Chlorophyll Extraction Research Articles

Determination of bioactive properties of protein and pigments obtained from Spirulina platensis

Microalgae have a great potential for usage in functional foods or ingredients. The amount of the pigments obtained with different solvents from Spirulina platensis and their antioxidant activities, Angiotensin Converting Enzyme (ACE) inhibition of S. platensis and dried phycocyanin powder were investigated. The highest Chlorophyll-a and phycocyanin pigment contents were 9.26 mg chlorophyll-a/g dry spirulina and 78.57 mg phycocyanin/g dry spirulina using sequentially extraction, respectively. It was found that Chlorophyll extracts have higher antioxidant activity (14.62–16.19 mg FeSO4·7H2O/g dry spirulina) than phycocyanin extracts (10.87–12.34 mg FeSO4·7H2O/g dry spirulina). Single extraction was preferred due to increased pigment concentration and antioxidant activity. The percent ACE inhibition values of 10% S. platensis powder and phycocyanin powder suspensions were found as 92.40% and 89.50%, respectively. The necessary protein concentration of S. platensis and phycocyanin powder samples to inhibit 50% of the ACE activity (IC50 values) were 2.93 and 3.96 mg protein/ml, respectively. It was concluded that S. platensis suspension could be more effective than the pure phycocyanin on ACE inhibition. Practical applications Microalgae itself can be considered as functional food or food ingredient because of the bioactive contents such as polyunsaturated fatty acids, sterols, pigments, proteins and enzymes, vitamins, and other bioactive substances. In this study, the quantities and the antioxidant activities of phycocyanin and chlorophyll-a pigments obtained from Spirulina platensis by sequential extraction of methanol and aqueous sodium nitrate solutions were determined. Also, protein contents and ACE inhibition of residual protein rich fraction were determined after the pigment extraction. The results showed that pigment fractions and residual protein content after pigment extraction have antioxidative and antihypertensive health promoting potential.

Effects of solar drying operation equipped with a finned and double-pass heat collector on energy utilization, essential oil extraction and bio-active compounds of peppermint (Mentha Piperita L.)

Three methods of shade (MI), open sun (MII) and new designed of solar (MIII) drying were used for peppermint dehydration. The air velocity (u) and temperature (Td ) of MI, MII and MIII were 0.5, 0.5, and 1.5 m/s and 32, 36, and 48 °C, respectively. The energy utilization efficiency, Td /u ratio and drying rate of MIII reached 85%, 48 °C.s.m−1 and 12.5 gH2O/min, respectively, which were higher than those in MI and MII. The inhibition power of dried peppermints against 6-log colony forming unit/ml of Staphylococcus aureus (Sa) were also evaluated. While the chlorophyll and essential oil extraction efficiency of MIII-product were 30% and 60% respectively higher than those in MII-product, its total phenolic contents and antioxidants activities were 25% and 26% more than those in MI-product. The essential oil of MIII-product had significantly (p < 0.05) higher inhibiting compounds (mainly Menthone) against pathogenic gram-positive Sa compared with those in MI and MII-products.

Poly(styrene-co-maleic acid)-mediated isolation of supramolecular membrane protein complexes from plant thylakoids

Derivatives of poly(styrene-co-maleic acid) (pSMA), have recently emerged as effective reagents for extracting membrane protein complexes from biological membranes. Despite recent progress in using SMAs to study artificial and bacterial membranes, very few reports have addressed their use in studying the highly abundant and well characterized thylakoid membranes. Recently, we tested the ability of twelve commercially available SMA copolymers with different physicochemical properties to extract membrane protein complexes (MPCs) from spinach thylakoid membrane. Based on the efficacy of both protein and chlorophyll extraction, we have found five highly efficient SMA copolymers: SMA® 1440, XIRAN® 25010, XIRAN® 30010, SMA® 17352, and SMA® PRO 10235, that show promise in extracting MPCs from chloroplast thylakoids. To further advance the application of these polymers for studying biomembrane organization, we have examined the composition of thylakoid supramolecular protein complexes extracted by the five SMA polymers mentioned above. Two commonly studied plants, spinach (Spinacia oleracea) and pea (Pisum sativum), were used for extraction as model biomembranes. We found that the pSMAs differentially extract protein complexes from spinach and pea thylakoids. Based on their differential activity, which correlates with the polymer chemical structure, pSMAs can be divided into two groups: unfunctionalized polymers and ester derivatives.

