Chlorophyll Extraction Research Articles

Preparation of Chlorophyll Nanoemulsion from Pomelo Leaves and Its Inhibition Effect on Melanoma Cells A375.

Pomelo (Citrus grandis), an important fruit crop grown in tropical and subtropical areas, is cultivated mainly in Asian countries. The dominant pigment in pomelo leaves, chlorophyll, has been reported to possess many biological activities such as antioxidant, anti-inflammation and anticancer. The objectives of this study were to determine chlorophylls in Pomelo leaves by high-performance liquid chromatography-mass spectrometry (HPLC-MS) and to encapsulate the isolated chlorophylls from preparative column chromatography into a nanoemulsion system for elucidating the inhibition mechanism on the growth of melanoma cells A375. The results showed that chlorophyll a and chlorophyll b could be separated within 25 min by using a C18 column and a gradient ternary mobile phase of acetone, acetonitrile and methanol. Pomelo leaves mainly contained chlorophyll a (2278.3 μg/g) and chlorophyll b (785.8 μg/g). A highly stable chlorophyll nanoemulsion was prepared with the mean particle size being 13.2 nm as determined by a dynamic light scattering (DLS) method. The encapsulation efficiency of chlorophyll nanoemulsion was 99%, while the zeta potential was −64.4 mV. In addition, the chlorophyll nanoemulsion possessed high thermal stability up to 100 °C and remained stable over a 90-day storage period at 4 °C. Western blot analysis revealed that chlorophyll nanoemulsion and extract could upregulate p53, p21, cyclin B and cyclin A as well as downregulate CDK1 and CDK2 in a concentration-dependent manner for inhibition of melanoma cells A375. Furthermore, chlorophyll nanoemulsion and extract could upregulate Bax and cytochrome C and downregulate Bcl-2, leading to activation of caspase-9, caspase-8 and caspase-3 for the induction of cell apoptosis. Compared to chlorophyll extract, chlorophyll nanoemulsion was more effective in inhibiting the growth of melanoma cells A375.

Supercritical fluid (CO2+ethanol) extraction of chlorophylls and carotenoids from Chlorella sorokiniana: COSMO-SAC assisted prediction of properties and experimental approach

In this work, COSMO-SAC was used to calculate infinite dilution activity coefficients (γ∞) of carotenoids in CO2+ethanol mixtures as a theoretical approach to evaluate the effect of solvent composition on solute solubility. In an experimental approach, a central composite design was employed to evaluate the use of supercritical mixtures of CO2+ethanol to extract chlorophylls and carotenoids from the microalga Chlorella sorokiniana. The carotenoid profiles of extracts were fully evaluated, and the optimal conditions for the extraction of chlorophyll a, chlorophyll b, carotenes, lutein, zeaxanthin and violaxanthin were differentiated. Extraction temperature, pressure and the composition of the solvent proved to be relevant factors, as the experimental conditions rendered fundamentally different extraction results. The presence of ethanol proved to be essential for the efficient extraction of chlorophylls and to substantially improve the recovery of carotenoids. Scanning electron microscopy (SEM) was used to evaluate cell integrity, and indicated that the increase of pressure may have positively affected the extraction of compounds by causing disruption of the particles. The theoretical approach in this work was able to corroborate experimental results for the extraction of carotenoids from C. sorokiniana, reinforcing the hypothesis that carotenoids are extracted from the microalgal biomass as aggregates, and showing that COSMO-SAC is an important tool for the selection of the supercritical solvent.

Green solvent-based extraction of chlorophyll a from Nannochloropsis sp. Using 2,3-butanediol

Chlorophyll acts as a beneficial antioxidant for the human body; however, only a few studies have reported the chlorophyll extraction using the non-toxic solvents. In this study, we used a green solvent, 2,3-butanediol (2,3-BDO), for extraction of chlorophyll a from Nannochloropsis sp. for the first time. Levo-isomer of 2,3-BDO exhibited remarkable chlorophyll extraction properties compared to its meso-isomer, and other conventional solvents such as ethanol. Hydrogen bonding between the solvent and chlorophyll was a detrimental factor for evaluating the extraction efficiency. The extraction yield reached 91.8% with the maximum specific antioxidant activity at 10,000 rpm for 20 min, using 25 mL solvent/g dry cell with assistance of a high shear mixer (HSM). The scalability was verified upon comparing the lab- and bench-scale tests, which demonstrated similar extraction yields and antioxidant activities, reducing process time and specific energy consumption.

