一种快速提取分析微囊藻毒素的方法
A RAPID EXTRACTING AND ANALYZING METHOD FOR MICROCYSTIN
-
摘要: 以五种群体和单细胞微囊藻及自然微囊藻水华为材料,在沸水浴中经不同时间处理,过滤后直接进行HPLC-UV检测,发现12min的沸水浴处理就足以达到抽提目的.研究中发现,去离子水比蒸馏水是更有效的抽提溶剂.传统的甲醇抽提结果与沸水浴处理的相对误差主要在0.2%-16.59%之间.结果还显示,群体微囊藻需要比单细胞微囊藻抽提更长时间.本研究提供了一种经过改进的高效、廉价和快速的微囊藻毒素抽提分析方法.Abstract: Microcystins are a group of natural toxins known to be produced by certain types of freshwater blue-green algae and which have been shown to be liver toxins and tumour promoters, and pose a health hazard for humans, domestic animals and wildlife. The traditional methods for toxin extraction include 5% acetic acid, 75% methanol, 100% methanol, 70% acetonitrile and so on. The extracting solution using traditional methods should be firstly pass through a sep-pak cartridge, then was rinsed with water and 25% methanol in water, and then microcystins were eluted with 90% methanol water, which is very overelaborate and expensive. In this paper, a new rapid method for toxin extraction was provided based on boiling-water bath, and this method were used to extract cultures of five strains of colonial or single-celled Microcystis and one natural water bloom sample. With different times dealing in boiling-water bath, and through 0.45μm membrane, it was detected directly by HPLC-UV, and it was found that twelve minutes boiling-water bath was enough for microcystin extraction. Comparing with the traditional extraction methods, the comparative error of boiling-water bath extraction method was from 0.2% to 16.59%. The results illustrated that more time was needed for extracting toxins from colonial Microcystis than from single-celled one. This study provides an efficient, cheap and quick extracting method for microcystin, which reveals a comprehensive and applied prospect.
-
Keywords:
- Microcystis /
- Microcystin /
- Boiling-water bath /
- HPLC
-
-
[1] He Z R. Seven microcystins from Microcystis water bloom in Lake Dalai, [J]. China. J. Envir. Sci., 1997,9(1):113-119[2] Wu Weinang The study on the major algal in lake Dianchi[J]. Yunnan Environmental Science, 1997, 16(2):26-29. [吴为梁. 滇池水体中主要藻种毒素研究[J]. 云南环境科学,1997,16(2):26-29][3] Tang Z. Y., Wu M. C Xia S. S. Prmary liver cancer [M], Beijing:China Academic Publishers and Springer-Verlag, 1989,30-37[4] Codd G A. Cyanobacterial toxins:oceurrcnce, properties and significance [J]. Water Sci. Technol., 1995,32:149-156[5] Ahluwalia K B Culture of the organism that causes rhinosporidiosis[J]. J. Laryngol. Otol., 1999,113(6):523-528[6] Jussi M Chromatography of microcystins [J]. Analytica. Chimica. Acta., 1997,352:277-298[7] Metcalf J S, Codd G Microwave oven and boiling waterbath extraction of hepatotoxins from cyanobacterial cells[J]. FEMS Microbiol. Lett., 2000,184:241-246[8] Nishizawa K, Chihara M. Methods in Algological Studies. Kyoritsu[M]. Tokyo,1979,294-305[9] Song L R, Lei L M, He Z R, et al. The growth and physiologic characteristics of blue Microcystis. Aeruginosa and green microcystis and the analysis of microcystins from water lboom in lake Dianchi. Acta Hydrobiologica Sinica, 1999, 23(3):402-408. [宋立荣,雷腊梅,何振荣,等. 滇池水华蓝藻铜锈微囊藻和绿色微囊藻的生长生理特性及毒素分析[J]. 