Extraction Of Pigments Research Articles

Optimization of green extraction technologies for recovering bioactive compounds from Ixora coccinea waste flower biomass: A comparative response surface methodology and artificial neural network modeling

Red pigment from Ixora coccinea waste flower biomass was extracted using water as a cost-effective solvent for a sustainable source of organic color. The significant effect of selected variables on the method of extraction was studied for the green-extraction techniques, i.e., microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE), and conventional technique, i.e., heat-assisted extraction (HAE). Optimization of the pigment extraction was examined using response surface methodology (RSM) for each extraction technique. The parameters considered for MAE were power (W), time (s), pH, and solvent-to-sample ratio (mL/g) while temperature (°C), time (min), pH, and solvent-to-sample ratio (mL/g) were studied for both UAE and HAE. A training set for an artificial neural network (ANN) was created using the same design obtained from RSM. The modeling techniques (ANN and RSM) were then statistically differentiated by the coefficient of determination (R2), root mean square error (RMSE), and mean square error (MSE) which revealed the superiority of the RSM regression design over the ANN model. It was found that the energy consumption of pigment extraction was minimal for MAE. The obtained pigment was further characterized using Fourier transform infrared (FTIR) spectroscopy to reveal functional groups and a gas chromatography-mass spectrometry (GC-MS) to define its bioactive constituents.

An enzyme-fused phycobiliprotein synthesis system developed for visual whole-cell biosensors for the detection of cadmium during wastewater treatment

Transcription factor-based whole-cell biosensors are simple and inexpensive tools for monitoring heavy metals during environmental processes. However, most biosensors are depend on instrumentation or have low sensitivity. Here, all the enzymes in the phycobiliprotein synthesis system were fused with the phycocyanin apoprotein to create a system to synthesize enzyme-fused phycobiliproteins with comparable fluorescence. We replaced the biliprotein synthesis system T7 promoter with cadmium-sensing genetic elements to create two visual cadmium biosensors (Epcad-cbsA and Epcad-cbsS). Cd2+ was detected at nanomolar concentrations by measuring the fluorescence intensity and evaluating the degree of color change of the cell pellets. Moreover, good linear correlations were observed with Cd2+ concentrations up to 250nM. Compared with cellular fluorescence, our cadmium biosensor is more sensitive when cell color is used as the output signal, with detection limits of 7.6nM for Epcad-cbsA and 3.2nM for Epcad-cbsS. This visual platform was validated by assessing bioavailable cadmium during wastewater treatment. By immobilizing large amounts of phycobilin with the proteins in cells, our designed biosensors do not require pigment extraction or specialized light sources or detection devices. This work demonstrates that a biliprotein synthesis system can be a novel platform for the development of simple, low-cost, and highly sensitive visual biosensors for real-world applications.

Microbial Pigment Isolation, Culturing, and Extraction to Use as Textile Dyes

The textile industry is one of the largest worldwide polluters of clean water due to the heavy use of synthetic dyes. These chemicals negatively affect the environment, especially aquatic life due to their toxic and mutagenic properties. Synthetic dyes cause harm to human health such as skin allergies and respiratory sensitization. Several advantages such as ease of extraction, availability, high yields and no seasonal variation make microbial pigments the most ideal source of natural pigments. This study was done to isolate colour pigment producing bacteria and fungi from soil collected from organic farms from various locations in Sri Lanka. Out of 7 soil samples, 3 yielded pigment producing bacteria and fungi. In total, 9 pigment producing bacteria and 3 pigment producing fungi were isolated. Gause’s synthetic agar yielded the most pigmented isolates. Isolates were inoculated in broths and pigment production was observed. Extracellular pigments produced by 5 of the bacterial isolates were extracted by a water-based method. The antibacterial activity of the pigments in their crude and concentrated forms was tested using the well diffusion method against Escherichia coli ATCC 8739 and Staphylococcus aureus ATCC 6538P. Inhibition zone against S. aureus was observed for both crude (12.33±0.58mm) and concentrated pigments (9.67±0.58mm) extracted from purple pigment producing bacterial isolate (BPU). This pigment has the potential to be used in antibacterial textile preparation. Extracted pigments were used to dye scoured cotton fabric with the use of 3% alum as mordant. Pigment from BPU isolate resulted in better coloured fabric.