Microwave-assisted extraction of chlorophyll and polyphenol with antioxidant activity from Pandanus amaryllifolius Roxb. in Vietnam

Pandanus amaryllifolius Roxb., also called as pandan leaf, is distributed mainly in hot and humid climate areas. Pandan leaf, grown very popular in Vietnam, contains many biological value compounds such as chlorophyll and polyphenol with oxidation prevention activity. Microwave-assisted extraction (MAE) has advantages such as reduced extraction time and high extraction efficiency in comprison to other extraction technologies. The aim of this study was to assess the effect of microwave-assisted extraction conditions on chlorophyll and polyphenol content as well as antioxidant activity of pandan leaf. Research results show that acetone solvents of 90°, pandan powder: acetone ratio of 1:30, microwave capacity of 300W, microwave-assisted time of 2 minutes are parameters for the highest extraction efficiency. With these conditions, chlorophyll a, b and total chlorophyll content are respectively 9.4278 μg/mL, 4.2460 μg/mL, 13.6738 μg/mL, polyphenol content is 2.7577 g/L and DPPH radical collection activity 1409.51 μmol/L.

A comparative study of the growth of microalgae-bacteria symbiotic consortium with the axenic culture of microalgae in dairy wastewater through extraction and quantification of chlorophyll

Microalgae-bacteria symbiotic association can provide many competitive advantages over the pure axenic culture of microalgae to obtain biofuel. This system significantly reduces an important technical constraint, including associated capital cost with the infrastructure required for maintaining pure microalgae culture. The current study aims to compare the symbiotic growth of Tetraselmis indica - Pseudomonas aeruginosa consortium with axenic growth of Tetraselmis indica in dairy wastewater (DWW). The extraction and optimization of chlorophyll-a were also investigated. The overall maximum chlorophyll-a recovery of 29.106 ± 0.045 mg L−1 was obtained through the sonification as extractant method with methanol as an extraction solvent. The chlorophyll-a content of consortium cultivated in DWW higher than the axenic pure microalgae culture (APMC). The maximum biomass yield of the consortium was achieved as 1454.88 mg L−1, which is much higher than APMC growth in DWW by 38.80%. On the 10th day, the consortium removed 87.49%, 83.76%, and 79.83% of chemical oxygen demand (COD), total dissolved nitrogen (TDN), and total dissolved phosphorus respectively. The higher biomass growth and wastewater nutrient removal exhibited by the symbiotic microalgae-bacteria consortium can be utilized in biofuel technology.

Kinetic Study on Chlorophyll and Antioxidant Activity from Polyscias fruticosa (L.) Harms Leaves via Microwave-Assisted Extraction.

Polyscias fruticosa (L.) leaves contain significant bioactive compounds with high antioxidant activity such as chlorophylls, total polyphenols, etc. but these have still been underutilized. In this study, the kinetics of chlorophyll and antioxidant activity extraction from P. fruticosa leaves by microwave-assisted extraction (MAE) were investigated. Microwave power was 300, 450, or 600 (W); the ratio of material/solvent varied from 1:40 to 1:80 (g/mL). In this study, the second-order kinetic model successfully predicted the change of chlorophyll and antioxidant activity during MAE. The increase of microwave power or/and the solvent amount increased saturated extraction efficiency and the extraction rate constant. However, the saturated concentration of chlorophyll and antioxidant activity increased with the increment of microwave power and the decrease in solvent amount.

Estimating Leaf Chlorophyll Content of Buffaloberry Using Normalized Difference Vegetation Index Sensors

Commercial optical chlorophyll meters estimate relative chlorophyll content using the ratio of transmitted red light and near-infrared (NIR) light emitted from a red light-emitting diode (LED) and an NIR LED. Normalized difference vegetation index (NDVI) sensors have red and NIR light detectors and may be used to estimate chlorophyll content by detecting the transmitted red and NIR light through leaves. In this study, leaf chlorophyll content of ‘Torrey’ buffaloberry (Shepherdia ×utahensis) plants treated with 0 mm [zero nitrogen (N)], 2 mm (medium N), or 4 mm (ample N) ammonium nitrate for 3 weeks were evaluated using two commercial chlorophyll meters and NDVI sensors. The absolute chlorophyll content was determined using chlorophyll extraction. Our results showed that plants receiving ample N and medium N had decreased transmitted red light (i.e., greater absorption in red light). Measurements of optical chlorophyll meters, NDVI sensors, and chlorophyll extraction similarly showed that plants receiving medium N and ample N had greater leaf chlorophyll content than those receiving zero N. Relative leaf chlorophyll content estimated using NDVI sensors correlated positively with those from the chlorophyll meters (P < 0.0001; r2 range, 0.56–0.82). Therefore, our results indicate that NDVI measurements are sensitive to leaf chlorophyll content. These NDVI sensors, or specialized sensors developed using similar principles, can be used to estimate the relative chlorophyll content of nursery crops and help growers adjust fertilization to improve plant growth and nutrient status.

