Microwave-Assisted Extraction’s Kinetics of Phycobiliprotein from Spirulina Platensis: Influence of Citric Acid Concentration
DOI:
https://doi.org/10.48048/tis.2025.10042Keywords:
Phycobiliprotein, Citric acid, Peleg, Pseudo-first order, Pseudo-second orderAbstract
Phycobiliproteins (PBP), such as phycocyanin (PC), allophycocyanin (APC), and phycoerythrin (PE), are photosynthetic pigments in Spirulina platensis with antioxidant potential for food and medicine. This study optimized microwave-assisted extraction (MAE) of PBP with citric acid (1, 2, and 3 %) as a green solvent to enhance yield and stability using kinetic approach. MAE increased solvent temperature from 30 to 60 °C, accelerating PBP release while preserving its bioactivity. Three kinetic models, Peleg, pseudo-first order (PFO), and pseudo-second order (PSO), were applied to analyze extraction kinetics. Peleg effectively described the initial phase, indicating that higher CA concentrations enhanced PBP release. PFO captured the diffusion-driven phase, while PSO provided the most accurate equilibrium prediction by considering adsorption and solvent-protein interactions. Among the models, PSO showed the highest extraction capacity (Ce = 4.45 × 10⁻² mg/mL at 3 % CA), best R² (≥ 0.99), and lowest MAPE (3.45 %), confirming its superior fit at equilibrium. These findings highlight PSO as the best model for predicting maximum extraction, while Peleg and PFO remain relevant for the early and middle phases. The study confirms MAE with citric acid as an efficient, sustainable method for PBP extraction from Spirulina platensis.
HIGHLIGHTS
- MAE improves phycobiliprotein yield from Spirulina platensis.
- The addition of citric acid in solvent increases the extraction rate and capacity.
- Citric acid prevents phycobiliprotein degradation by controlling pH and oxidation.
- Kinetic models evaluate the influence of citric acid on extraction and stability.
- Peleg, PFO, and PSO models effectively describe the dynamics of PBP extraction.
GRAPHICAL ABSTRACT
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S Priyanka, R Varsha, R Verma and AS Babu. Spirulina: A spotlight on its nutraceutical properties and food processing applications. Journal of Microbiology, Biotechnology and Food Sciences 2023; 12(6), e4785.
BR Kiran and SV Mohan. Microalgal cell biofactory-therapeutic, nutraceutical and functional food applications. Plants 2021; 10(5), 836.
AH Khalil, EA Aidy, MA Said, R Kebeish and AH Al-Badwy. Biochemical and molecular docking-based assessment of Spirulina platensis’s bioactive constituents for their potential application as natural anticancer drug. Algal Research 2024; 82, 103624.
X Lv, Y Wu, M Gong, J Deng, Y Gu, Y Liu, J Li, G Du, R Ledesma-Amaro, L Liu and J Chen. Synthetic biology for future food: Research progress and future directions. Future Foods 2021; 3, 100025.
A Al-Farga, H Zhang, A Siddeeg, M Shamoon, MVM Chamba and N Al-Hajj. Proximate composition, functional properties, amino acid, mineral and vitamin contents of a novel food: Alhydwan (Boerhavia elegana Choisy) seed flour. Food Chemistry 2016; 211, 268-273.
KD Pamungkas, K Anam and T Budiati. Effect of high cell density to lipid content microalgae chlorella vulgaris on photoautotrophic cultivation. Food Science and Technology Journal 2023; 6(2), 78-85.
J Assuncao, HM Amaro, FX Malcata and AC Guedes. Factorial optimization of ultrasound-assisted extraction of phycocyanin from synechocystis salina: Towards a biorefinery approach. Life 2022; 12(9), 1389.
L Wils, C Leman-Loubiere, N Bellin, B Clement-Larosiere, M Pinault, S Chevalier, C Enguehard-Gueiffier, C Bodet and L Boudesocque-Delaye. Natural deep eutectic solvent formulations for spirulina: Preparation, intensification, and skin impact. Algal Research 2021; 56, 102317.
W Chen, J Xu, Q Yu, Z Yuan, X Kong, Y Sun, Z Wang, X Zhuang, Y Zhang and Y Guo. Structural insights reveal the effective Spirulina platensis cell wall dissociation methods for multi-output recovery. Bioresource Technology 2020; 300, 122628.
WF Elkot, A Elmahdy, TH El-Sawah, OA Alghamdia, SK Alhag, EA Al-Shahari, A Al-Farga, HA Ismail. Development and characterization of a novel flavored functional fermented whey-based sports beverage fortified with Spirulina platensis. International Journal of Biological Macromolecules 2024; 258(Part2), 128999.
