Vietnam Academy of Science and Technology

Project's information

Project's title	Study of the process of isolating natural toxins from marine organisms
Project’s code	TĐĐTB0.06/21-23
Research hosting institution	Institute of Chemistry
Project leader’s name	Dr. Tran Dang Thuan
Project duration	01/01/2021 - 31/12/2023
Project’s budget	6,000 million VND
Classify	Grade A
Goal and objectives of the project	- Development of protocols for isolation of natural toxins from marine organisms; - Purification of standard toxins to satisfactory purity for analysis and testing.
Main results	- Theoretical results: + TTX molecule imprinted magnetic nanomaterials (Fe3O4@SiO2-TTXMIP) were successfully synthesized. The adsorption capability of Fe3O4@SiO2-TTXMIP materials towards TTX molecule and desorption capability of TTX molecule from Fe3O4@SiO2-TTXMIP-TTX materials were investigated and optimized. TTX toxin adsorption efficiency of Fe3O4@SiO2-TTXMIP reached 96.5% under optimal conditions of the initial TTX standard concentration of 10,000 μg/L, adsorbent dosage of 8 g/L, pH of 4-5, stirring rate of 150 rpm, temperature of 35-40 oC and adsorption time of 60 min. TTX toxin desorption efficiency from Fe3O4@SiO2-TTXMIP-TTX reached 99% in AA/ACN solvent (1%, v/v) (8 g/L) under temperature condition of 50 oC, stirring rate of 150 rpm and adsorption time 50 min. + STX molecule imprinted magnetic nanomaterials (Fe3O4@SiO2-STXMIP) were successfully synthesized. The adsorption capability of Fe3O4@SiO2-STXMIP materials towards STX molecule and desorption capability of STX molecule from Fe3O4@SiO2-STXMIP-STX materials were investigated and optimized. STX toxin adsorption efficiency of Fe3O4@SiO2-STXMIP reached 93.5% under optimal conditions of the initial STX standard concentration of 1000-1500 µg/L, adsorbent dosage of 4 g/L, pH of 9, stirring rate of 150 rpm, temperature of 35 oC and adsorption time of 150 min. STX toxin desorption efficiency from Fe3O4@SiO2-STXMIP-STX reached 99.5% in AA/ACN solvent (0,1%, v/v) (4 g/L) under temperature condition of 40 oC, stirring rate of 150 rpm and adsorption time 90 min. + Materials including Fe3O4, Fe3O4@SiO2, Fe3O4@SiO2-TTXMIP, Fe3O4@SiO2-TTXMIP-TTX, Fe3O4@SiO2-STXMIP, và Fe3O4@SiO2-STXMIP-STX were characterized with TEM, SEM, VSM, XRD, TGA, EDS and FT-IR. Data revealed that the studied materials exhibited magnetization level of over 20 emu/g, which is good enough for separatation and recovery of the materials using magnetic devices. Surface area of these materials were determined in the range of 56-127 m2/g. Crystall structures of the studied materials e.g., Fe3O4@SiO2-TTXMIP, Fe3O4@SiO2-TTXMIP-TTX, Fe3O4@SiO2-STXMIP, và Fe3O4@SiO2-STXMIP-STX were remained as structures of Fe3O4 và Fe3O4@SiO2 during the synthesis process, demonstrating the excellent stability of the targeted materials. Analysis of functional and chemical elements of the materials showed that TTX and STX were sucessfully adsorbed on Fe3O4@SiO2-TTXMIP and Fe3O4@SiO2-STXMIP and desorped from Fe3O4@SiO2-TTXMIP-TTX and Fe3O4@SiO2-STXMIP-STX, respectively. Applied results: + A protocol of synthesis of TTX molecule imprinted magnetic nanomaterials (Fe3O4@SiO2-TTXMIP) at the scale of 87.5 g/batch was sucessfully developed; + A protocol of enrichment of TTX from pufferfish viscera to obtain the pufferfish viscera extract containing total TTXs concentration of 9,46 mg/L (TTX = 81.6%) at the scale of 5 kg material/batch was sucessfully developed; + A protocol of purification of TTX from the TTX-enriched pufferfish viscera extract (TTXs = 9.46 mg/L, TTX = 81.6%) to the purified TTX of 95.7% purity using Fe3O4@SiO2-TTXMIP at the scale of 6.85 mg/batch was sucessfully developed; + The amount of 2 mg purified TTX with purity of 95.7% was determined by LC-MS/MS; + A protocol of synthesis of STX molecule imprinted magnetic nanomaterials (Fe3O4@SiO2-STXMIP) at the scale of 91.2 g/batch was sucessfully developed; + A protocol of enrichment of STX from Carcinoscorpius rotundicauda’s soft tissues to obtain the Carcinoscorpius rotundicauda extract containing total STXs concentration of 1222 µg/L (STX = 90.7%) at the scale of 25 kg material/batch was sucessfully developed; + A protocol of purification of STX from the STX-enriched Carcinoscorpius rotundicauda’s (STXs = 1222 µg/L, STX = 90.7%) to the purified STX of 95% purity using Fe3O4@SiO2-STXMIP at the scale of 4.77 mg/batch was sucessfully developed; + The amount of 2 mg purified STX with purity of 95% was determined by 1H-NMR.