Photocatalytic reduction of Cr(VI) within mesoporous TiO2 templated and confined with chlorophyll

AbstractBiomimetic light‐harvesting architecture has been proposed as an alternative for existing solar conversion systems. Chlorophyll‐sensitized TiO2 have been identified as promising catalyst for photocatalytic reduction system, but it is largely limited to complicated process for purification of chlorophyll and adding extra hole scavenger in system. In sharp contrast to conventional photosensitization methods in which the organic dyes were often adsorbed after photosensitization, herein mesoporous TiO2 incorporated with chlorophyll extract was synthesized in situ (TiO2/Chl). Interestingly, TiO2 incorporated with chlorophyll exhibited two distinct absorption bands around 407 and 668 nm which are closed to the characteristic absorption peak of chlorophyll. TiO2 incorporated with chlorophyll kept at 185°C for 1.5 hours (TiO2/Chl‐185) achieved a high photoreduction rate of 86.1% for the photocatalytic reduction of Cr(VI), which was approximately 29 and 20 times higher than that of TiO2 and TiO2 adsorbing chlorophyll (TiO2/ads Chl), respectively. The significant enhancement was due to the confinement of chlorophyll. This demonstrates that mesoporous TiO2 templated and confined with chlorophyll could offer a new approach for synergistic photoreduction of Cr(VI).

Optimization of Glycerol-Water Extraction of Selected Bioactive Compounds from Peppermint and Common Nettle.

Current trends in the industry indicate that extraction solvents should conform with the ideals of so-called “green chemistry”. Therefore, the objective of the presented study was to optimize the conditions for the extraction of polyphenols, flavonoids and chlorophyll from peppermint leaves (Mentha × piperita L., Lamiaceae) and from common nettle leaves (Urtica dioica L., Urticaceae) via green chemistry. The obtained experimental results were subjected to modelling by means of the multiple regression method, while the optimization of the system was addressed via the application of the desirability function. As a result of the use of glycerol–water systems for the extraction of the tested active compounds from mint leaves and nettle leaves, extracts with higher concentrations of polyphenols, flavonoids and chlorophyll were most often obtained, when compared with the use of classical solvents such as water and ethanol. In this work, we demonstrate that the extraction temperature has significant influence on the concentration of the determined components in the extracts. To obtain the highest values of the analysed parameters, leaves of peppermint should be extracted with glycerol–water mixture at the proportions of 30.5:69.5 at a temperature of 50 °C, while the optimal conditions for the extraction of leaves of common nettle were the glycerol–water proportions of 12.5:87.5 and extraction temperature of 20 °C. Comparing the average percentage differences between the highest values of the analysed parameters obtained in the experiment and the approximated values for various temperatures with the level of desirability, one can note a high correlation that, in the analysed examples, amounted to 0.9681. The study showed that glycerol can be an alternative solvent in the extraction of polyphenols, flavonoids and chlorophyll, replacing, e.g., ethanol—which, for various reasons, cannot always be used.

Extraction of Chlorophylls and Carotenoids from Microalgae: COSMO‐SAC‐Assisted Solvent Screening

AbstractEfficient pigment extraction is a key factor influencing the economic feasibility of microalgal biorefineries. Ethanol, acetone, ethyl acetate, and hexane were evaluated for the batch extraction of chlorophyll a, chlorophyll b, and carotenoids from Chlorella sorokiniana and Heterochlorella luteoviridis. Solvent‐to‐biomass ratios did not affect extraction results in most cases. Ethanol showed the highest efficiency in the recovery of chlorophylls, presenting high extraction yields. Acetone was among the most efficient solvents to extract violaxanthin, lutein, zeaxanthin, and carotene from both microalgae, proving to be generally suitable to this application. Conductor‐like screening model for real solvent segment activity (COSMO‐SAC) corroborated experimental results, proving to be an important tool in the screening of solvents for extraction processes.