T Lafarga, JM Fernandez-Sevilla, C Gonzalez-Lopez and FG Acien-Fernandez. Spirulina for the food and functional food industries. Food Research International 2020; 137, 109356.
N Hidhayati, KD Pamungkas, B Adinda and K Anam. Quality of arthrospira maxima in different water sources for cultivation media and biomass storage conditions. AIP Conference Proceedings 2023; 2972, 0500166.
Y Ladjal-Ettoumi, M Hamadi, LH Douik, Z Cherifi and A Nazir. Physicochemical, functional, and nutraceutical properties of spirulina and chlorella biomass: A comparative study. Algal Research 2024; 81, 103561.
X Li, W Hou, J Lei, H Chen and Q Wang. The unique light-harvesting system of the algal phycobilisome: Structure, assembly components, and functions. Prerpints.org, Basel, Switzerland, 2023.
H Chen, H Qi and P Xiong. Phycobiliproteins-a family of algae-derived biliproteins: Productions, characterization and pharmaceutical potentials. Marine Drugs 2022; 20(7), 450.
RDP Rodrigues, ASE Silva, TAV Carlos, AKP Bastos, RSD Santiago-Aguiar and MVP Rocha. Application of protic ionic liquids in the microwave-assisted extraction of phycobiliproteins from Arthrospira platensis with antioxidant activity. Separation and Purification Technology 2020; 252, 117448.
S Tavakoli, H Hong, K Wang, Q Yang, HH Gahruie, S Zhuang, Y Li, Y Liang, Y Tan and Y Luo. Ultrasonic-assisted food-grade solvent extraction of high-value added compounds from microalgae Spirulina platensis and evaluation of their antioxidant and antibacterial properties. Algal Research 2021; 60, 102493.
TJ Ashaolu. The powerful phycobiliproteins-phycocyanin and phycoerythrin: Pleiotropic applications and biofunctional uses. Algal Research 2024; 82, 103636.
K Limrujiwat, S Supan and W Khetkorn. Cyanobacterial biodiversity from Thai karstic caves as a potential source for phycobiliprotein production. Algal Research 2022; 64, 102666.
DY Susanti, WB Sediawan, M Fahrurrozi and M Hidayat. Optimization of agitation and kinetic studies on proanthocyanidin compound extraction from red sorghum grains in agitated vessel. IOP Conference Series: Materials Science and Engineering 2020; 778, 012085.
JM Santos, BC Jesus, H Ribeiro, A Martins, J Marto, M Fitas, P Pinto, C Alves, J Silva, R Pedrosa and IM Marrucho. Extraction of macroalgae phenolic compounds for cosmetic application using eutectic solvents. Algal Research 2024; 79, 103438.
E Ponthier, H Dominguez and MD Torres. The microwave assisted extraction sway on the features of antioxidant compounds and gelling biopolymers from Mastocarpus stellatus. Algal Research 2020; 51, 102081.
G Huschek, HM Rawel, T Schweikert, J Henkel-Oberlander and ST Sagu. Characterization and optimization of microwave-assisted extraction of B-phycoerythrin from Porphyridium purpureum using response surface methodology and Doehlert design. Bioresource Technology Reports 2022; 19, 101212.
S Zhou, W Chen and K Fan. Recent advances in combined ultrasound and microwave treatment for improving food processing efficiency and quality: A review. Food Bioscience 2024; 58(5), 103683.
V Darakai, C Punsawad, J Jitonnom, M Nisoa and P Rattanakit. Microwave-assisted ultrafine silver nanoparticle synthesis using Mitragyna speciosa for antimalarial applications. Green Processing and Synthesis 2024; 13(1), 20230257.
DA Esquivel-Hernandez, J Rodriguez-Rodriguez, M Rostro-Alanis, SP Cuellar-Bermudez, EI Mancera-Andrade, JE Nunez-Echevarria, JS Garcia-Perez, R Chandra, R Parra-Saldivar. Advancement of green process through microwave-assisted extraction of bioactive metabolites from Arthrospira Platensis and bioactivity evaluation. Bioresource Technology 2017; 224, 618-629.
N Orthesin, IT Hidayat, WT Wahyuni, UD Syafitri, Y Herbani and YW Sari. Optimization of chlorophyll extraction from dried spirulina platensis using low power microwave assisted extraction method. IOP Conference Series: Earth and Environmental Science 2024; 1359, 012021.