水生生物学报,1999,23(3):402-408][10] Honkanan R E, Codispoti B A, Tse K, et al. Characterization of natural toxins with inhibitory activity against serine/threonine protein phosphatases[J]. Toxicon,1994,32(3):339-350[11] Ward C J, Beattie K A, Lee E Y C, et al. Colorimetric protein phosphatase inhibition assay of laboratory strains and natural blooms of cyanobacteria: comparisons with high-performance liquid chromatographic analysis for microcystins[J]. FEMS Microbiol. Lett. 1997, 153:465-473[12] Chu F S, Huang X, Wei R D, et al Production and characterization of antibodies against microcystins[J]. Appl. Envir. Microbiol., 1989,55:1928-1933[13] Chu F S, Huang X, Wei R D. Enzyme-linked immunosorbent assay for microcystins in blue-green algal blooms[J]. J. Assoc. Off. Analyt. Chem.,1990,73:451-456[14] Onyewuenyi N, Hawkins P. Separation of toxic peptide(microcystins) in capillary electrophoresis, with the aid of organic mobile phase modifiers[J]. J. Chromatogr.,1995,749:271-277[15] Hui Pan, Lirong Song, Yonding Liu and Thomas Borner. Detection of Hepatotoxic Microcystis Strains by PCR with Intact Cells from both Culture and Environmental Samples [J]. Archiv Microbiology, 2002, 178:421-427 He Z R. Seven microcystins from Microcystis water bloom in Lake Dalai, [J]. China. J. Envir. Sci., 1997,9(1):113-119[2] Wu Weinang The study on the major algal in lake Dianchi[J]. Yunnan Environmental Science, 1997, 16(2):26-29. [吴为梁. 滇池水体中主要藻种毒素研究[J]. 云南环境科学,1997,16(2):26-29][3] Tang Z. Y., Wu M. C Xia S. S. Prmary liver cancer [M], Beijing:China Academic Publishers and Springer-Verlag, 1989,30-37[4] Codd G A. Cyanobacterial toxins:oceurrcnce, properties and significance [J]. Water Sci. Technol., 1995,32:149-156[5] Ahluwalia K B Culture of the organism that causes rhinosporidiosis[J]. J. Laryngol. Otol., 1999,113(6):523-528[6] Jussi M Chromatography of microcystins [J]. Analytica. Chimica. Acta., 1997,352:277-298[7] Metcalf J S, Codd G Microwave oven and boiling waterbath extraction of hepatotoxins from cyanobacterial cells[J]. FEMS Microbiol. Lett., 2000,184:241-246[8] Nishizawa K, Chihara M. Methods in Algological Studies. Kyoritsu[M]. Tokyo,1979,294-305[9] Song L R, Lei L M, He Z R, et al. The growth and physiologic characteristics of blue Microcystis. Aeruginosa and green microcystis and the analysis of microcystins from water lboom in lake Dianchi. Acta Hydrobiologica Sinica, 1999, 23(3):402-408. [宋立荣,雷腊梅,何振荣,等. 滇池水华蓝藻铜锈微囊藻和绿色微囊藻的生长生理特性及毒素分析[J]. 水生生物学报,1999,23(3):402-408][10] Honkanan R E, Codispoti B A, Tse K, et al. Characterization of natural toxins with inhibitory activity against serine/threonine protein phosphatases[J]. Toxicon,1994,32(3):339-350[11] Ward C J, Beattie K A, Lee E Y C, et al. Colorimetric protein phosphatase inhibition assay of laboratory strains and natural blooms of cyanobacteria: comparisons with high-performance liquid chromatographic analysis for microcystins[J]. FEMS Microbiol. Lett. 1997, 153:465-473[12] Chu F S, Huang X, Wei R D, et al Production and characterization of antibodies against microcystins[J]. Appl. Envir. Microbiol., 1989,55:1928-1933[13] Chu F S, Huang X, Wei R D. Enzyme-linked immunosorbent assay for microcystins in blue-green algal blooms[J]. J. Assoc. Off. Analyt. Chem.,1990,73:451-456[14] Onyewuenyi N, Hawkins P. Separation of toxic peptide(microcystins) in capillary electrophoresis, with the aid of organic mobile phase modifiers[J]. J. Chromatogr.,1995,749:271-277[15] Hui Pan, Lirong Song, Yonding Liu and Thomas Borner. Detection of Hepatotoxic Microcystis Strains by PCR with Intact Cells from both Culture and Environmental Samples [J]. Archiv Microbiology, 2002, 178:421-427
计量
- 文章访问数: 828
- HTML全文浏览量: 0
- PDF下载量: 444
下载: