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wdmzswpy

新虫 (小有名气)

[求助] 关于胞外聚合物(EPS)的一些问题~ 已有1人参与

想从活性污泥里面提取EP,现在遇到几个问题,希望能得到大神和好心人的解答~
       看了文献用阳离子交换树脂来提,想提取SEPS,LB-EPS和TB-EPS 这三种成分。方法上是先用了20ml的泥样(就是SBR反应器搅拌以后直接抽出来的20ml,MLVSS在2300左右)加去离子水至45ml,然后经过超声、离心、抽滤得到SEPS。
       老师说EPS溶液和酒精1:4混合以后,如果有EPS会形成絮状沉淀,但是我试过基本上看不出来絮状沉淀,有也只是很小很小一点,不知道是不是其他杂质飘到里面。想问下大家这样可以判断溶液里面有无EPS吗,还有其他方法可以检测出来溶液里面有没有EPS存在吗?
       还有就是我想把EPS从溶液中提取出来,现在是用冷冻干燥,这样试过结果也基本看不见EPS,是我的污泥量不够吗?
PS:以下是我参考的提取方法:
        Fresh biofilm was scraped from carriers with a blade, and S-sludge was collected from suspension of the reactor. The SEPS and LB-EPS of the biofilm and S-sludge samples were extracted using the modified method described by Liang et al. (2010). Approximately 20 mL of fresh samples were placed in 50 mL centrifuge tubes, mixed with deionized water to form a 45 mL suspension, and then treated by ultrasound at 20 kHz and 40 W for 30 s. The suspensions were then centrifuged at 2000g for 15 min at 4 °C. SEPS were collected from the supernatant. The sediments at the bottom of the centrifuge tubes were resuspended to their initial volume with deionized water. The suspensions were horizontally vibrated in a thermostat incubator for 1 h and then centrifuged at 5000g for 15 min at 4 °C. LB-EPS were collected from the supernatant. After the SEPS and LB-EPS were extracted, the residual sediments were resuspended to their initial volume again with phosphate buffer saline (pH 7.5) to extract TB-EPS. The TB-EPS were extracted by cationic exchange resin (CER). 70 g g−1 volatile suspended solids of gel CER was added to the suspension and stirred at 600 rpm for 1 h at 4 °C. The suspensions were centrifuged at 10 000g for 15 min at 4 °C. All the supernatant were filtered through 0.45 μm filters and stored at −20 °C before analysis. The duplicate extraction tests were conducted in this work.
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yalisi1976

木虫 (小有名气)

【答案】应助回帖


wdmzswpy(gaofeng925代发): 金币+1, 积极交流 2016-12-03 22:38:14
如果冷冻干燥后还看不到里面有EPS固体的话,说明EPS浓度太低了。溶解型EPS的含量变化波动比较大,如果你用CER等方法提取了结合型EPS的话,肯定能提取出来的。
2楼2016-12-03 13:55:38
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wdmzswpy

新虫 (小有名气)

引用回帖:
2楼: Originally posted by yalisi1976 at 2016-12-03 13:55:38
如果冷冻干燥后还看不到里面有EPS固体的话,说明EPS浓度太低了。溶解型EPS的含量变化波动比较大,如果你用CER等方法提取了结合型EPS的话,肯定能提取出来的。

谢谢亲~  今天刚提了LB-eps,但是我按照1:4加酒精还是看不见絮体,这种方法能判断溶液里有EPS吗? 另外由于实验室离心机不能控制温度,没有按照方法要求在4℃离心,这个对提取影响大吗?
3楼2016-12-04 17:17:01
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yalisi1976

木虫 (小有名气)

引用回帖:
3楼: Originally posted by wdmzswpy at 2016-12-04 17:17:01
谢谢亲~  今天刚提了LB-eps,但是我按照1:4加酒精还是看不见絮体,这种方法能判断溶液里有EPS吗? 另外由于实验室离心机不能控制温度,没有按照方法要求在4℃离心,这个对提取影响大吗?...

浓度过低的话,加75%的酒精比例仍然看不到沉淀的。文献中这种方法提到的LB是很低的。如果你一定要看到沉淀物,还得提TB
4楼2016-12-04 20:43:34
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