Unveiling the Intricacies of Microbial Pigments as Sustainable Alternatives to Synthetic Colorants: Recent Trends and Advancements

Bio-pigments are the colored primary and secondary metabolites released by microbes under stress conditions and are crucial for adaptation. Bio-pigments are being widely accepted for industrial utilization due to their natural form, organic source, and biodegradability. Also, the ease of cultivation, scalability and cost-effectiveness in terms of pigment extraction is bringing bio-pigments into the limelight. Chemical dyes are carcinogenic and pose a serious threat to human lives, which is another issue that environmentalists must address. However, bacterial pigments are safe to employ; therefore, the food, pharmaceutical, textile, and cosmetics sectors may all benefit from their applications. The therapeutic nature of bacterial pigments is revealed because of their antimicrobial, anticancer, cytotoxic, and remarkable antioxidant properties. Bio-pigments also have multifaceted properties and thus can be an attractive source for the next generation to live a sustainable life. The present review discusses the importance of bacterial pigments over synthetic dyes and their therapeutic and industrial potential. Extensive literature has been reviewed on the biomedical application of bacterial pigments, and further opportunities and future challenges have been discussed.

The Profile of Phenolic Compounds Identified in Pitaya Fruits, Health Effects, and Food Applications: An Integrative Review.

This integrative review aimed to identify the phenolic compounds present in pitayas (dragon fruit). We employed a comprehensive search strategy, encompassing full-text articles published between 2013 and 2023 in Portuguese, English, and Spanish from databases indexed in ScienceDirect, Capes Periodics, Scielo, and PubMed. The study's selection was guided by the question, "What are the main phenolic compounds found in pitaya fruits?". After screening 601 papers, 57 met the inclusion criteria. The identified phytochemicals have been associated with a range of health benefits, including antioxidant, anti-inflammatory, and anxiolytic properties. Additionally, they exhibit promising applications in the management of cancer, diabetes, and obesity. These 57 studies encompassed various genera, including Hylocereus, Selenicereus, and Stenocereus. Notably, Hylocereus undatus and Hylocereus polyrhizus emerged as the most extensively characterized species regarding polyphenol content. Analysis revealed that flavonoids, particularly kaempferol and rutin, were the predominant phenolic class within the pulp and peel of these fruits. Additionally, hydroxycinnamic and benzoic acid derivatives, especially chlorogenic acid, caffeic, protocatechuic, synaptic, and ellagic acid, were frequently reported. Furthermore, betalains, specifically betacyanins, were identified, contributing to the characteristic purplish-red color of the pitaya peel and pulp. These betalains hold significant potential as natural colorants in the food industry. Therefore, the different pitayas have promising sources for the extraction of pigments for incorporation in the food industry. We recommend further studies investigate their potential as nutraceuticals.

Microbial synthesis of zinc oxide nanoparticles and their potential biological application as an antimicrobial and anticancer agent

Zinc oxide nanoparticles (ZnO-NPs) are a type of metal oxide nanomaterial, recognized as a valuable and adaptable inorganic compound owing to its distinctive physical and chemical properties. Nanosized ZnO particles exhibit substantial antibacterial properties attributable to their diminutive size, which can activate various bactericidal mechanisms within the bacterial cell, including interactions with the bacterial surface or core, the generation of reactive oxygen species (ROS), the release of Zn2+, and potential endocytosis by cells. ZnO NPs nanoparticles were extracellularly produced using pigment extracts from the PP6 strain. Agar well screening indicated that PP6 secondary metabolites possess antibacterial properties. UV–Vis spectroscopy was used to analyze the external growth of nanoparticles. Scanning Electron Microcopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), and X-ray diffraction (XRD) techniques were employed to analyze the shape, stability, crystalline structure, and coating of strain PP6 ZnO NPs. The PP6 ZnO NPs demonstrated an antibiofilm impact on the bacterial pathogens tested, which was dependent on the dosage. Elevated levels of lipid peroxidation (LPO) and reduced antioxidant activity are indicative of apoptosis in cancer cells. The synthesized ZnO NPs nanoparticles were assessed for their anticancer properties by performing the MTT assay on HT-29 human colorectal adenocarcinoma cells. ZnO NPs nanoparticles exposed to HT-29 cells the viability was reduced significantly in proportion to the concentration of nanoparticles. Additional comprehensive study will be needed to fully understand their mechanism.