Novelty and actuality and scientific meaningfulness of the results	+ A protocol of synthesis of TTX molecule imprinted magnetic nanomaterials (Fe3O4@SiO2-TTXMIP) at the scale of 87.5 g/batch was sucessfully developed; + A protocol of enrichment of TTX from pufferfish viscera to obtain the pufferfish viscera extract containing total TTXs concentration of 9,46 mg/L (TTX = 81.6%) at the scale of 5 kg material/batch was sucessfully developed; + A protocol of purification of TTX from the TTX-enriched pufferfish viscera extract (TTXs = 9.46 mg/L, TTX = 81.6%) to the purified TTX of 95.7% purity using Fe3O4@SiO2-TTXMIP at the scale of 6.85 mg/batch was sucessfully developed; + The amount of 2 mg purified TTX with purity of 95.7% was determined by LC-MS/MS; + A protocol of synthesis of STX molecule imprinted magnetic nanomaterials (Fe3O4@SiO2-STXMIP) at the scale of 91.2 g/batch was sucessfully developed; + A protocol of enrichment of STX from Carcinoscorpius rotundicauda’s soft tissues to obtain the Carcinoscorpius rotundicauda extract containing total STXs concentration of 1222 µg/L (STX = 90.7%) at the scale of 25 kg material/batch was sucessfully developed; + A protocol of purification of STX from the STX-enriched Carcinoscorpius rotundicauda’s (STXs = 1222 µg/L, STX = 90.7%) to the purified STX of 95% purity using Fe3O4@SiO2-STXMIP at the scale of 4.77 mg/batch was sucessfully developed; + The amount of 2 mg purified STX with purity of 95% was determined by 1H-NMR.
Products of the project	- Scientific papers in referred journals (list): 1) Dang Thuan Tran, Cam Van T. Do, Cuc T. Dinh, Mai T. Dang, Khanh Hy Le Ho, Truong Giang Le and Viet Ha Dao (2023). Recovery of tetrodotoxin from pufferfish viscera extract by amine-functionalized magnetic nanocomposites, RSC Advances, 13, 18108–18121 (SCI-E, IF= 4.036, Q2); 2) Dang Thuan Tran, Cuc T. Dinh, Mai T. Dang, Cam Van T. Do, Khanh Hy Le Ho, Truong Giang Le, Viet Ha Dao (2023). Central composite design-based response surface methodology for optimization of tetrodotoxin extraction from the horseshoe crab Carcinoscorpius rotundicauda. Asia-Pac Journal of Chemical Engineering, e2974. doi:10.1002/apj.2974 (SCI-E, IF= 1.8, Q3). doi:10.1002/apj.2974; 3) Do Thi Cam Van, Nguyen Thi Phuong Dung, Tran Dang Thuan,* (2023). Phát triển qui trình chiết xuất nâng cấp để thu hồi tetrodotoxin từ nội tạng cá nóc. Tạp chí Khoa học & Công nghệ Trường Đại học Công nghiệp Hà Nội, Vol. 59 - No. 6B, trang 128-134. http://hdl.handle.net/123456789/2741; 4) Dinh Thi Cuc, Dang Thi Mai, Do Thi Cam Van, Tran Huu Quang, Tran Dang Thuan* (2023). Seasonal, body weight and gender variation of tetrodotoxin in horseshoe crab Carcinoscorpius rotundicauda collected from Can Gio coast, Ho Chi Minh city, Vietnam. Tạp chí Khoa học Trường Đại học Quy Nhơn, 7(5), 61-73. DOI: https://doi.org/10.52111/qnjs.2023.17506. - Patents (list): 1) 01 Invention: PHƯƠNG PHÁP CHẾ TẠO VẬT LIỆU NANO TỪ TÍNH IN DẤU PHÂN TỬ TETRODOTOXIN (accepted a valid application by the National Office of Intellectual Property on 29/08/2023) 2) 01 Invention: QUY TRÌNH TINH CHẾ TETRODOTOXIN TỪ NỘI TẠNG CÁ NÓC DÙNG VẬT LIỆU NANO TỪ TÍNH IN DẤU PHÂN TỬ TETRODOTOXIN (accepted a valid application by the National Office of Intellectual Property on 29/08/2023) - Technological products (describe in details: technical characteristics, place): 1) 01 protocol for purification of TTX from pufferfish viscera TTXs-rich extract using Fe3O4@SiO2-TTXMIP to TTX purity of 95.7%; 2) 2 mg pure TTX (purity of 95.7%), stored at Laboratory of Biochemical Technology, Institute of Chemistry - Vietnam Academy of Science and Technology; 3) 01 protocol for purification of STX from Carcinoscorpius rotundicauda STXs-rich extract using Fe3O4@SiO2-STXMIP to STX purity of 95.0%; 4) 2 mg pure STX (purity of 95.0%), stored at Laboratory of Biochemical Technology, Institute of Chemistry - Vietnam Academy of Science and Technology; - Other products (if applicable): 01 Master student
Research region	The scientific-value results of the project will be referencial materials for institutes and research teams in Vietnam Academy of Science and Technology carrying out the similar research direction.
Recommendations	The research team kindly suggest that Vietnam Academy of Science and Technology continues to further provide financial support for our team conducting comprehensive and intensive researches in this direction to produce larger quantity of pure toxins.
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