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標題: 以鹽輔助均相液液微萃取技術結合HPLC-UV快速偵測微生物發酵培養液中的表面素
Rapid Determination of Surfactin in Microbial Fermentation Broths using Salt-Assisted Homogeneous Liquid-Liquid Microextraction Technique combined with HPLC-UV
作者: 杜怡真
Du, Yi-Chen
關鍵字: Surfactin;表面素;Microbial fermentation broth samples;Salt-assisted homogeneous liquid-liquid microextraction;HPLC-UV;鹽輔助均相液液微萃取;微生物發酵培養液樣品;HPLC-UV
出版社: 化學系所
引用: 1. Chen, H. L.; Chen, Y. S.; Juang, R. S., Separation of surfactin from fermentation broths by acid precipitation and two-stage dead-end ultrafiltration processes. Journal of membrane science 2007, 299 (1-2), 114-121. 2. Peypoux, F.; Bonmatin, J.; Wallach, J., Recent trends in the biochemistry of surfactin. Applied microbiology and biotechnology 1999, 51 (5), 553-563. 3. Banat, I. M.; Makkar, R. S.; Cameotra, S., Potential commercial applications of microbial surfactants. Applied microbiology and biotechnology 2000, 53 (5), 495-508. 4. Bognolo, G., Biosurfactants as emulsifying agents for hydrocarbons. Colloids and surfaces A: Physicochemical and engineering aspects 1999, 152 (1-2), 41-52. 5. Desai, J. D.; Banat, I. M., Microbial production of surfactants and their commercial potential. Microbiology and molecular Biology reviews 1997, 61 (1), 47-64. 6. Zhang, J.; Gorkovenko, A.; Gross, R. A.; Allen, A. L.; Kaplan, D., Incorporation of 2-hydroxyl fatty acids by< i> Acinetobacter calcoaceticus</i> RAG-1 to tailor emulsan structure. International journal of biological macromolecules 1997, 20 (1), 9-21. 7. Finnerty, W.; Singer, M., Microbial enhancement of oil recovery. Nature Biotechnology 1983, 1 (1), 47-54. 8. Makkar, R.; Cameotra, S., An update on the use of unconventional substrates for biosurfactant production and their new applications. Applied microbiology and biotechnology 2002, 58 (4), 428-434. 9. Jain, R. M.; Mody, K.; Mishra, A.; Jha, B., Isolation and structural characterization of biosurfactant produced by an alkaliphilic bacterium< i> Cronobacter sakazakii</i> isolated from oil contaminated wastewater. Carbohydrate Polymers 2011. 10. Arima, K.; Kakinuma, A.; Tamura, G., Surfactin, a crystalline peptidelipid surfactant produced by Bacillus subtilis: isolation, characterization and its inhibition of fibrin clot formation. Biochemical and biophysical research communications 1968, 31 (3), 488. 11. Shaligram, N. S.; Singhal, R. S., Surfactin!VA Review on Biosynthesis, Fermentation, Purification and Applications. Food Technology and Biotechnology 2010, 48 (2). 12. Chen, H. L.; Juang, R. S., Recovery and separation of surfactin from pretreated fermentation broths by physical and chemical extraction. Biochemical Engineering Journal 2008, 38 (1), 39-46. 13. Dufour, S.; Deleu, M.; Nott, K.; Wathelet, B.; Thonart, P.; Paquot, M., Hemolytic activity of new linear surfactin analogs in relation to their physico-chemical properties. Biochimica et Biophysica Acta (BBA)-General Subjects 2005, 1726 (1), 87-95. 14. Heerklotz, H.; Seelig, J., Detergent-like action of the antibiotic peptide surfactin on lipid membranes. Biophysical journal 2001, 81 (3), 1547-1554. 15. Chen, H. L.; Chen, Y. S.; Juang, R. S., Separation of surfactin from fermentation broths by acid precipitation and two-stage dead-end ultrafiltration processes. Journal of membrane science 2007, 299 (1), 114-121. 16. Shaligram, N. S.; Singhal, R. S., Surfactin–a review on biosynthesis, fermentation, purification and applications. Food Technol Biotechnol 2010, 48, 119-134. 17. Makkar, R.; Cameotra, S., Production of biosurfactant at mesophilic and thermophilic conditions by a strain of Bacillus subtilis. Journal of industrial microbiology & biotechnology 1998, 20 (1), 48-52. 18. Moran, A. C.; Martinez, M. A.; Sineriz, F., Quantification of surfactin in culture supernatants by hemolytic activity. Biotechnology letters 2002, 24 (3), 177-180. 19. Lin, S. C.; Jiang, H. J., Recovery and purification of the lipopeptide biosurfactant of Bacillus subtilis by ultrafiltration. Biotechnology techniques 1997, 11 (6), 413-416. 