Extraction and Fractionation of Pigments from Saccharina latissima (Linnaeus, 2006) Using an Ionic Liquid + Oil + Water System

There is a strong industrial interest in the development of greener and more sustainable processes based on the use of renewable resources, and a biorefinery based on marine resources, such as macroalgae, stands as a major opportunity toward that end. In this work, Saccharina latissima (Linnaeus), a brown macroalga, was used as a source of pigments to develop an integrated platform that is able to promote the extraction and separation of chlorophyll and fucoxanthin in one single step. The process was studied, and its operational conditions were optimized with yields of extraction of chlorophyll and fucoxanthin of 4.93 ± 0.22 mgcₕₗ·gdᵣy bᵢₒₘₐₛₛ–¹ and 1956 ± 84 μgfᵤcₒ·gdᵣy bᵢₒₘₐₛₛ–¹, respectively. These results were achieved with extraction systems composed of 84% of an aqueous solution of a tensioactive phosphonium-based ionic liquid (IL) at 350 mM + 16% of sunflower oil, during 40 min, using a solid–liquid ratio of 0.017 gdᵣy bᵢₒₘₐₛₛ·mLₛₒₗᵥₑₙₜ–¹. After the separation of both aqueous IL-rich and oil-rich phases, the IL content in both phases was investigated, the oil phase being free of IL. Envisioning the industrial potential of the process developed in this work, the recovery of the IL from the aqueous IL-rich phase of the initial system was attempted by a back-extraction using organic solvents immiscible in water, being shown that 82% of the IL can be recovered and reused in new cycles of extraction. The environmental and economic impacts of the final process proposed for the extraction and fractionation of chlorophyll and fucoxanthin were evaluated. Different scenarios were considered, but summing up the main results, the solvents’ recycling allowed better results, proving the economic and environmental viability of the overall process.

Continuous extraction of Spirulina platensis biopigments using different extraction sequences

The use of microalga Spirulina is developing rapidly in the industry. Spirulina biomass is widely used in the food, pharmaceutical and cosmeceutical industry because it is rich in biochemicals and antioxidants that are beneficial to human health. This study aimed to determine the biopigment contents in S. platensis and examine the sequences of continuous extraction process to obtain the optimal biopigment yields. Biopigments of S. platensis extracted in this study were total chlorophyll of 52.28 μg/mL (25.28 μg/mL chlorophyll a and 27 μg/mL chlorophyll b), total carotenoid of 7.9 μg/mL and phycocyanin of 0.020 mg/L. In addition to biopigments, a residue in the form of solid powder was also produced at 20.7 % of biomass. Simultaneous extraction of chlorophyll and total carotenoids was successfully applied in this study, while the optimal phycocyanin extraction should be further developed. The optimal extraction sequence of biopigments was extraction of chlorophyll then carotenoid with low heating and followed by extraction of phycocyanin.

Development of a new method for simultaneous extraction of chlorophylls and carotenoids from microalgal biomass

There are still limitations in the pigment extraction methods used in microalgae biomass, especially for laboratory scale. This work aimed to develop a simple method for simultaneous extraction of chlorophylls and carotenoids from the microalgae Spirulina sp. and Scenedesmus obliquus. The extraction was performed using a methanol/chloroform (2:1 v/v) mixture with different homogenization times (30, 60 and 120 min); after exhaustive extraction with chloroform, acetone, ethyl acetate, and ethanol were tested and compared with the reference maceration method (MM). Satisfactory recovery values, mostly between 101.1 and 124.2%, were obtained with 30 min of homogenization and extraction with ethanol after chloroform (PM30E). In general, the chlorophyll and carotenoid profiles obtained by the PM30E presented fewer isomers and possible degradation compounds. PM30E showed a reduction of about five times the volume of organic solvents and was four times faster than the MM. In addition, this method provided greater analytical frequency and security for the analyst, since PM30E is performed in a closed system of mechanical homogenization.

Extraction of chlorophyll and carotenoids loaded into chitosan as potential targeted therapy and bio imaging agents for breast carcinoma

In the current study, the treatment efficacy of ECHCAH was evaluated in vitro studies using cell viability and flow cytometry in human TNBCs. The results here showed significant gradual reduction in growth of TNBCs (MDA-231cell lines) after their exposure to serial concentrations for hydrogel assembly (5 μg/mL to 25 μg/mL) for 24 and 48 h, representing (86 ± 1% to 45 ± 1.5% p < 0.001) and (79 ± 1.5% to 35 ± 2.5% p < 0.001) respectively. The flow cytometry showed significant increase in the present of late apoptotic and necrotic cells (64% ± 1.2 and 27% ± 0.3 p < 0.001) after 48 h incubation compared to untreated cells (1.13% ± 0.3 and 4% ± 0.2 p < 0.001) respectively. It can be summarized that ECHCA inside targeted hydrogel assemblies can inhibit proliferation of cancer cells.

