon. The numerical study uses the tools of ANSYS Fluent R13, based on the finite volume method for solving the Reynolds-Averaged Navier-Stokes (RANS) equations. The obtained results are analysed in rapport with available experimental data.

Ionic Liquid Solutions as a Green Tool for the Extraction and Isolation of Natural Products.

In the past few years, the application of ionic liquids (ILs) had attracted more attention of the researchers. Many studies focused on extracting active components from traditional herbals using ILs as alternative solvents so as to address the issue caused by the traditional methods for extraction of natural products (NPs) with organic chemical reagents. Through the summary of reported research work, an overview was presented for the application of ILs or IL-based materials in the extraction of NPs, including flavonoids, alkaloids, terpenoids, phenylpropanoids and so on. Here, we mainly describe the application of ILs to rich the extraction of critical bioactive constituents that were reported possessing multiple therapeutic effects or pharmacological activities, from medicinal plants. This review could shed some light on the wide use of ILs in the field of natural products chemistry to further reduce the environmental damage caused by large quantity of organic chemical reagents.

Autophagy plays a protective role against apoptosis induced by toxicarioside N via the Akt/mTOR pathway in human gastric cancer SGC-7901 cells.

Toxicarioside N (Tox N), a natural product extract from Antiaris toxicaria, has been reported to induce apoptosis in human gastric cancer cells. However, the mechanism and actual role of autophagy in Tox N-induced apoptosis of human gastric cancer cells remains poorly understood. In the current study, we demonstrated that Tox N could induce autophagy by inhibiting the Akt/mTOR signaling pathway in SGC-7901 cells. Moreover, we found that the inhibition of autophagy by 3-methyladenine, an autophagy inhibitor, enhanced Tox N-induced apoptotic cell death. However, the stimulation of autophagy by rapamycin, an autophagy activator, remarkably suppressed Tox N-induced apoptosis, suggesting that autophagy plays a protective role in Tox N-induced apoptosis. Thus, the results from this study suggested that Tox N combination with an autophagy inhibitor might be a promising strategy to enhance the anticancer activity of Tox N for the treatment of human gastric cancer.

Microwave and enzyme co-assisted aqueous two-phase extraction of polyphenol and lutein from marigold (Tagetes erecta L.) flower

In this study, a novel method of simultaneous extraction and enrichment of total polyphenol (TP) and lutein from marigold flower was developed by microwave and enzyme co-assisted aqueous two-phase extraction (ATPE) (MEAATPE). In this study, ethanol/ammonium sulfate was chosen to construct aqueous two-phase system (ATPS) due to its fine partition and recycling behaviors. The optimal MEAATPE condition was obtained, that is, 28% (w/w), ethanol/20% (w/w) ammonium sulfate, 0.45 U/g enzyme concentration, enzymolysis time of 150 min, enzymolysis temperature of 45 °C, microwave time of 120 s, and microwave power of 270 W, by investigating the effects of ATPS composition and other factors on the yield of target products. Under this optimal condition, the yields of TP and lutein reached 84.61 and 7.32 mg/g with the corresponding recovery rates of 95.35% and 99.85%, respectively. Compared with enzyme-assisted ATPE, microwave-assisted ATPE, ATPE, and Soxhlet extraction (SE), the extraction yield of TP by MEAATPE was the highest, which is 71.14% higher than that of SE. Scanning electron microscopy analysis further showed an optative pretreatment effect of marigold flower powders. Thus, MEAATPE was an efficient, rapid, and environmental-friendly method used for the extraction of natural products.

Yeast-based screening of natural product extracts results in the identification of prion inhibitors from a marine sponge

ABSTRACTOne of the major medical challenges of the twenty-first century is the treatment of incurable and fatal neurodegenerative disorders caused by misfolded prion proteins. Since the discovery of these diseases a number of studies have been conducted to identify small molecules for their treatment, however to date no curative treatment is available. These studies can be highly expensive and time consuming, but more recent experimental approaches indicate a significant application for yeast prions in these studies. We therefore used yeast prions to optimize previous high-throughput methods for the cheaper, easier and more rapid screening of natural extracts. Through this approach we aimed to identify natural yeast-prion inhibitors that could be useful in the development of novel treatment strategies for neurodegenerative disorders. We screened 500 marine invertebrate extracts from temperate waters in Australia allowing the identification of yeast-prion inhibiting extracts. Through the bioassay-driven chemical investigation of an active Suberites sponge extract, a group of bromotyrosine derivatives were identified as potent yeast-prion inhibitors. This study outlines the importance of natural products and yeast prions as a first-stage screen for the identification of new chemically diverse and bioactive compounds.