Optimizing homogenizer-assisted extraction of chlorophylls from plantain epicarp (Musa paradisiaca L.)

The objective of this study was to establish parameters of optimization of the factors such as solid/solvent ratio, stirring speed and extraction time in homogenizer-assisted extraction (HAE) of chlorophyll a, b and total from plantain epicarp (Musa paradisiaca L.) using the Box–Benhken design. The individual effect and the interactions of the process variables (solid/solvent ratio, stirring speed (revolutions per minute, rpm), and extraction time in HAE) on the chlorophyll a, b and total contents in the extract were studied. The optimization was carried out with a Derringer desirability function, and it was found that the optimal extraction conditions included a solid/solvent ratio of 0.025 g/mL, an stirring speed of 13,508 and 90 s of treatment with Ultra-turrax. The corresponding predicted values were 33.67 mg/100 g of chlorophyll a, 32.915 mg/100 g of chlorophyll b and 66.585 mg/100 g of total chlorophyll. Finally, the optimal conditions were experimentally verified, and it was established that extraction with an Ultra-Turrax treatment is more efficient than the conventional method (maceration).

Air Pollution Tolerance Index of Some Tree Species of Pashupati and Budhanilkantha Area, Kathmandu

The present study aims to assess the air pollution tolerant trees from the roadside of Pashupati area of Kathmandu exposed to vehicular air pollution. This area with heavy traffic density was considered as the polluted site and Budhanilkantha, lying at the outskirt of Kathmandu with very less traffic density was considered as the less-polluted site and was selected as the control site of the study. Commonly available 21 roadside same tree species from both polluted and control sites were chosen. Air pollution tolerance index (APTI) values of the trees were calculated considering the biochemical parameters - relative water content, total leaf chlorophyll, ascorbic acid and leaf extract pH by using standard method. Significantly higher APTI values (P ≤ 0.05) were obtained in Cinnamomum camphora, Ficus elastica, F. religiosa, F. benghalensis and Grevillea robusta indicating that they are tolerant tree species. Phyllanthus emblica and Schima wallichii were found to be sensitive tree species.

Extraction of chlorophyll from wild and farmed Ulva spp. using aqueous solutions of ionic liquids

Products extracted from natural resources are an increasing trend in several fields promoted by consumer demand. Allied to the importance attached to the concept of “natural product” should be the way the “natural product” is obtained. In this work, chlorophyll was extracted from batches of wild-harvested and farm-raised green macroalgae Ulva spp. from two different European locations, Portugal and France. The performance of different aqueous solutions of tensioactive compounds such as ionic liquids and common surfactants in the yield of extraction of chlorophyll was studied and the operational conditions of extraction were optimized. The effect of drying the biomass on the yield of extraction of chlorophyll was evaluated as well as the effect of both locations (and the specific conditions of each location in terms of nutrients, water temperature and light intensity) in chlorophyll production. After optimization of all operational conditions, a maximum yield of extraction of 5.96 mgchl·gdry algae−1 was obtained using 250 mM of tributyltetradecylphosphonium chloride ([P4,4,4,14]Cl). The use of this solvent has allowed the development of a cost-effective (conclusion obtained after the economic analysis) and efficient process capable of maintaining the stability of the final product for more than one month.

Temperature Increase Research of Particles Formed by Ultrasonic Grinding of Spinach Leaves

The researchers chose spinach leaves as the research object for obtaining a natural dye based on chlorophyll because the vegetable is a food raw material rich in this pigment, and biologically active substances source with variety of its functional properties. To solve the problem of more complete chlorophyll extraction from crushed spinach leaves, they are to subject to a preliminary short-term soaking in a water/organic reagent emulsion under ultrasonic influence. Given that the water-based spinach suspension is thermolabile, a man must observe temperature restrictions, since when a certain critical temperature value is exceeded, the chlorophyll decomposition probability with its consumer properties loss is high. As it is practically impossible to determine the temperature inside a fine particle suspended in an aqueous emulsion experimentally, the authors considered to model internal and external heat transfer by solving the differential equation of heat energy transfer under appropriate boundary conditions using the numerical method of finite differences. The research determined structural-mechanical and thermophysical characteristics of crushed spinach leaves experimentally using the complex calculation method of thermophysical parameters based on the thermal inertia properties of a thermocouple. A man verified the solution adequacy by empirically determined suspension temperature after completion of the technological procedure. According to the developed algorithm, the authors compiled a program for the temperature fields evolution in the object of study considering the incident ultrasonic flow and an internal heat source with the new surface formation when grinding spinach particles in order to comply with thermal restrictions and maximize the quality parameters preservation. The article considers the rational duration of the ultrasonic grinding process without exceeding the recommended threshold temperatures.