Bio-prospecting the Anti-oxidative and Radioprotective Role of Bioactive Pigment Isolated from Pontibacter indicus

Aim: To evaluate the anti-oxidative and radioprotective role of Pontibacter indicus SCG24 pigment during radiation exposure Background: Radiation-induced cytotoxicity is quite common during cancer therapy. There is a need for naturally derived therapeutic molecules that can scavenge free radicals. They may act as substitutes for synthetic molecules. Hence, there is a need for urgent evaluation of these potent compounds before therapeutic application. Objective: The objective of this study is to examine the anti-oxidative and radioprotective role of P. indicus SCG24 pigment, specifically to evaluate free radical scavenging X-ray irradiated HDF cells Method: A radiotoleraent pigment-producing P. indicus SCG24 was isolated from pharmaceutical effluent. Chloroform was used as a primary solvent for pigment extraction. GCMS/MS analysed initial pigment composition. Various In-vitro antioxidant assays were performed using ABTS, FRAP, and DPPH assay. Flow cytometry was used to determine the rate of scavenging activity of pigment in HDF cells. Results: The GCMS/MS profile of the chloroform extract revealed twenty-two compounds. Furthermore, based on the DPPH, ABTS, and FRAP assay, the pigment was found to have significant antioxidant properties. The flow cytometry results indicate that the pigment possesses radioprotectant activity by neutralizing ROS species in HDF cells when exposed to X-ray radiation. Conclusion: These observations on P. indicus SCG24 pigment suggested that the pigment may have potential therapeutic importance.

Exploration of the Bioactivity of Pigmented Extracts from Streptomyces Strains Isolated Along the Banks of the Guaviare and Arauca Rivers (Colombia)

Pigments are chemical compounds that impart color through mechanisms such as absorption, reflection, and refraction. While traditional natural pigments are derived from plant and insect tissues, microorganisms, including bacteria, yeasts, algae, and filamentous fungi, have emerged as promising sources for pigment production. In this study, we focused on pigment production by 20 Streptomyces isolates from our in-house actinobacteria strain collection, sourced from the Guaviare and Arauca Rivers in Colombia. The isolates were identified via 16S rRNA gene sequencing, and the bioactivities—including antioxidant, antibacterial, and cytotoxic properties—of their extracts obtained across four different culture media were assessed. Promising pigmented hydroalcoholic extracts demonstrating these bioactivities were further analyzed using LC-MS, leading to the annotation of a variety of pigment-related compounds. This study revealed that culture media significantly influenced both pigment production and bioactivity outcomes. Notably, anthraquinones, phenazines, and naphthoquinones were predominant pigment classes associated with cytotoxic and antimicrobial activities, while carotenoids were linked to antioxidant effects. For instance, S. murinus 4C171 produced various compounds exhibiting both cytotoxic and antioxidant activities. These findings highlighted a growth medium-dependent effect, as pigment production, coloration, and bioactivity outcomes were influenced by growth media. These results demonstrate the significant potential of Streptomyces isolates as sources of bioactive pigments for diverse applications.

Studying the Stability of Anthocyanin Pigments Isolated from Juices of Colored-Fleshed Potatoes

The aim of this study was to obtain extracts of anthocyanin pigments from red and purple-fleshed potato juices characterized by stable color. For this purpose, potato juices were pasteurized at different temperatures or fruit and vegetable concentrates were added to them. Color stability tests of the obtained pigments were carried out in model pH and temperature conditions and after adding to natural yogurt. Both the pasteurization process and the addition of fruit and vegetable concentrates to the potato juices positively affected their color and its stability in time. However, the pasteurization of the potato juices had a negative effect on the content of biologically active compounds, in contrast to the juices stabilized with the addition of fruit and vegetable concentrates. Anthocyanin pigments from red-fleshed potato juices were more stable than those isolated from the purple-fleshed potato juices. The results of model tests of the anthocyanin pigment concentrates from the colored-flesh potatoes and natural yoghurts with their addition confirmed the high stability of the tested concentrates.