Quick screening of true tyrosinase inhibitors from natural products using tyrosinase‐immobilized magnetic nanoparticles and a magnetic microplate

Tyrosinase inhibitors from natural products have applications in the pharmaceutical, food, and cosmetic industries because of the functions of tyrosinase in skin disorders and in the enzymatic browning of fruits. Current in vitro inhibitor screening assays are based on the inhibition of the oxidation of l‐3,4‐dihydroxyphenylalanine (l‐DOPA) mediated by a mushroom tyrosinase. However, in these assays, a tyrosinase inhibitor or an antioxidant could inhibit dopaquinone formation. In this study, we aimed to eliminate this ambiguity by using a microplate assay integrating tyrosinase‐immobilized magnetic nanoparticles (TYR‐MNPs) and a homemade magnetic microplate for high‐throughput screening. After incubating extracts of natural products with TYR‐MNPs, the magnetic nanoparticles are attracted to the bottoms of wells, the extracts are rinsed, and TYR‐MNPs react with l‐DOPA. This method can be used to screen compounds that interact with the active sites of the enzyme, or copper chelators that bind more strongly than tyrosinase to copper ions, distinguishing them from antioxidants or tyrosinase substrates. Integration with the homemade magnetic microplate enables high‐throughput inhibitor screening. Aloe vera flowers are crop by‐products, and litchi flowers fall after the blossom. Our work demonstrated that these flowers have tyrosinase inhibitory effects, thus increasing their value.

Out of the sea and into the factory

When Pierre Lemieux and his team at Acasti Pharma went looking for a pharmaceutical manufacturing partner, they had a surprising amount of trouble finding one. The reason was the type of drug they are developing. The active ingredient in Acasti’s lead product, which lowers triglyceride levels in people at risk of cardiovascular disease, is an extract from krill oil, and not everyone wants to work with krill oil. For starters, natural product extraction is an art not widely practiced among drug outsourcing firms. In addition, krill are shrimplike crustaceans, and some potential partners were concerned about contaminating their facilities with shellfish allergens. On top of that, extracting the active ingredient involves acetone, a flammable solvent some companies weren’t prepared to handle. A krill oil plant operated by Acasti’s then parent company exploded in 2012, killing three people. But Acasti kept looking. It was having success in early trials of its

Toward a Distinct and Quantitative Validation Method for Predictive Process Modelling—On the Example of Solid-Liquid Extraction Processes of Complex Plant Extracts

Physico-chemical modelling and predictive simulation are becoming key for modern process engineering. Rigorous models rely on the separation of different effects (e.g., fluid dynamics, kinetics, mass transfer) by distinct experimental parameter determination on lab-scale. The equations allow the transfer of the lab-scale data to any desired scale, if characteristic numbers like e.g., Reynolds, Péclet, Sherwood, Schmidt remain constant and fluid-dynamics of both scales are known and can be described by the model. A useful model has to be accurate and therefore match the experimental data at different scales and combinations of process and operating parameters. Besides accuracy as one quality attribute for the modelling depth, model precision also has to be evaluated. Model precision is considered as the combination of modelling depth and the influence of experimental errors in model parameter determination on the simulation results. A model is considered appropriate if the deviation of the simulation results is in the same order of magnitude as the reproducibility of the experimental data to be substituted by the simulation. Especially in natural product extraction, the accuracy of the modelling approach can be shown through various studies including different feedstocks and scales, as well as process and operating parameters. Therefore, a statistics-based quantitative method for the assessment of model precision is derived and discussed in detail in this paper to complete the process engineering toolbox. Therefore a systematic workflow including decision criteria is provided.

NCI Program for Natural Product Discovery: A Publicly-Accessible Library of Natural Product Fractions for High-Throughput Screening.

The US National Cancer Institute's (NCI) Natural Product Repository is one of the world's largest, most diverse collections of natural products containing over 230,000 unique extracts derived from plant, marine, and microbial organisms that have been collected from biodiverse regions throughout the world. Importantly, this national resource is available to the research community for the screening of extracts and the isolation of bioactive natural products. However, despite the success of natural products in drug discovery, compatibility issues that make extracts challenging for liquid handling systems, extended timelines that complicate natural product-based drug discovery efforts and the presence of pan-assay interfering compounds have reduced enthusiasm for the high-throughput screening (HTS) of crude natural product extract libraries in targeted assay systems. To address these limitations, the NCI Program for Natural Product Discovery (NPNPD), a newly launched, national program to advance natural product discovery technologies and facilitate the discovery of structurally defined, validated lead molecules ready for translation will create a prefractionated library from over 125,000 natural product extracts with the aim of producing a publicly-accessible, HTS-amenable library of >1,000,000 fractions. This library, representing perhaps the largest accumulation of natural-product based fractions in the world, will be made available free of charge in 384-well plates for screening against all disease states in an effort to reinvigorate natural product-based drug discovery.

Targeting Protein Quality Control Mechanisms by Natural Products to Promote Healthy Ageing.