RN Fathimah, W Setyaningsih, C Carrera, AD Astari, RE Masithoh, IT Suryaningtyas and M Palma. A microwave-based technique to determine saccharides and polyols contents in Spirulina (Arthrospira platensis). Arabian Journal of Chemistry 2021; 14(4), 103094.
Y Liu, H Zhang, C Yi, K Quan and B Lin. Chemical composition, structure, physicochemical and functional properties of rice bran dietary fiber modified by cellulase treatment. Food Chemistry 2021; 342, 128352.
H Berrou, M Saleh and K Al-Ismail. Hydration kinetics of nixtamalized white bitter lupin (lupinus albus L.) seeds. Polish Journal of Food and Nutrition Sciences 2022; 72(4), 361-370.
X Cai, X Zhao, W Miao, Z Wu, HM Liu and X Wang. Optimization and kinetics modeling of microwave-assisted subcritical n-butane extraction of tigernut oil. Journal of Oleo Science 2022; 71(12), 1799-1811.
DY Susanti, WB Sediawan, M Fahrurrozi and M Hidayat. The effects of ultrasound wave on the extraction of proanthocyanidins from red sorghum grain using green solvent and a kinetics model of the extraction. Key Engineering Materials 2021; 884, 212-219.
DY Susanti, WB Sediawan, M Fahrurrozi and M Hidayat. A mechanistic model of mass transfer in the extraction of bioactive compounds from intact sorghum pericarp. Processes 2019; 7(11), 837.
DP Jaeschke, IR Teixeira, LDF Marczak and GD Mercali. Phycocyanin from spirulina: A review of extraction methods and stability. Food Research International 2021; 143, 110314.
A Adjali, I Clarot, Z Chen, E Marchioni and A Boudier. Physicochemical degradation of phycocyanin and means to improve its stability: A short review. Journal of Pharmaceutical Analysis 2022; 12(3), 406-414.
K Wang and G Luo. Microflow extraction: A review of recent development. Chemical Engineering Science 2017; 169, 18-33.
M Hassan, M Rahman, BC Ghos, I Hossain, A Amin and KA Zuhanee. Extraction, and characterization of CNC from waste sugarcane leaf sheath as a reinforcement of multifunctional bio-nanocomposite material: A waste to wealth approach. Carbon Trends 2024; 17, 100400.
AM Alomran and II Louki. Impact of irrigation systems on water saving and yield of greenhouse and open field cucumber production in Saudi Arabia. Agricultural Water Management 2024; 302(7), 108974.
JF Fabre, NUF Niangoran, C Gaignard, D Buso, Z Mouloungui and R Valentin. Extraction, purification and stability of C-phycocyanin from Arthrospira platensis. European Food Research and Technology 2022; 248, 1583-1599.
W Pan-utai, W Kahapana and S Iamtham. Extraction of C-phycocyanin from arthrospira (Spirulina) and its thermal stability with citric acid. Journal of Applied Phycology 2017; 30(1), 231-242.
B. Yuan, Z Li, H Shan, B Dashnyam, X Xu, DJ McClements, B Zhang, M Tan, Z Wang and C Cao. A review of recent strategies to improve the physical stability of phycocyanin. Current Research in Food Science 2022; 5, 2329-2337.
HF Izza, DY Susanti, S Mariyam and AD Saputro. Performance of microwave-assisted extraction of proanthocyanidins from red sorghum grain in various power and citric acid concentration. Journal of the Saudi Society of Agricultural Sciences 2023; 22(7), 480-492.
MM Hossain, R Ara, F Yasmin, M Suchi and W Zzaman. Microwave and ultrasound assisted extraction techniques with citric acid of pectin from Pomelo (Citrus maxima) peel. Measurement: Food 2024; 13, 100135.
W Routray and V Orsat. Variation of dielectric properties of aqueous solutions of ethanol and acids at various temperatures with low acid concentration levels. Physics and Chemistry of Liquids 2014; 52(2), 209-232.
Y Mao, J Robinson and E Binner. Understanding heat and mass transfer processes during microwave-assisted and conventional solvent extraction. Chemical Engineering Science 2021; 233, 116418.
V Lopez-Avila. Extraction with supercritical fluid inorganic extractions microwave-assisted extraction. Academic Press. California, United States, 2000.
I Efthymiopoulos, P Hellier, N Ladommatos, A Russo-Profili, A Eveleigh, A Aliev, A Kay and B Mills-Lamptey. Influence of solvent selection and extraction temperature on yield and composition of lipids extracted from spent coffee grounds. Industrial Crops and Products 2018; 119, 49-56.