Optimized extraction of neutral carbohydrates, crude lipids and photosynthetic pigments from the wet biomass of the microalga Scenedesmus obliquus BR003

The establishment of microalgae as a sustainable source of bioproducts requires a detailed evaluation of unit operations that compose a biorefinery. Biorefining of microalgal biomass is performed in sequence for obtaining a diversity of valuable compounds, but the optimum sequence of biorefining still under scrutiny. Herein, detailed modeling of unit operations was performed to enhance the extraction of neutral carbohydrates, crude lipids, and photosynthetic pigments from the wet biomass (slurry) of the microalga Scenedesmus obliquus BR003. Concentrated sulfuric acid hydrolysis resulted in an optimized extraction yield of 93% at temperature of 80 °C, 1.5 mol L−1 of sulfuric acid solution, and retention time of 120 s. Temperature and retention time were important parameters to increase the extraction of neutral carbohydrates using concentrated sulfuric acid. The efficiency of the multistage extraction of crude lipids was drastically decreased by high biomass-to-solvent ratios, and the use of proper mixing systems and longer mixing times increased the efficiency of this unit operation. The highest lipid content was achieved using orbital shaker (210.1 ± 10.9 mg g−1), retention time of 55 min, and biomass-to-solid ratio of 6.6% (w w−1). The extraction of chlorophylls a and b, and total carotenoids could be described by sigmoidal functions between the extraction yields and ratios of ethanol to the microalgal slurry, hence high levels of ethanol were required to efficiently extract them. The minimum ratio of ethanol to microalgae slurry necessary to efficiently extract photosynthetic pigments from the slurry of S. obliquus BR003 was 0.8:0.2 in volume, and this condition allowed the extraction resulted in extraction yields from 84 to 100%.

Utilization of leaf and fruit extracts of kedondong (Spondias dulcis Forst) as a supporting material for energy conversion in dye sensitized solar cells and electrochemical cells

This study aims to obtain a dye sensitized solar cell (DSSC) prototype and a voltaic cell prototype using the kedondong plant (Spondias dulcis Forst). Kedondong leaves as a source of chlorophyll were deliberately chosen to be in line with the use of kedondong fruit as a material for electrochemical cells, so that two research results could be obtained from the kedondong plant. This research is for the application of scientific development, increasing the added value of kedondong plant, and as support for the use of environmentally friendly energy. Research methods in general are chlorophyll extraction, citric acid isolation, chlorophyll testing, DSSC assembly, electrochemical cell assembly, and performance testing for both DSSC and electrochemical cells. The analysis results show that typical peaks appear for the functional groups in chlorophyll at certain wave numbers. Hydroxyl (-OH) groups appear in the range of 3650 - 3200 cm-1, the C=C group at 1700 - 1500 cm-1, the C-N group at 1350 - 1000 cm-1, and the C-O group at 1300 - 1000 cm-1. Quantitative testing used a UV-Vis spectrophotometer at 645 nm and 633 nm, the results obtained were chlorophyll a was 4.52 mg/L, chlorophyll b was 8.43 mg/L, and total chlorophyll was 12.95 mg/L. The DSSC prototype performance test with a size of only 2.5 cm2, can produce a minimum voltage of 30.1 mV and a maximum of 280 mV. The electrochemical test of kedondong citric acid extract showed that the value of the electric voltage ranged from 121 to 630 mV. Keywords: Kedondong, Chlorophyll, DSSC, Citric acid, Electrochemical cells

Chlorophyll Extraction Methods Review and Chlorophyll Stability of Katuk Leaves (Sauropus androgynous)

Chlorophyll is one of the most used coloring materials derived from green leaves and other parts of almost all green plants. It contains several components that are beneficial to health: vitamins, antioxidants, antibacterial, anti-inflammatory; hence, chlorophyll is a nutraceutical. Chlorophyll has been developed by extracting green plants, for instance, katuk leaves (Sauropus androgynous). A review of various chlorophyll extraction methods from natural ingredients was done to obtain the most suitable katuk leaves. Preliminary experiments had been carried out to develop the chlorophyll stability of katuk leaves on the influence of temperature, pH, and storage time. The chlorophyll content of katuk leaves decreased at high temperatures. Extraction using supercritical/subcritical CO2 can extract components selectively so that it is suitable for extraction applications for products that are not heat resistant. Meanwhile, carbon dioxide (CO2) as a supercritical/subcritical solvent has several advantages, including producing an inert gas that is non-toxic, non-flammable, and cheap. Under normal gas conditions, it is straightforward to separate CO2 and dissolved components without heating and producing extracts free of solvents. The most suitable chlorophyll extraction method of katuk leaves was supercritical/subcritical CO2 extraction because it could prevent chlorophyll degradation.

