Yin Li-hong, Zhang Guo-zhou, Cheng Guo-guang, Wei Si-shun
Key Laboratory of the Public Health and Safety, Ministry of Education, Department of Environmental Health, School of Public Health, Fudan University, Shanghai 200032, China.
Wei Sheng Yan Jiu. 2006 Mar;35(2):162-4.
To optimize the solid-phase extraction method of isolation, purification and determination of intracellular microcystin-RR, -YR and -LR (MC-RR,-YR,-LR), algal samples were collected from Dianshan Lake, which is a major water source of Shanghai.
During extraction, Algal samples were treated with different solvent (5% acetic acid and 75% methanol), at different temperature (room temperature and 56 degrees C), centrifuge at 10000 x g, 30 min and 3500 rpm/min, 10 min, eluted with 80% methanol with or without trifluoroacetate acid(TFA). Further, gradient elution were employed to separate microcystins in high performance liquid chromatography (HPLC), the mobile phase consisted of aqueous acetonitrile and 0.05% TFA.
When dried algae cells were extracted by 5% acetic acid, 56 degrees C water bath combined with centrifuge at 3500 rpm/min was an effective and simple method for the extraction of microcystin-RR, elutant contains TFA can markedly increase the yield of microcystins extraction; the concentration of microcystin-RR in nature algal samples was higher than MC-LR and MC-YR.
The extraction efficacy of MC-YR and -LR is higher in 75% methanol aqueous and eluted with 80% methanol containing 0.05% TFA, while the extraction efficacy of MC-RR is higher in 5% acetic acid.
为优化细胞内微囊藻毒素-RR、-YR和-LR（MC-RR、-YR、-LR）的分离、纯化及测定的固相萃取方法，采集了上海主要水源地淀山湖的藻类样本。
萃取过程中，藻类样本用不同溶剂（5%乙酸和75%甲醇）、在不同温度（室温及56℃）下处理，分别以10000×g离心30分钟和3500转/分钟离心10分钟，用含或不含三氟乙酸（TFA）的80%甲醇洗脱。此外，在高效液相色谱（HPLC）中采用梯度洗脱分离微囊藻毒素，流动相由乙腈水溶液和0.05% TFA组成。
当干燥藻细胞用5%乙酸萃取时，56℃水浴结合3500转/分钟离心是萃取微囊藻毒素-RR的有效且简便的方法，含TFA的洗脱剂可显著提高微囊藻毒素的萃取产量；天然藻类样本中微囊藻毒素-RR的浓度高于MC-LR和MC-YR。
MC-YR和-LR在75%甲醇水溶液中并用含0.05% TFA的80%甲醇洗脱时萃取效率较高，而MC-RR在5%乙酸中萃取效率较高。