Extraction of chlorophylls and carotenoids from dry and wet biomass of isolated Chlorella Thermophila: Optimization of process parameters and modelling by artificial neural network

Chlorophylls and carotenoids can be extracted from microalgae using various solvents. However, there is lack of studies regarding the comparison of extraction yield of these pigments from wet and dry microalgal biomass using different combination of cell disruption methods. Therefore, in this work, we have investigated the comparison of the extraction yield of chlorophylls and carotenoids from the wet and heat-dried microalgal biomass (isolated Chlorella thermophila) using ethanol. Extraction parameters such as homogenisation time, homogenisation speed, solvent temperature, solid-solvent ratio, boiling time and microwave time have been optimised. Chlorophyll extraction yield was observed to be 2.7 fold higher from wet biomass than dry biomass while carotenoid yield was 6.7 fold higher. Highest chlorophyll yield (∼60 mg/g-dry biomass) was observed at 6 min of homogenisation time, 10,000 rpm, solid solvent ratio of 1 mg/mL and 58 °C of solvent temperature from wet biomass with extraction efficiency of ∼94 %. Highest carotenoid yield was noticed following the same conditions of chlorophyll extraction except 4 °C of solvent temperature. The modelling of the extraction process was performed using artificial neural network (ANN) which may be useful for the scale-up of the extraction process at the industrial level.

Pulsed electric energy and ultrasonication assisted green solvent extraction of bio-molecules from different microalgal species

The effects of physical treatments (pulsed electrical fields (PEF), high voltage electrical discharges (HVED) and ultrasonication (US)) on aqueous extraction of carbohydrates and proteins, and ethanolic extraction of chlorophyll a from three microalgal species (Nannochloropsis sp., P. tricornutum and P. kessleri) have been studied. The total energy consumption of 530 kJ/kg suspension was applied for each treatment. For studied species, HVED was the most effective for extraction of carbohydrates, while US was the most effective for extraction of proteins and chlorophyll a. The observed differences for studied species can reflect the more fragile cell wall structure for P. tricornutum as compared with Nannochloropsis sp. or P. kessleri. The applied PEF, HVED and US treatments along with combinations of aqueous extraction of carbohydrates and proteins, and ethanolic extraction of pigments can be used in future implementations of selective extraction of valuable bio-molecules from microalgae.

Methane production through anaerobic digestion of residual microalgal biomass after the extraction of valuable compounds

Biorefinery concepts that combine several valorization pathways, including both bioproducts and bioenergy/biofuels, are of high interest in order to maximize the economic value of microalgal cultivation systems. In this study, the residual biomass obtained from the microalga Chlorella vulgaris following the chemical extraction of a valuable fraction consisting of either chlorophyll, proteins, or lipids was used as feedstock for biogas production in a laboratory-scale biochemical methane potential (BMP) assay performed in batch mode. The main aim of this study is to assess the effect of chemical extraction and removal of valuable fractions on the methane yield of residual microalgal biomass. The dry weight ratio of residual biomass to total biomass was 0.54 g/g after lipid extraction, 0.62 g/g after chlorophyll extraction, and 0.42 g/g after protein extraction. The methane yields of raw microalgae as well as residual microalgal biomass after lipid, chlorophyll, or protein extraction and removal were in the range 207–237 mL CH4/g volatile solids. Differences in the methane yields were not statistically significant. Hence, the recovery and removal of valuable compounds did not have a significant impact on the final methane yields of microalgal biomass by anaerobic digestion. Nevertheless, the methane production kinetics of residual microalgal biomass after lipid extraction displayed a diauxic pattern, possibly due to the inhibition of methanogenic bacteria by chloroform used as extraction solvent. This suggests that solvent toxicity must be considered when applying anaerobic digestion as a downstream process in biomass-based biorefineries.