Organismal ageing is associated with increased chance of morbidity or mortality and it is driven by diverse molecular pathways that are affected by both environmental and genetic factors. The progression of ageing correlates with the gradual accumulation of stressors and damaged biomolecules due to the time-dependent decline of stress resistance and functional capacity, which eventually compromise cellular homeodynamics. As protein machines carry out the majority of cellular functions, proteome quality control is critical for cellular functionality and is carried out through the curating activity of the proteostasis network (PN). Key components of the PN are the two main degradation machineries, namely the ubiquitin-proteasome and autophagy-lysosome pathways along with several stress-responsive pathways, such as that of nuclear factor erythroid 2-related factor 2 (Nrf2), which mobilises cytoprotective genomic responses against oxidative and/or xenobiotic damage. Reportedly, genetic or dietary interventions that activate components of the PN delay ageing in evolutionarily diverse organisms. Natural products (extracts or pure compounds) represent an extraordinary inventory of highly diverse structural scaffolds that offer promising activities towards meeting the challenge of increasing healthspan and/or delaying ageing (e.g., spermidine, quercetin or sulforaphane). Herein, we review those natural compounds that have been found to activate proteostatic and/or anti-stress cellular responses and hence have the potential to delay cellular senescence and/or in vivo ageing.

Supercritical Fluid Extraction of High-value Natural Products from Buttermilk Analyzed by a Dynamic Extraction Model

本研究では，経口物質に対して安心安全な製造法である超臨界流体を用いて，乳製品副産物に含まれる機能性成分であるphosphatidylcholineとsphingomyelinの効率的分画を目的としてこれら成分の溶解度推定を検討した．まず既往の実験データのある，バターオイルからのtriacylglycerolにおける超臨界CO2中の溶解度と比較を行った結果，本研究で推定した溶解度とほぼ一致することが確認された．次に，PCおよびSMの溶解度の溶媒密度およびエタノール濃度依存性の解析を行った結果，いずれの条件においても実験的な溶解度を推定することができた．PCおよびSMは高価であり標準試薬が入手しにくい状況でこれまで超臨界流体中の溶解度データも報告されていなかったが，本研究で行った抽出モデルによる解析により溶解度を実験的に推定することができた．

The order of calculation and payment of tax on profit of commercial banks in the innovation economy

In the conditions of innovative economy of the tightening of the requirements of the Central Bank of the Russian Federation and the crisis of the banking system, in the conditions of a severe shortage of funds in the Russian Federation, taxes and fees levied on credit institutions are one of the main revenue sources of the state budget after the extraction, processing, transportation, warehousing and sale of natural resources-oil, gas and related petroleum products. In practice, the taxation of profits of credit institutions is under the close attention of the state. So, PJSC "Sberbank of Russia" in 2016, took fifth place among the largest Russian taxpayers, losing the leadership of the JSC "NK Rosneft", PJSC "Gazprom", PJSC "LUKOIL" and JSC "Surgutneftegas". At the same time in 2015 PJSC "Sberbank of Russia" has taken only the tenth place. A particularly urgent task in modern conditions is the reduction of the strongly expressed fiscal orientation of taxes and fees and the increase of their motivating role, as well as the strengthening of the legal regulation of fees and taxes as a complex part of the legislation in the field of taxation of Russia as a whole. The article describes taxpayers, the object of taxation on income tax of a commercial Bank. The detailed calculation of the tax base of income tax of PJSC "Sberbank of Russia" by year is presented. The table of calculation of the Bank's income is provided, by results of the analysis it is revealed that the greatest share in structure of the income is occupied by interest income all three years about 90%, however dynamics of development of banking sector and global tendencies dictate growth of Commission income in structure of the General income. Distinct dynamics can be traced clearly. The calculation of expenses for three years of PJSC “Sberbank of Russia” is also presented, the analysis is carried out, conclusions are formulated. According to the results of the work the conclusion about the need for further development of the system of taxation of commercial banks in obligatory correlation with the strategy and objectives of development of other sectors of the national economy of Russia.

Establishment of an HPLC method for testing acetylcholinesterase inhibitory activity and compared with traditional spectrophotometry