Extraction of dye sorghum biocolorant for the dyeing of wagashi, a West African soft cheese

We developed a sustainable method to extract the red biocolorant from dye sorghum leaf sheaths for dyeing wagashi, a West African soft cheese. The pigments were extracted using three different solvents: commercial ethanol (method A), sodabi (a local liquor distilled from palm wine; method B) and aqueous alkaline solvent (method C). In methods A and B, a pot still was used to distil and collect the solvent for reuse. The obtained extracts were used to dye wagashi at various concentrations. The yield and colour characteristics of the extracts were evaluated and the physicochemical characteristics of the dyed wagashi were recorded and compared to commercial wagashi sold in the local market. The solvent recovery in methods A and B was 96 % and 47 %, respectively. Extraction of anthocyanin pigments was highest with method A, namely 21 mg mL−1 for total phenolics against 13.4 and 1.64 mg mL−1 for methods B and C, respectively. The colour parameters suggested a similar colour for all the extracts, but the redness index of the extract from method B was significantly higher than that of the other methods. Wagashi dyed with extracts from methods B and C had similar colour characteristics and were comparable to the best-coloured commercial wagashi from the Parakou market based on the chroma and the hue angle values of the chromaticity diagram. Despite the lower colouring power of extracts made by method B, we consider this method the best when taking into account the cost and availability of the solvent. In conclusion, the application of sodabi extract at a dose of 200 mL L−1 is recommended to dye wagashi.

Extraction and separation of pigments from Saccharina latissima using eutectic solvents

Harnessing the untapped potential of marine biomass has emerged as a pivotal strategy to address growing demands for natural bioactive compounds across many industries. Among these, Saccharina latissima, a prolific marine alga, is a rich source of fucoxanthin and chlorophylls — molecules of significant biotechnological interest. This work used Saccharina latissima as a source of pigments to develop an integrated platform to promote the extraction and separation of chlorophyll and fucoxanthin using eutectic solvents (ES). Hydrophobic ES were investigated in the extraction of these pigments, and operational conditions were optimised. Among four hydrophobic ES screened, the Ment:LevA system, enhanced with 20 % (v/v) of water, exhibited the greatest potential, yielding an optimised extract rich in fucoxanthin with 137.2 ± 2.6 µgfucoxanthin·gbiomass-1. Additionally, a unique ES-ES biphasic system facilitated the selective partitioning of pigments: chlorophylls predominantly remained in the hydrophobic phase, while 95 % of fucoxanthin migrated to the hydrophilic. To further refine the quality of the extracted fucoxanthin, a subsequent purification step using water was implemented successfully, resulting in a concentrated pigment product. This work highlights ES as potential biocompatible solvents for the recovery of value-added compounds from marine biomass.

Fabrication of dye-sensitized solar cells with natural dye pigments derived from mustard green (Brassica juncea L) and turmeric (Curcuma longa L)

Abstract Natural dye molecules are good alternatives for the ruthenium-based sensitizer for use in dye-sensitized solar cell (DSSC) application. In this research, we have reported the potential application of natural pigment extracts as a dye-sensitizer in DSSC. Natural dyes were extracted using simple maceration technique from natural sources such as mustard green (Brassica juncea L) and turmeric (Curcuma longa L). UV-Vis and FT-IR characterization were employed to examined the optical characteristics and identify their functional groups of the dyes, respectively. The UV-Vis characterization of mustard green dye and turmeric dye show a wide absorption in visible region. The FT-IR spectrum of mustard green dye and turmeric dye show the presence the anchoring groups. The DSSC’s performance was studied using current-voltage (I-V) measurements and electrochemical impedance spectroscopy (EIS). It was observed that the DSSC with turmeric dye shows better photovoltaic performance than DSSC with mustard green dye correlated to its longer charge carrier lifetime.

Extraction of Bacterial Pigments and Evaluation of its Antioxidant Activity

Carotenoid are pigment that exist in a wide variety of plant and microorganism they are characterized by red, yellow or orange coloration .These important pigment play significant role in animal and human health .In contrast to plant production, the advantages of microbial fermentation of carotenoids were lower media cost, fast growth rate of microorganism and the ease of culture control . When the microbial cells are used to produce the color the term refers to ‘microbial pigments’. Different types of pigments extracted from different microbes. In this study, soil and water samples were collected from different areas of MGM University, Chh. Sambhajinagar .Different pigmented colonies were observed on nutrient agar plate .Out of these, the orange and yellow colonies were selected and sub-cultured further to obtain their pure cultures. These samples were inoculated onto nutrient agar plates, and after an incubation period of 24 hours, different colonies were observed on the plates. Out of these, the orange and yellow isolated were selected and sub-cultured further to obtain their pure cultures. After production of the pigment in liquid medium, it was further processed for the extraction of these pigments. Further, antioxidant assay by radical scavenging activity of DPPH was carried out, which revealed that the pigment had an radical scavenging activity of 60% as compared to that of the standard ascorbic acid, which showed radical scavenging activity of 33 %. Thus, the current study can be a useful step for large-scale production of pigments, their purification and application in cosmetic industries

