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Taheri A, Bameri M, Sharifian S. Extraction of the phycobiliprotein pigments from the red seaweed Gelidium pusillum using an ultrasonic method combined with maceration or homogenization. pbp 2026; 8
URL: http://pbp.medilam.ac.ir/article-1-401-en.html
URL: http://pbp.medilam.ac.ir/article-1-401-en.html
1- Chabahar Maritime University, Faculty of Marine Sciences , taherienator@gmail.com
2- Chabahar Maritime University
3- Chabahar Maritime University, Faculty of Marine Sciences
2- Chabahar Maritime University
3- Chabahar Maritime University, Faculty of Marine Sciences
Abstract: (14 Views)
Objective: Red algae (Rhodophyta) are a great source of phycobiliproteins like R-phycoerythrin (R-PE) and R-phycocyanin (R-PC). These compounds have notable antioxidant, anti-inflammatory, and fluorescent qualities, making them useful in the food, pharmaceutical, and cosmetic sectors. Because of the resistant polysaccharide cell wall in species like Gelidium pusillum, extraction is not efficient. This study aimed to compare the efficiency of two combined extraction methods: ultrasound-assisted homogenization and ultrasound-assisted maceration, for the primary extraction of phycobiliproteins from G. pusillum collected from the Oman Sea, and to evaluate the antioxidant activity of the resulting extracts.
Methods: G. pusillum samples were collected during the fall from the Oman Sea in the Chabahar region and stored at -18°C. The thawed biomass was prepared for extraction by washing, manual fragmentation, and blending with a 0.1 M potassium phosphate buffer (pH 6.8) at a 1:15 (w/v) ratio. Using the ultrasound-maceration technique, the biomass underwent manual maceration and stirring in an ice bath for 90 minutes. Subsequently, it was sonicated at 20 kHz and 100% amplitude for 14 minutes in a pulse mode (9 seconds on, 1 second off), ensuring the temperature remained under 30°C. The ultrasound-homogenization approach involved initial mechanical homogenization of the biomass (20,000 rpm for 4 minutes) before identical ultrasonication. The supernatant was separated and kept after the centrifugation process (9000 rpm, 20 minutes, 4°C). Spectrophotometry was used to measure R-PE and R-PC levels at 564 and 618 nm. Purity ratios and extraction yields (mg/g fresh weight) were determined. The primary functional groups were identified using FTIR analysis. Antioxidant activity was measured using a DPPH radical scavenging assay within the 0.01–1 mg/mL concentration range, referencing BHT as a positive control. All experiments were carried out in triplicate and data were analyzed with t-tests or one-way ANOVA with Tukey's post-hoc test (significance at p < 0.05 and p < 0.01).
Results: The phycobiliprotein concentration obtained by the ultrasound-homogenization method was significantly higher than that of the ultrasound-maceration method. Total phycobiliprotein concentration was 0.0326 ± 0.003 mg/mL for homogenization and 0.0213 ± 0.002 mg/mL for maceration (p < 0.01). Similarly, the yield of total phycobiliproteins obtained by the homogenization method was 0.391 ± 0.029 mg/g fresh weight, which represented an increase of approximately 36% compared to the maceration method (0.288 ± 0.024 mg/g fresh weight; p < 0.01). R-PE and R-PC yields were also significantly higher in the homogenization method (0.184 ± 0.015 and 0.206 ± 0.012 mg/g, respectively) compared to maceration (0.129 ± 0.010 and 0.159 ± 0.012 mg/g; p < 0.01 for both). The FTIR analysis showed that two methods successfully extracted the phycobiliproteins, but the maceration extract contained some lipids. The DPPH radical scavenging activity of the extract from homogenization (0.5 mg/mL) measured 31.60 ± 4.82%, which was significantly higher than that of the extract from maceration (16.81 ± 6.07%). There was no significant difference at 1 mg/mL because of high variance in the maceration group. Both methods showed dose-dependent antioxidant activities with the highest scavenging at 1 mg/mL. Low purity ratios (from 0.0056 to 0.0145) were obtained for both methods, indicating polysaccharide contaminants and the need for additional purification steps. Total dry solid yields were similar for both methods (24.36 g/100 g wet weight for homogenization vs. 23.90 g/100 g for maceration) indicating comparable efficiencies in the extraction of total soluble compounds despite differences in the phycobiliprotein selectivity of the methods.
Conclusion: The results show that the ultrasound-assisted homogenization method is more efficient than the ultrasound-assisted maceration for the primary extraction of R-PE and R-PC from G. pusillum, leading to a 36% increase in total phycobiliprotein yield and improved antioxidant activity at moderate concentrations. The advantage of homogenization lies in its superior ability to disrupt the resistant polysaccharide cell wall of Gelidium, resulting in improved penetration and cavitation effects of ultrasound. The extracts showed preliminary antioxidant activity, which warrants further investigation, but industrial application is still premature until the extracts are purified and a safety evaluation is performed. Future work should be concentrated on RSM-based optimization, complementary antioxidant assays (ABTS, FRAP), encapsulation for stability, and structural verification by SEM/FTIR.
Methods: G. pusillum samples were collected during the fall from the Oman Sea in the Chabahar region and stored at -18°C. The thawed biomass was prepared for extraction by washing, manual fragmentation, and blending with a 0.1 M potassium phosphate buffer (pH 6.8) at a 1:15 (w/v) ratio. Using the ultrasound-maceration technique, the biomass underwent manual maceration and stirring in an ice bath for 90 minutes. Subsequently, it was sonicated at 20 kHz and 100% amplitude for 14 minutes in a pulse mode (9 seconds on, 1 second off), ensuring the temperature remained under 30°C. The ultrasound-homogenization approach involved initial mechanical homogenization of the biomass (20,000 rpm for 4 minutes) before identical ultrasonication. The supernatant was separated and kept after the centrifugation process (9000 rpm, 20 minutes, 4°C). Spectrophotometry was used to measure R-PE and R-PC levels at 564 and 618 nm. Purity ratios and extraction yields (mg/g fresh weight) were determined. The primary functional groups were identified using FTIR analysis. Antioxidant activity was measured using a DPPH radical scavenging assay within the 0.01–1 mg/mL concentration range, referencing BHT as a positive control. All experiments were carried out in triplicate and data were analyzed with t-tests or one-way ANOVA with Tukey's post-hoc test (significance at p < 0.05 and p < 0.01).
Results: The phycobiliprotein concentration obtained by the ultrasound-homogenization method was significantly higher than that of the ultrasound-maceration method. Total phycobiliprotein concentration was 0.0326 ± 0.003 mg/mL for homogenization and 0.0213 ± 0.002 mg/mL for maceration (p < 0.01). Similarly, the yield of total phycobiliproteins obtained by the homogenization method was 0.391 ± 0.029 mg/g fresh weight, which represented an increase of approximately 36% compared to the maceration method (0.288 ± 0.024 mg/g fresh weight; p < 0.01). R-PE and R-PC yields were also significantly higher in the homogenization method (0.184 ± 0.015 and 0.206 ± 0.012 mg/g, respectively) compared to maceration (0.129 ± 0.010 and 0.159 ± 0.012 mg/g; p < 0.01 for both). The FTIR analysis showed that two methods successfully extracted the phycobiliproteins, but the maceration extract contained some lipids. The DPPH radical scavenging activity of the extract from homogenization (0.5 mg/mL) measured 31.60 ± 4.82%, which was significantly higher than that of the extract from maceration (16.81 ± 6.07%). There was no significant difference at 1 mg/mL because of high variance in the maceration group. Both methods showed dose-dependent antioxidant activities with the highest scavenging at 1 mg/mL. Low purity ratios (from 0.0056 to 0.0145) were obtained for both methods, indicating polysaccharide contaminants and the need for additional purification steps. Total dry solid yields were similar for both methods (24.36 g/100 g wet weight for homogenization vs. 23.90 g/100 g for maceration) indicating comparable efficiencies in the extraction of total soluble compounds despite differences in the phycobiliprotein selectivity of the methods.
Conclusion: The results show that the ultrasound-assisted homogenization method is more efficient than the ultrasound-assisted maceration for the primary extraction of R-PE and R-PC from G. pusillum, leading to a 36% increase in total phycobiliprotein yield and improved antioxidant activity at moderate concentrations. The advantage of homogenization lies in its superior ability to disrupt the resistant polysaccharide cell wall of Gelidium, resulting in improved penetration and cavitation effects of ultrasound. The extracts showed preliminary antioxidant activity, which warrants further investigation, but industrial application is still premature until the extracts are purified and a safety evaluation is performed. Future work should be concentrated on RSM-based optimization, complementary antioxidant assays (ABTS, FRAP), encapsulation for stability, and structural verification by SEM/FTIR.
Keywords: Antioxidant activity, Phycoerythrin, Phycocyanin, Gelidium pusillum, Ultrasound-assisted extraction.
Type of Study: Research |
Subject:
Bioactive Compounds
Received: 2026/04/6 | Accepted: 2026/05/11 | Published: 2026/01/1
Received: 2026/04/6 | Accepted: 2026/05/11 | Published: 2026/01/1
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