DN Jamal, MSM Shajahan, N Cruzs, M Abhineshjayram and S Goutham. Empirical investigation and comparison of different viscosity liquids with increasing temperature. IEEE, New York, 2020.
X Liu, Y Tang, W Ning, Y Bao, T Luo and J Wang. Responses of issatchenkia terricola WJL-G4 upon citric acid stress. Molecules 2022; 27(9), 2664.
S Hussain, M Sharma, T Jarg, R Aav and R Bhat. Natural pigments (anthocyanins and chlorophyll) and antioxidants profiling of European red and green gooseberry (Ribes uva-crispa L.) extracted using green techniques (UAE-citric acid-mediated extraction). Current Research in Food Science 2023; 7, 100629.
VS Kislik. Principles of solvent extraction of organic and mineral acids. Elsevier Science, Amsterdam, Netherlands, 2012.
M Tang, H Zhang and L Li. Extraction remediation technologies of arsenic contaminated soils using citric acid. IEEE, New York, 2020.
J Wang, S Qin, J Lin, Q Wang, W Li and Y Gao. Phycobiliproteins from microalgae: Research progress in sustainable production and extraction processes. Biotechnology for Biofuels and Bioproducts 2023; 16, 170.
K Li, C Jiang, S Han, S Kang, J Chen, D Won, Y Kang, B Bae, Y Choi, HS Kim and J Lee. Green and efficient method to acquire high-value phycobiliprotein from microalgal biomass involving deep eutectic solvent-based ultrasound-assisted extraction. Food Chemistry 2024; 449, 139196.
Y Shmatok, H Potapenko and S Kirillov. Optimal design of LiMn2O4 for high-rate applications by means of citric acid aided route and microwave heating. Journal of Electrochemical Science and Engineering 2024; 14(3), 369-382.
M Jorns, S Strickland, M Mullins and D Pappas. Improved citric acid-derived carbon dots synthesis through microwave-based heating in a hydrothermal pressure vessel. RSC Advances 2022; 12(50), 32401-32414.
A Martinez, R Vargas and A Galano. Citric acid: A promising copper scavenger. Computational and Theoretical Chemistry 2018; 1133, 47-50.
YH Ju, SK Roy, AR Choudhury, S Kwon, J Choi, MA Rahman, T Katsube-Tanaka, T Shiraiwa, M Lee, K Cho and S Woo. Proteome changes reveal the protective roles of exogenous citric acid in alleviating cu toxicity in brassica napus L. International Journal of Molecular Sciences 2021; 22(11), 5879.
YC Shinta, B Zaman and S Sumiyati. Citric Acid and EDTA as chelating agents in phytoremediation of heavy metal in polluted soil: A review. IOP Conference Series: Earth and Environmental Science 2021; 896, 012023.
R Chaiklahan, N Chirasuwan and B Bunnag. Stability of phycocyanin extracted from Spirulina sp.: Influence of temperature, pH and preservatives. Process Biochemistry 2012; 47(4), 659-664.
VK Kannaujiya and RP Sinha. Thermokinetic stability of phycocyanin and phycoerythrin in food-grade preservatives. Journal of Applied Phycology 2016; 28(2), 1063-1070.
R Sharma, B Bhunia, A Mondal, TK Bandyopadhyay, I Devi, G Oinam, R Prasanna, G Abraham and ON Tiwari. Statistical optimization of process parameters for improvement of phycobiliproteins (PBPs) yield using ultrasound-assisted extraction and its kinetic study. Ultrasonics Sonochemistry 2020; 60, 104762.
F Wang, X Yu, Y Cui, L Xu, S Huo, Z Ding, Q Hu, W Xie, H Xiao and D Zhang. Efficient extraction of phycobiliproteins from dry biomass of Spirulina platensis using sodium chloride as extraction enhancer. Food Chemistry 2023; 406, 135005.
M Faieta, C Toong, MG Corradini, RD Ludescher and P Pittia. Degradation kinetics of C-Phycocyanin under isothermal and dynamic thermal treatments. Food Chemistry 2022; 382, 132266.
SM Zakaria, SMM Kamal, MR Harun, R Omar and SI Siajam. Extraction of phenolic compounds from Chlorella sp. microalgae using pressurized hot water: Kinetics study. Biomass Conversion and Biorefinery 2022; 12(1), 2081-2089.
OR Alara and NH Abdurahman. Microwave-assisted extraction of phenolics from Hibiscus sabdariffa calyces: Kinetic modelling and process intensification. Industrial Crops and Products 2019; 137, 528-535.