EXPLOITING ECO-FRIENDLY NATURAL DYES FROM PLANT SOURCES: EXTRACTION AND DIVERSE APPLICATIONS

Natural pigments extracted from plant sources offer eco-friendly and sustainable alternatives to synthetic colorants. In this study, pigment samples were obtained through aqueous extraction from beetroot (S1), red cabbage (S2), and turmeric (S3). Their physicochemical properties and potential applications were thoroughly evaluated. The research problem addressed in this study is the need to overcome the limitations of natural dyes, such a slower color fastness, reduced reproducibility, and limited color range, in order to promote their wider adoption in various industries as sustainable alternatives to synthetic colorants. The pigment extracts were assessed for their suitability in textile dyeing, paper pH indicators, food coloring, ink production, and natural stamp pad formulations. Cotton and wool fabrics were dyed by adding traditional mordants, and the pigment-treated textiles exhibited distinctive color changes and varying levels of color fastness. The pigment samples demonstrated the ability to act as effective acid-base indicators, displaying characteristic color transitions in response to varying pH levels. When used as natural food colorants, the extracts imparted attractive hues to rice and cream without altering their flavor profiles. The feasibility of incorporating these plant-derived pigments in ink manufacturing and stamp pad production was also explored. Beetroot pigment exhibited superior color retention and fastness properties, while red cabbage and turmeric extracts displayed effectiveness in selected applications. The findings of this comprehensive study highlight the viability of these plant sources as natural substitutes for synthetic colorants in diverse fields. The observed performance characteristics and the eco-friendly nature of the pigments underscore their potential to serve as sustainable alternatives in various industries.

Hydrogen-rich solvent method enhances the extraction of phenolics, pigments, reducing sugars, organic acids, and vitamin C from cowslip (Primula veris L.) flower

Cowslip (Primula veris L.) is an anioxidant-rich plant used for many food and medicinal purposes. In this study, the effect of incorporating hydrogen (H2) into water (HRW), ethanol (HRE), and methanol (HRM) on the extraction of flavonoids (TFC), phenolics (TPC), and antioxidants (metal chelation, FRAP, DPPH, and ABTS) as well as color (L*, a*, b*, C*, H°), chlorophyll (a, b), lycopene, and β-carotene of wild cowslip flowers was evaluated. The results were submitted to Principal Component Analysis and correlation. Phenolic compounds, organic acids, vitamin C, and sugar content of the extract were analyzed using high-performance reverse phase liquid chromatography (HPLC-DAD/RID). The highest TPC, TFC, and antioxidant activity (metal chelation, FRAP, DPPH, and ABTS) were shown for HRM samples, followed by HRE samples, while the lowest were found for pure solvents. Incorporating H2 into all solvents significantly increased TPC, TFC, metal chelation, DPPH, and ABTS scavenging activity, as well as b*, C*, H°, chlorophyll, lycopene, and β-carotene values. The levels of some phenolics (catechin, epicatechin, rutin, and quercetin), organic acids (tartaric acid and succinic acid), reducing sugars (maltose, glucose, and fructose), and ascorbic acid increased by 10–90 % when an H2-rich solvent was used instead of the pure solvent. This is the first report showing the enhanced extraction of organic acids, pigments, and ascorbic acid by hydrogen-rich solvent. The main benefits of the HRS method are its sustainability and high efficiency in extracting bioactive compounds from medicinal plants.