Recovery of Chlorophyll a Derivative from Spirulina maxima: Its Purification and Photosensitizing Potential

Spirulina sp. is a cyanobacterium rich in essential amino acids and pigments such as chlorophyll a, xanthophylls, and phycocyanin. Besides many other applications, chlorophyll a and its derivatives are being studied as photosensitizers in photodynamic therapy for cancer treatment. In this work, two methodologies of solid–liquid extraction were developed, and their performance compared; one using conventional organic solvents and the other using aqueous solutions of ionic liquids (ILs) and surfactants. It was found that an aqueous solution of an ammonium-based ionic liquid was able to increase the yield of extraction of chlorophyll a from Spirulina maxima in 25% compared with the conventional methodology using methanol. Besides, the proposed alternative methodology allows the separation of chlorophyll a from xanthophylls using a simple liquid–liquid extraction. The IL can be recovered by backextraction using ethyl acetate, while the chlorophyll derivative is shown to retain its ability to generate oxygen singlets, which is essential to its potential application as a photosensitizer in photodynamic therapy.

Suitability of four main Mediterranean tree crops for their growth in peri-urban agriculture and restoration (Gabes, Tunisia).

In order to increase the knowledge about crop tolerance to air pollutants in the different agroclimatic zones of the world, so that they can be efficiently considered for improving peri-urban agriculture, increasing the success of restoration projects, or enhancing air quality in polluted sites, the suitability of four economical valuable tree crops of the Mediterranean agriculture were studied under field conditions: date palm tree (Phoenix dactylifera L.), pomegranate (Punica granatum L.), fig tree (Ficus carica L.), and olive tree (Olea europaea L.). The measurement of biochemical markers such as ascorbic acid content, leaf relative water content, leaf total chlorophyll and leaf extract pH, at two contrasted air quality sites, a polluted site located around Gabes (Tunisia) industrial area and a control site, allowed the assessment of the air pollution tolerance index (APTI) and anticipated performance index (API) for the assayed species. Results showed obvious differences between the evergreen and the caducifolious tree crops assayed. Phoenix dactylifera tree (API = 6) was classified as an excellent performer for growing under poor air quality, followed by Olea europaea tree (API = 2) which was classified as a moderate performer. Both of trees can be recommended for successful results in peri-urban agriculture and restoration projects of polluted areas in the Mediterranean climate; on the contrary, the suitability of the Punica granatum (API = 1) was very poor, but still potentially interesting as a biological indicator of air pollution. Regarding the Ficus carica tree (API = 0), this species is not suitable for growing in air-polluted areas.

Development of an Ultrasonic-Assisted Extraction Technique for the Extraction of Natural Coloring Substance Chlorophyll from Leaves of Carica papaya.

Beautiful green leaves of Papaya are the rich source of Chlorophyll. Green color of chlorophyll has been used for a very long time as a natural colorant. Carica papaya has been considered as a good example and reasonable source of natural phytochemicals, which makes it suitable to color the food items and beverages. The aim of the present investigation is to develop the process of ultrasonic extraction in combination with solid phase extraction (SPE) to extract out chlorophyll with high yield as well as high degree of clarity. Newly customized ultrasonic-assisted extraction technique for the extraction of chlorophyll from Carica papaya leaves is optimized by taking different parameters like time, temperature, solvents concentrations, and raw material under consideration. Furthermore, the extract was purified by means of SPE and examined by using UV-Vis spectrophotometer. The highest yield of chlorophyll (dye) extract was found as 40% in solvent solution having 80 mL of ethanol and 20 mL of water with 5 minutes of extraction time, 35°C of temperature, and 1 grams of raw material in the sonication bath. Furthermore, the SPE purified sample was characterized by means of the UV-Vis spectrophotometer and here the total chlorophyll content was 34 mg/g, including chlorophyll a with a concentration of 14.1246 mg/g and chlorophyll b with concentration of 19.845 mg/g respectively. Consequently, sonication method can be suggested as a good method to get better concentration of chlorophyll.