Estimation and comparison of effective compounds in two algae species identified in Qeshm Island (Persian Gulf)

Estimation and comparison of effective compounds (mineral contents, amino acid compositions, fatty acid distributions, protein, fat and total fiber contents) in two algal species Sirophysalis trinodis and Polycladia myrica collected from Qeshm Island were done. They after washing with tap water were dried in an oven at 40 ◦C for 3 days. The dried algae samples were pulverized and packed in airtight plastic bags. The results showed that the marine macro-algae included low lipid contents about 0.03±0.01 and 0.33±0.10% dry sample weight and proved to be a rich source of fatty acid (Palmitic and Oleic acid) and minerals. The protein content of these brown seaweeds was lower than 15%. All essential amino acids were detected in the seaweed species, featured uniquely high concentrations of amino acid when compared to ground plants. Also, results showed that the chlorophyll amount dependent on solvent type and cell wall structure of algae. Hence, in this study, among the three acetone solvents, diethyl ether and methanol, acetone showed the best performance for the extraction of chlorophyll in both algae S. trinodis and P. myrica. In general, the aim of this study was to determine the valuable compounds of two abundant species of brown algae in the south of the country and introduce them to food units.

Molecular functional group and optical analysis on chlorophyll of green choy sum and cassava leaves extracts

Recently, chlorophyll can improve the optical characteristics of thin films. The role of chlorophyll is a dopant in the thin film. Chlorophyll is very good in transferring electron which is strongly related to energy. If the energy gap of the film is low, the characteristics of the film are better because it requires less energy to jump one electron from the valence band to the conduction band. However, an important factor to consider in this case is the chlorophyll content. The more chlorophyll content will improve the light absorption rate. This study examined the characterization of chlorophyll of green choy sum and cassava leaves extracts. The analysis was carried out based on the Fourier Transform Infra-Red (FTIR) and UV-Vis Spectrophotometer. The results of the analysis were molecular functional groups, vibration frequencies, bond force constants between molecules, optical characteristics and chlorophyll content on both of chlorophyll extracts.

Solvent Extraction Optimization of Chlorophyll Dye from Conocarpus Lancifolius Leaves

Recently, the green food industry has started using chlorophyll dye as a healthy and safe color additive for food and beverages. In this study, a conventional extraction technique was used to extract chlorophyll dye from Conocarpus lancifolius leaves, using 80% acetone as solvent. Taguchi's L25 orthogonal array was utilized to optimize the extraction parameters with total chlorophyll dye yield as the response. The optimum extraction conditions were 75 minutes of extraction time, 33.5 °C extraction temperature and a 118 ml/g solvent to raw leaves ratio. Analysis of variance using Minitab18 was conducted to identify the most significant extraction parameters and available interactions. It was found that the chlorophyll dye yield from Conocarpus lancifolius leaves is mostly dependent on extraction time, followed by mixing ratio and temperature. The analysis showed a large interaction between the extraction parameters studied. The experimental chlorophyll dye yield was in close agreement with the predicted model. The adequacy of the predicted model was confirmed by an experiment conducted under the optimal conditions, given a 3.9% error percentage.

Integrative process to extract chlorophyll and purify rosmarinic acid from rosemary leaves (Rosmarinus officialis)