20. Banat, I. M., The isolation of a thermophilic biosurfactant producing Bacillus sp. Biotechnology letters 1993, 15 (6), 591-594. 21. Anthemidis, A. N.; Ioannou, K. I. G., Recent developments in homogeneous and dispersive liquid-liquid extraction for inorganic elements determination. A review. Talanta 2009, 80 (2), 413-421. 22. Anthemidis, A. N.; Ioannou, K. I. G., Recent developments in homogeneous and dispersive liquid–liquid extraction for inorganic elements determination. A review. Talanta 2009, 80 (2), 413-421. 23. Ghiasvand, A.; Shadabi, S.; Mohagheghzadeh, E.; Hashemi, P., Homogeneous liquid–liquid extraction method for the selective separation and preconcentration of ultra trace molybdenum. Talanta 2005, 66 (4), 912-916. 24. Ebrahimzadeh, H.; Yamini, Y.; Kamarei, F.; Shariati, S., Homogeneous liquid-liquid extraction of trace amounts of mononitrotoluenes from waste water samples. Analytica Chimica Acta 2007, 594 (1), 93-100. 25. Oence, D. N.; Gu, T., Liquid-liquid equilibrium of the acetonitrile-water system for protein purification. Separations Technology 1996, 6 (4), 261-264. 26. Murata, K.; Yokoyama, Y.; Ikeda, S., Homogeneous liquid-liquid extraction method. Extraction of iron (III) thenoyltrifluoroacetonate by propylene carbonate. Analytical Chemistry 1972, 44 (4), 805-810. 27. Matkovich, C. E.; Christian, G. D., Solvent extraction of metal chelates into water-immiscible acetone. Analytical Chemistry 1974, 46 (1), 102-106. 28. Tavakoli, L.; Yamini, Y.; Ebrahimzadeh, H.; Shariati, S., Homogeneous liquid-liquid extraction for preconcentration of polycyclic aromatic hydrocarbons using a water/methanol/chloroform ternary component system. Journal of Chromatography A 2008, 1196, 133-138. 29. Shamsipur, M.; Hassan, J., A novel miniaturized homogenous liquid-liquid solvent extraction-high performance liquid chromatographic-fluorescence method for determination of ultra traces of polycyclic aromatic hydrocarbons in sediment samples. Journal of Chromatography A 2010, 1217 (30), 4877-4882. 30. Rezaei, F.; Hosseini, M. R. M., New method based on combining ultrasonic assisted miniaturized matrix solid-phase dispersion and homogeneous liquid-liquid extraction for the determination of some organochlorinated pesticides in fish. Analytica Chimica Acta 2011. 31. Hassan, J.; Farahani, A.; Shamsipur, M., Rapid and simple low density miniaturized homogeneous liquid-liquid extraction and gas chromatography/mass spectrometry for determination of pesticide residues in sediment. Journal of Hazardous Materials 2010. 32. Da Silva, J. F.; Martins, W., Extraction of Fe (III), Cu (II), Co (II), Ni (II) and Pb (II) with thenoyltrifluoroacetone using the ternary solvent system water/ethanol/methylisobutylketone. Talanta 1992, 39 (10), 1307-1312. 33. Takagai, Y.; Kubota, T.; Akiyama, R.; Aoyama, E.; Igarashi, S., Preconcentration technique for nonylphenol using cellulose cotton with homogenous liquid–liquid extraction for liquid chromatographic analysis. Analytical and bioanalytical chemistry 2004, 380 (2), 351-354. 34. Carlos de Andrade, J.; Jose Cuelbas, C.; de Paula Eiras, S., Spectrophotometric determination of Mo (VI) in steel using a homogeneous ternary solvent system after single-phase extraction. Talanta 1998, 47 (3), 719-727. 35. Jamali, M. R.; Assadi, Y.; Shemirani, F., Homogeneous liquid–liquid extraction and determination of cobalt, copper, and nickel in water samples by flame atomic absorption spectrometry. Separation Science and Technology 2007, 42 (15), 3503-3515. 36. Igarashi, S.; Ide, N.; Takagai, Y., High-performance liquid chromatographic-spectrophotometric determination of copper (II) and palladium (II) with 5, 10, 15, 20-tetrakis (4N-pyridyl) porphine following homogeneous liquid-liquid extraction in the water-acetic acid-chloroform ternary solvent system. Analytica Chimica Acta 2000, 424 (2), 263-269. 