Acetylcholinesterase (AChE) inhibitory activity test is one of the important areas of natural product research, which is traditionally tested by spectrophotometry. In this work, a simple high-performance liquid chromatography (HPLC) method was developed for testing AChE inhibitory activity in several natural products. Using galanthamine hydrobromide and huperzine A as positive controls, it was found that the optimal conditions for sample preparation are 7.0–8.0 for pH, 30 min for reaction time, 37 °C for reaction temperature, 1.1 × 10−3 U mL−1 for concentration of AChE, and 0.625 mM for acetylthiocholine iodide in the final reaction mixture with total volume 2 mL. The Diamonsil C18 column was used and the chromatographic conditions were mobile phase methanol–water–triethylamine (40:60:0.05, v/v/v), flow rate 1.0 mL min−1, injection volume 5 μL, run time 5 min, column temperature 37 °C, and wavelength 405 nm. The AChE inhibitory activity of anthocyanin with concentration of 10, 20, and 50 μg mL−1, blueberry extract and purple potato extract with concentration of 20, 100, and 500 μg mL−1 were further determined under this conditions. The results showed the inhibition ratios obtained from spectrophotometry were nearly adjacent and approximately lower than 10% and the values of absorbance mostly exceed 0.8, but the absorbance should ensure in the range of 0.15–0.8 to guarantee smaller relative error for spectrophotometry. While the inhibition ratios gained from HPLC method were higher than 20% and had greater difference, they had better comparability to the different concentrations of anthocyanin and its extracts. It is revealed that the HPLC method can be used for the evaluation of the AChE inhibitory activity in natural product extracts with deep color or not.

A solid-state microwave method to disrupt biomass microstructure for natural product extraction

This paper introduces an innovative solid-state microwave disruption (SSMD) method for the extraction of bioactive materials from plants. It is an alternative method that avoids the use of solvents for plant biomass sample pretreatment unlike those in the conventional microwave methods. SSMD method is preferred since the heating and the vaporization of the intracellular moisture within the plant biomass is able to create sufficient pressure to disrupt the biomass microstructure. In this work, the superiority of the SSMD method was validated using the bioactive extraction from Orthosiphon stamineus (Java tea) leaves. First, the degree of disruption of SSMD method under the effects of microwave power and heating time were investigated at various up-scaled conditions based on the structural analysis of the treated sample, and the extraction yields of the bioactive compounds after soaking and elution with a suitable solvent at room conditions. The results show that the SSMD-elution method gave a better extraction yield (4.65±0.08mg/g), which is comparable to that of microwave-assisted extraction (4.45±0.03mg/g) and ultrasonic-assisted extraction (4.18±0.02mg/g). In addition, this method has great operational simplicity and flexibility in terms of operating parameters, types of solvent and solvent consumption.

CHEMICAL CONSTITUENTS FROM ETHYL ACETATE EXTRACT OF SCOPARIA DULCIS LINN.

This research focuses on the isolation and characterization of eight natural compounds fromthe ethyl acetate extract of Scoparia dulcis L. collected in An Giang province, Vietnam. Sixflavonoid compounds: apigenin (1), luteolin (2), baicalein (3), oroxylin A (4), oroxylin A 7-O-β-D-glucoside (5) and vitexin (6) along with two nitrogen-containing compounds: 2,4-dihydroxy-1,4-benzoxazin-3-one (7) and uracil (8) were successfully isolated via the methods of naturalproduct extraction. The structures of these compounds were elucidated base on 1D-NMR and2D-NMR spectral data as well as MS and compared with those in the authentic reports. Thisstudy contributed new results to the phytochemical examination of Scoparia dulcis Linn.growing in Vietnam with the presence of the first isolated compounds as (3), (4), (5), (7) and (8).

Crystalline‐Sponge‐Based Structural Analysis of Crude Natural Product Extracts

AbstractCharacterization of complex natural product mixtures to the absolute structural level of their components often requires significant amounts of starting materials and lengthy purification process, followed by arduous structure elucidation efforts. The crystalline sponge (CS) method has demonstrated utility in the absolute structure elucidation of isolated organic compounds at miniscule quantities compared to conventional methods. In this work, we developed a new CS‐based workflow that greatly expedites the in‐depth structural analysis of crude natural product extracts. Using a crude extract of the red alga Laurencia pacifica, we showed that CS affinity screening prior to compound isolation enables prioritization of analytes present in the extract, and we subsequently resolved the molecular structures of six sesquiterpenes with stereochemical clarity from around 10 mg crude extract. This study demonstrates a new chemotyping workflow that can greatly accelerate natural product discovery from complex samples.

Crystalline-Sponge-Based Structural Analysis of Crude Natural Product Extracts.

Characterization of complex natural product mixtures to the absolute structural level of their components often requires significant amounts of starting materials and lengthy purification process, followed by arduous structure elucidation efforts. The crystalline sponge (CS) method has demonstrated utility in the absolute structure elucidation of isolated organic compounds at miniscule quantities compared to conventional methods. In this work, we developed a new CS-based workflow that greatly expedites the in-depth structural analysis of crude natural product extracts. Using a crude extract of the red alga Laurencia pacifica, we showed that CS affinity screening prior to compound isolation enables prioritization of analytes present in the extract, and we subsequently resolved the molecular structures of six sesquiterpenes with stereochemical clarity from around 10 mg crude extract. This study demonstrates a new chemotyping workflow that can greatly accelerate natural product discovery from complex samples.