AbstractBACKGROUNDRosemary leaves have been widely employed in the food and pharmaceutical industry due to the presence of important compounds. However, during the extraction process, in addition to the target biomolecules [chlorophyll (Chl), and rosmarinic acid], other components (phenolic compounds) are also extracted. This work focuses on the development of a sequential and selective protocol for the extraction and concentration of Chl using an aqueous micellar two‐phase system, and extraction and purification of rosmarinic acid using an aqueous two‐phase system (ATPS).RESULTSFirstly, the extraction and purification of Chls were studied. The Chls rich‐extract obtained using the best conditions of aqueous solution of Triton X‐100 (5 wt%) and phosphate buffer (50 mmol L−1), 50 °C solid–liquid ratio was evaluated at 1:100 and 60 min (Chl a = 69.3 ± 1.9 mg 100 g−1; Chl b = 41.1 ± 1.2 mg 100 g−1 and total Chl = 110.5 ± 3.1 mg 100 g−1) were submitted to temperature above cloud point. The Chl b and total Chl were concentrated 3.1 and 1.7 times. Further, the extraction and purification of rosmarinic acid were investigated. Ethanol, methanol, and acetonitrile (80 wt%) at 50°C were investigated; and after extraction, the rosmarinic acid rich‐extract were add in the ATPS composed of organic solvent + tripotassium phosphate (K3PO4). The extraction efficiencies were observed in the top phase (100%) for all systems. The highest purification factor was found using the ATPS based on acetonitrile (50 wt%) + K3PO4 (20 wt%).CONCLUSIONSThe sequential protocols to extract Chls and purify rosmarinic acid can be applied. © 2020 Society of Chemical Industry

Two-step procedure for selective recovery of bio-molecules from microalga Nannochloropsis oculata assisted by high voltage electrical discharges

Two-step procedure with the initial aqueous extraction from raw microalgae Nannochloropsis oculata and secondary organic solvent extraction from vacuum dried (VD) microalgae were applied for selective recovery of bio-molecules. The effects of preliminary aqueous washing and high voltage electrical discharges (HVED, 40 kV/cm, 4 ms pulses) were tested. The positive effects of HVED treatment and washing on selectivity of aqueous extraction of ionics and other water-soluble compounds (carbohydrates, proteins and pigments) were observed. Moreover, the HVED treatment allowed improving the kinetic of vacuum drying, and significant effects of HVED treatment on organic solvent extraction of chlorophylls, carotenoids and lipids were determined. The proposed two-step procedure combining the preliminary washing, HVED treatment and aqueous/organic solvents extraction steps are useful for selective extraction of different bio-molecules from microalgae biomass.

Bio-refinery of Chlorella sorokiniana with pulsed electric field pre-treatment

The aim of this work was to investigate the potential of PEF technology for green extraction of microalgal pigments and lipids from fresh Chlorella sorokiniana suspensions. Efficiencies of PEF treatment and different solvent systems application to C.sorokiniana were compared to efficiencies of untreated biomass extraction. Differences in chlorophyll extraction of untreated and PEF treated C.sorokiniana were only seen at short extraction times. Beneficial PEF-effect was minimised for long-time extractions of larger algae quantities where yields aligned. Extraction attempts on C. sorokiniana lipids did not show increased extractability after PEF treatment, which underlined the statement of PEF representing a rather ineffective disruption method for microalgae holding rigid cell walls.

Study of Behavioral Changes and Photosynthetic Activity of &lt;i&gt;Euglenas gracilis&lt;/i&gt; in the Presence of Effluents from the Laboratory of Clinical Analysis

Healthcare waste has now been increasingly studied in terms of the risks or dangers that can cause the environment and human health. Waste generated in clinical analysis laboratories (CALs) deserves attention, because, due to the advent of the concept of emergent pollution, it is doubtful if the materials or reagents are disposed in the sewage by CALs, which are currently considered non-contaminated or with low risk potential, under current legislation, may actually impact the environment with actions not yet understood. This study was experimental and conducted at the Environmental Laboratory of the University of the Region of Joinville. It was used Euglena gracilis (primary trophic level) algae exposed to effluents from five sectors of a CAL: Biochemistry, Hematology, Viral Load, Tuberculosis and Immunochemistry. Samples were collected from the siphons attached to the wash sinks of the CAL materials. To verify changes in algae that denote environmental danger, behavioral changes were analyzed via NGTOX, and chlorophyll concentration was calculated by chlorophyll extraction according to Mendel’s method. Viral Load (VL) and Hematology (HT) sectors were the ones that most affected algae (Tukey test). In both sectors, there was inhibition of algae mobility and gravitaxy: in VL, due to the presence of chaotropic agents that denature organic structures; and in HT, due to the change in membrane permeability attributed to methylene blue. Also in HT, there was a search for algae adaptation by increasing the rise to the surface in order to overcome the lower luminosity due to the coloration of the environment, which also affects photosynthesis. Regarding the concentration of a-chlorophyll, the VL and HT were the most affected as well, being the first one the one that had more concentration reduction because of the presence of chaotropic agents. Considering new parameters evaluated, the discarded compounds need to be better evaluated for risk, as they affect algal photosynthesis. Procedures for removal of these compounds should be considered.