Hadiyanto, M Christwardana, H Sutanto, M Suzery, D Amelia and RF Aritonang. Kinetic study on the effects of sugar addition on the thermal degradation of phycocyanin from Spirulina sp. Food Bioscience 2018; 22, 85-90.
N Milicevic, P Kojic, M Sakac, A Misan, J Kojic, C Perussello, V Banjac, M Pojic and B Tiwari. Kinetic modelling of ultrasound-assisted extraction of phenolics from cereal brans. Ultrasonics Sonochemistry 2021; 79, 105761.
PR More and SS Arya. Intensification of bio-actives extraction from pomegranate peel via microwave irradiation: Effect of factors, optimization, kinetics, and bioactive profiling. Chemical Engineering and Processing - Process Intensification 2024; 202, 109839.
A Galetovic, F Seura, V Gallardo, R Graves, J Cortes, C Valdivia, J Nunez, C Tapia, I Neira, S Sanzana and B Gomez-Silva. Use of phycobiliproteins from atacama cyanobacteria as food colorants in a dairy beverage prototype. Foods 2020; 9(2), 244.
M Gomaa, SA Ali and AF Hifney. Enhancement of phycocyanin productivity and thermostability from Arthrospira platensis using organic acids. Microbial Cell Factories 2023; 22(1), 248.
B Nowruzi, M Ahmadi, N Bouaicha, AE Khajerahimi and SAA Anvar. Studying the impact of phycoerythrin on antioxidant and antimicrobial activity of the fresh rainbow trout fillets. Scientific Reports 2024; 14(1), 2470.
C Yang, D Lazidou, E Kampasakali, E Pavlidou and J Stratis. Mass transfer process of peanut protein extracted by bis(2-ethylhexyl) sodium sulfosuccinate reverse micelles. Grain and Oil Science and Technology 2024; 7(1), 60-67.
X Huang, J Wang, X Qu, C Huang and KL Yam. A release model considering chemical loss from a double-layer material into food. Thermal Science 2020; 24(4), 2419-2426.
A Delanney, A Ledoux and L Estel. Absorption in a pseudo-first-order regime with two irreversible reactions: Influence of the second reaction on the transfer rate. Chemical Engineering Journal 2024; 496, 153724.
P Zhou, X Li, J Zhou, Y Yang, J Zhi and L Shen. Mass transfer mechanism of the multivariate consecutive extraction process of pectin and hesperidin from Citrus aurantium L.: Kinetics, thermodynamics, diffusion and mass transfer coefficients. Separation and Purification Technology 2023; 311, 123339.
MG Fadl. Prediction of heavy metal biosorption mechanism through studying isotherm kinetic equations. Scientific Reports 2023; 13, 1576.
LAD Silva, MRDS Scapim, JFD Silva, AP Stafussa, ACR Aranha, LMDM Jorge, RO Defendi, ODOS Junior and GS Madrona. Modeling the extraction of bioactive compounds of green and red camu-camu peel and identification using UPLC-MS/MS. Chemical Engineering Research and Design 2023; 196, 1-12.
X Peng, P Yang, K Dai, Y Chen, X Chen, W Zhuang, H Ying and J Wu. Synthesis, adsorption and molecular simulation study of methylamine-modified hyper-cross-linked resins for efficient removal of citric acid from aqueous solution. Scientific Reports 2020; 10(1), 9623.
HN Tran. Differences between chemical reaction kinetics and adsorption kinetics: Fundamentals and discussion. Journal of Technical Education Science 2022; 70(1), 33-47.
L Gulua, T Khutsidze and T Turmanidze. Modeling of polyphenols extraction process from green tea and the effect of temperature on the process. International Journal of Science and Research Archive 2024; 12(1), 2115-2124.
Y Wang, C Wang, H Xue, Y Jin, M Yang and F Leng. Comparative analysis of three kinds of extraction kinetic models of crude polysaccharides from Codonopsis pilosula and evaluate the characteristics of crude polysaccharides. Biomass Conversion and Biorefinery 2023; 13, 12917-12933.
J Eilertsen, S Schnell and S Walcher. The michaelis–menten reaction at low substrate concentrations: Pseudo-first-order kinetics and conditions for timescale separation. Bulletin of Mathematical Biology 2024; 86, 68.
R Ezzati. A new insight into the pseudo-second-order model and the physical meaning of its rate constant in adsorption. Journal of Dispersion Science and Technology 2023; 46(2), 222-229.
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