37. Takahashi, A.; Ueki, Y.; Igarashi, S., Homogeneous liquid-liquid extraction of uranium (VI) from acetate aqueous solution. Analytica Chimica Acta 1999, 387 (1), 71-75. 38. Gupta, M.; Pillai, A. K. K. V.; Singh, A.; Jain, A.; Verma, K. K., Salt-assisted liquid-liquid microextraction for the determination of iodine in table salt by high-performance liquid chromatography-diode array detection. Food Chemistry 2010. 39. Shamsipur, M.; Hassan, J., A novel miniaturized homogenous liquid–liquid solvent extraction-high performance liquid chromatographic-fluorescence method for determination of ultra traces of polycyclic aromatic hydrocarbons in sediment samples. Journal of Chromatography A 2010, 1217 (30), 4877-4882. 40. Zhao, X.; Liu, X.; Zhao, Z.; Huang, C.; Zhang, M.; Wang, H.; Wang, X., Homogeneous liquid–liquid extraction combined with high performance liquid chromatography–fluorescence detection for determination of polycyclic aromatic hydrocarbons in vegetables. Journal of separation science 2009, 32 (12), 2051-2057. 41. Wang, X.; Zhao, X.; Liu, X.; Li, Y.; Fu, L.; Hu, J.; Huang, C., Homogeneous liquid–liquid extraction combined with gas chromatography–electron capture detector for the determination of three pesticide residues in soils. Analytica Chimica Acta 2008, 620 (1), 162-169. 42. Matkovich, C. E.; Christian, G. D., Salting-out of acetone from water. Basis of a new solvent extraction system. Analytical Chemistry 1973, 45 (11), 1915-1921. 43. Rice, N.; Irving, H.; Leonard, M., Nomenclature for liquid-liquid distribution (solvent extraction). Pure Appl. Chem 1993, 65 (11), 2373-2396. 44. Zhang, Y.; Cremer, P. S., Interactions between macromolecules and ions: the Hofmeister series. Current opinion in chemical biology 2006, 10 (6), 658-663. 45. Nucci, N. V.; Vanderkooi, J. M., Effects of salts of the Hofmeister series on the hydrogen bond network of water. Journal of molecular liquids 2008, 143 (2-3), 160-170. 46. Ni, N.; Yalkowsky, S. H., Prediction of Setschenow constants. International journal of pharmaceutics 2003, 254 (2), 167-172. 47. Xie, W. H.; Shiu, W. Y.; Mackay, D., A review of the effect of salts on the solubility of organic compounds in seawater. Marine Environmental Research 1997, 44 (4), 429-444. 48. Kunz, W.; Henle, J.; Ninham, B., ‘Zur Lehre von der Wirkung der Salze’(about the science of the effect of salts): Franz Hofmeister''s historical papers. Current opinion in colloid & interface science 2004, 9 (1), 19-37. 49. Menendez, J.; Arenillas, A.; Fidalgo, B.; Fernandez, Y.; Zubizarreta, L.; Calvo, E.; Bermudez, J., Microwave heating processes involving carbon materials. Fuel Processing Technology 2010, 91 (1), 1-8. 50. Iwaguch, S.; Matsumura, K.; Tokuoka, Y.; Wakui, S.; Kawashima, N., Sterilization system using microwave and UV light. Colloids and surfaces B: Biointerfaces 2002, 25 (4), 299-304. 51. Bogdal, D.; Prociak, A., Microwave-enhanced polymer chemistry and technology. Wiley: 2007. 52. Hayes, B. L., Microwave synthesis : chemistry at the speed of light. 2002; p p 295. 53. Maktabi, S.; Watson, I.; Parton, R., Synergistic effect of UV, laser and microwave radiation or conventional heating on E. coli and on some spoilage and pathogenic bacteria. Innovative Food Science & Emerging Technologies 2011. 54. Haque, K. E., Microwave energy for mineral treatment processes—a brief review. International Journal of Mineral Processing 1999, 57 (1), 1-24. 55. Yeh, M. S.; Wei, Y. H.; Chang, J. S., Bioreactor design for enhanced carrier-assisted surfactin production with Bacillus subtilis. Process Biochemistry 2006, 41 (8), 1799-1805. 56. Doong, R.; Chang, S.; Sun, Y., Solid-phase microextraction for determining the distribution of sixteen US Environmental Protection Agency polycyclic aromatic hydrocarbons in water samples. Journal of Chromatography A 2000, 879 (2), 177-188. 57. 莊壬豪. 微波輔助衍生化結合頂空固相微萃取技術以GC-ECD偵測生質柴油中的微量甘油. 2009. 58. Su, Y. S.; Jen, J., Determination of organophosphorous pesticides in water using in-syringe ultrasound-assisted emulsification and gas chromatography with electron-capture detection. Journal of Chromatography A 2010, 1217 (31), 5043-5049. 