Extraction of polyphenols from Apocynum venetum leaves using customized deep eutectic solvents: Process optimization and antioxidant evaluation

To encourage the commercial utilization of A. venetum polyphenols as bio-colorant on textile, five different combinations of deep eutectic solvents (DESs) were evaluated using a single-factor approach and the extraction of polyphenols from A. venetum leaves by ultrasound-assisted DESs was optimized. At the same time, the antioxidant activity of polyphenols prepared were investigated. The DESs composed of choline chloride and glycerol with a molar ratio of 1:2 and a water content of 20 % showed higher extraction performance than other DESs and traditional solvents (water, 60 % ethanol and 60 % methanol). With the most effective solvent the maximum yield 8.24 % of polyphenols was obtained under the optimal extraction conditions were as follows: the ratio of liquid volume (mL) to solid mass (g) is 49:1, extraction temperature of 77 ℃, and extraction time of 55 min. In addition, the polyphenols that have been separated and purified from the DESs extract exhibited antioxidant activities in three kinds of assays including 2,2-diphenyl-1-picrylhydrazyl stable free radical (DPPH), 2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid free radicals (ABTS) and hydroxyl radical (•OH) scavenging activity. Therefore, this paper demonstrates that DESs are effective and environmentally friendly solvents extraction of antioxidant polyphenols pigments from the leaves of A. venetum.

Optimal extraction, phytochemical and electrochemical potential assessment of pigments from Persicaria Lapathifolia for dye-sensitized solar cell application

In this study, we concentrated on optimal extraction, phytochemical, and electrochemical assessment of natural pigments from the roots and flowers of Persicaria lapathifolia as new sensitizers in dye-sensitized solar cells. The extraction technique required optimizing the use of acidified ethanol with acetic acid and hydrochloric acid as flower and root extractants, respectively. Photo-electrochemical tests revealed that these pigments could improve the efficiency of DSSCS. The optical characterization revealed maximal absorbance peaks at 462 and 654 nm for root dye and 535, 606 and 666 nm for blossom dye. Phytochemical examination revealed the presence of anthocyanins flavonoids glycoside linkage and betacyanin in both extracts. The Cyclic voltammetry tests revealed oxidation potential peaks in root and floral dyes the HOMO and LUMO potentials were calculated for both dyes, revealing values that could help improve DSSC performance. The DSSCS sensitized with flower extract had an open circuit voltage of 0.649 V a brief current pulse of 3.8 mA and an efficiency of 1.6335%. On the other hand root, pigment-based DSSCS performed slightly inferior with a Voc of 0.55 V an Isc of 3.7 mA, and an efficiency of 1.33%. These findings point to the potential of Persicaria lapathifolia pigments to boost DSSC efficiency emphasizing the relevance of sustainable energy sources. More research and optimization are required to realize the potential of these natural pigments for DSSC applications. This study adds to the investigation of green energy options, notably rooftop photovoltaic systems, in response to an increasing desire for ecologically sound energy solutions.

Extraction of Anthocyanin Pigments from the Peel of Dragon Fruit for Food Coloring

Extraction of Anthocyanin Pigments from the Peel of Dragon Fruit for Food Coloring

Color Fastness and Damage of Hair Dyed with Aplysia kurodai Pigment Extracts

Color Fastness and Damage of Hair Dyed with Aplysia kurodai Pigment Extracts

Optimization and extraction of annatto pigments obtained from Bixa orellana L. using supercritical fluid extraction

Traditionally, various techniques like water, vegetable oil, and solvent extraction are used to extract annatto pigments, a natural coloring agent derived from the Bixa orellana L. shrub. These traditional extraction methods can be time consuming and yield minimal pigment concentrations. This study explores an innovative approach to extract the maximum pigments from the annatto seed using supercritical fluid extraction (SFE). Fresh, extracted aqueous, and peeled samples of annatto powder were used at different temperatures (40, 45, and 50 °C) using different solvents of ethanol, methanol, and ethyl acetate with 99 % purity. The optimal extraction condition in supercritical fluid extraction is obtained with an aqueous sample treated with methanol solvent at 45 °C that yields 4.62 % norbixin, 2.49 % bixin, and a hue angle of 24.85⁰, achieving a high desirability value (0.949). Response surface method (RSM) was used to optimize the extraction procedure to obtain a higher yield of annatto pigments. In addition, for optimized conditions, the total phenols and antioxidants assay yielded 3.8 mg GAE/g and 51 % inhibition. HPLC analysis confirmed norbixin in the extracts obtained with a retention time of 38.40 min. This method offers a significant boost in pigment extraction efficiency, but also minimizes processing time. These combined advantages position it as a potentially superior approach for producing annatto pigments with enhanced concentration and purity.