Cellulose fibers extraction from Ulva lactuca from the Black Sea

Abstract Cellulose fibres are known for their good mechanical properties, therefore they are used as fillers in structural composite materials, including as nanofibrils in nanomaterials. Also, they are biocompatible, non-toxic and biodegradable, reason for their use in the food industry as packaging materials or in obtaining medical materials. One source of cheap, easy- to- extract cellulose is the algal mass of Ulva lactuca, one of the most frequent species found in the Black Sea. In this study, cellulose extraction from Ulva lactuca was achieved by a simple low cost physical-chemical treatment. Freshly harvested seaweed was dried at 45 °C for 48 hours, transformed into a fine powder in order to increase the contact surface between the solvents and the alga. Extraction of lipids and chlorophyll took place in Soxhlet apparatus with ethanol. Successive steps of chemical treatment, having in view removal of hydrosoluble ulvans, pigments and hemicellulose lead to a yield of 15.36% in dry matter (DM) of cellulose-rich insoluble fraction proving that Ulva Lactuca species is a viable alternative resource in cellulose production.

Экстракция растительных пигментов из местного сырья

Introduction. Violation of the ecological balance and modern processing methods may result in a lower content of important nutrients in food. Therefore, there is a need to restore the balance of biologically active substances in food products. In this regard, it seems appropriate to use plant materials that contain biologically active substances, as well as pigments. The use of aromatic plants for these purposes remains understudied. The research objective was to extract chlorophyll-based green pigments based from green plants.&#x0D; Study objects and methods. The object of the study was dried parsley (Petroselinum vulgare). The extractants included acetone, ethyl alcohol (95%), hexane, petroleum ether, and chloroform.&#x0D; Results and discussion. The extractants were chosen according to the total content of chlorophylls, which determine the color of the extract. Since carotenoids are extracted along with chlorophylls, the content of this component was also determined. Acetone demonstrated the maximum degree of chlorophyll recovery. However, this solvent cannot be used in food industry. Therefore, 95% ethanol, which also showed a high degree of chlorophyll extraction, was chosen as the extractant. A set of experiments made it possible to establish the ratio of feedstock and extractant as 1:60 (three times extraction with a mass ratio of parsley and extract); the optimal time of each extraction was 0.5 ± 0.1 h. Magnesium carbonate (MgCO2) helped to preserve the dyes. In order to increase the concentration of coloring substances, the obtained extract was subjected to thickening at a residual pressure of 400 Pa at 40°C.&#x0D; Conclusion. The obtained concentrate from dried parsley can be used in production of combined foods.

Natural dyeing of merino wool fibers with Cinnamomum camphora leaves extract with mordants of biological origin: a greener approach of textile coloration

In this research work, an attempt has been made to investigate the possibility of wool dyeing with natural dye extracted from the Cinnamomum camphora middle aged/mature leaves. Visually, a range of ecofriendly shades with quite different color gamut of varying hue and tone were produced via the application of different natural mordants. The effect of various natural mordants (gallnut, pomegranate peel, arjun bark, chlorophyll extract, and citric acid) and some commonly used metal mordants (ferrous sulfate, copper sulfate, stannous chloride, and sodium dichromate) on color and fastness properties of dyed wool samples was comparatively evaluated in conjunction with 50% (o.w.f.) of C. camphora natural dye. Dyeing experiments were performed with and without mordants employing pre-mordanting technique. The color of dyed wool fibers were investigated in terms of CIELab (L*, a*, and b*) and color strength values (K/S); and fastness properties were determined as per ISO standard test methods. In general, pre-biomordanted wool fibers with P. granatum (3, 4 and 5% o.w.f.), citric acid (5% o.w.f.) and chlorophyll extract (10–50% o.w.f.) showed comparable color and fastness results compared to those of metal treated samples. Durability of color was also checked to assess the effect of washing of dyed wool fibers. This research work on the use of ecofriendly biomordants will provide a valuable reference in future for ecological and environmentally safe dyeing.