59. Tsai, K. P.; Chen, C. Y., An algal toxicity database of organic toxicants derived by a closed‐system technique. Environmental Toxicology and Chemistry 2007, 26 (9), 1931-1939. 60. Goss, F. R., 173. The magnitude of the solvent effect in dipole-moment measurements. Part III. Polarisation and association of alcohols in the liquid phase. J. Chem. Soc. 1940, (0), 888-894. 61. Apelblat, A., The vapour pressures of saturated aqueous solutions of potassium bromide, ammonium sulfate, copper (II) sulfate, iron (II) sulfate, and manganese (II) dichloride, at temperatures from 283 K to 308 K. The Journal of Chemical Thermodynamics 1993, 25 (12), 1513-1520. 62. Tang, J. S.; Zhao, F.; Gao, H.; Dai, Y.; Yao, Z. H.; Hong, K.; Li, J.; Ye, W. C.; Yao, X. S., Characterization and Online Detection of Surfactin Isomers Based on HPLC-MSn Analyses and Their Inhibitory Effects on the Overproduction of Nitric Oxide and the Release of TNF-α and IL-6 in LPS-Induced Macrophages. Marine drugs 2010, 8 (10), 2605-2618. 63. PEYPOUX, F.; BONMATIN, J. M.; LABBE, H.; DAS, B. C.; PTAK, M.; MICHEL, G., Isolation and characterization of a new variant of surfactin, the [Val7] surfactin. European Journal of Biochemistry 1991, 202 (1), 101-106. 64. Wei, Y. H.; Chu, I., Enhancement of surfactin production in iron-enriched media by Bacillus subtilis ATCC 21332. Enzyme and microbial technology 1998, 22 (8), 724-728.
本研究開發出以鹽輔助均相液液微萃取之樣品前處理技術,並以HPLC-UV分離偵測bacillius subtilis CWS1微生物發酵培養液中的表面素(surfactin)。表面素為一種生物界面活性劑,具有高表面活性、易被生物降解暨對環境友善等優點,因此在未來有極大潛力取代合成的化學界面活性劑成為產品化商品。在產品量化生產過程中,產品的穩定性影響其有效性,為了能監測微生物培養液中表面素的濃度,快速地分析表面素技術是十分重要的。本研究使用實驗室自行設計的玻璃萃取裝置,將與水互溶的有機溶劑及稀釋過的培養液放入萃取裝置之後,加入鹽類並攪拌使溶液達飽和,之後將裝置至於離心機離心,最後取上層液打入HPLC進行分析。在鹽輔助均相液液微萃取技術中,針對萃取溶劑、萃取溶劑體積、添加的鹽類、硫酸銨鹽添加量、及水樣的pH值等參數進行探討。實驗結果顯示,5毫升經pH2的緩衝溶液稀釋的培養液加入300μL的Acetonitrile及4克硫酸銨鹽,攪拌之後離心1分鐘會有最好的萃取效果。本方法測得培養液中表面素的線性範圍為2-250 mg/L ,線性相關係數介於0.9970-0.9994之間,應用培養液在真實樣品中的回收率為77.7%~106.7%。此方法對於偵測微生物發酵培養液中的表面素是一個簡單、快速、省錢、對環境友善、及少基質干擾的分析方法。

In this study, a simple, rapid and efficient sample pretreatment method has been developed, termed new in-tube salt-assisted homogeneous liquid-liquid microextraction (SHLLME) followed by high performance liquid chromatography/ultra-violet detection (HPLC/UV) for the determination of surfactin in microbial fermentation broths of bacterium bacillius subtilis CWS1. Surfactin is one of a biosurfactant and it has the potential to replace synthetic chemical surfactant in the near future because of its high surface activity, biodegradability and environmental-friendly nature. In order to characterize the amount of surfactin for the microbial monitoring of fermentation production process, we have demonstrated a new method to analyze surfactin using a home-made extraction device. In this method, an extractant (water-miscible organic solvents) and salt were added to broth sample in an extraction tube. Then the device was sealed with plunger cork and taken for stirring and centrifugation processes. After that, the amount of separated extractant was measured using self-scaled glass capillary tube which was in-printed in the extraction device. Then the collected extractant was injected directly into HPLC-UV for analysis. Parameters influencing the extraction efficiency of SHLLE-HPLC/UV, such as the extraction solvent, solvent volume, pH, and salt-addition were thoroughly investigated and optimized. Under the selected experimental conditions, surfactin was extracted from 5 mL broth sample (at pH 2) by the addition of 300 μL acetonitrile (extractant) and 4 g ammonium sulfate, and followed by stirring for 5 min, and then centrifugation for 1 min at 3200 rpm, yields the maximum extraction efficiency. The proposed method provides a simple, rapid, inexpensive, eco-friendly and less matrix interferences process for the determination surfactin in microbial fermentation broth samples.
其他識別: U0005-0607201217253500
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