纤维素提取方法教案资料课件.ppt

1、纤维素提取方法简介How could we extract cellulose from tree ring-step by stepPeili FU(付培立)March 14th 2014references Wieloch T,Helle G,Heinrich I,et al.A novel device for batch-wise isolation of-cellulose from small-amount wholewood samplesJ.Dendrochronologia,2011,29(2):115-117.Laumer W,Andreu L,Helle G,et al.

2、A novel approach for the homogenization of cellulose to use micro amounts for stable isotope analysesJ.Rapid Communications in Mass Spectrometry,2009,23(13):1934-1940.样品的切取和称量 Cutting and weighing样品大小：厚度 thickness 0.5-0.8 mm;sample size 500m样品量 sample amount：2 mg Samples needed for mass spectrometry

3、:one measurement of:C13 0.5mg；O18 0.3 mg；(production 30%)提取 extraction NaOH 5%solution 溶液,60,2h,2 times次;（近沸腾热的去离子水，洗3次,washed 3 times with boiled de-ioned water）NaClO2 7%solution 溶液,60,8h,6 times（醋酸溶液 acetic acid,调节pH至 4-5）;（近沸腾热的去离子水，洗3次,washed 3 times with de-ioned boiled water）NaOH 17%solution 溶

4、液,室温室温(room temperature),2h,2 times次 （1%HCl 溶液洗1次，近沸腾热的去离子水，洗3次 测pH,washed with 1%HCl,then washed 3 times with de-ioned boiled water）判断是否提取完全(completely extracted?)草酸溶液:每100ml NaClO2 加草酸溶液 0.51ml草酸 acetic acid solutions:0.5ml1ml/100ml NaClO2 7%纤维素转移 cellulose transferEppi管清洗(高压气体、去离子水）、加入1ml去离子水；纤维素

5、转移镊子 去离子水等冰箱4 冷藏纤维素的同质化 homogenization 样品冷藏 store in refrigerator(+4）方法：超声波；ultrasonic 设置：cycle 0.5，amplitude 6-7；继续时间 duration 30s（13C 2*15s 18O 30s 因不同物种而有所差异）；判断 呈牛奶状(milky)，或者纤维素均一(or homogenized盖上带孔的盖子，至于冰箱中冷冻(为低温真空干燥作准备)stored in refrigerator-10真空低温干燥(freeze dry)关闭排气阀、排水阀，检查油泵中油量多少。打开机器和油泵预热30

6、分钟（warm up）；将样品放入，一般一次可放入3个eppi板（288个样品左右），主干燥程序，48h；真空度 atm 降至 1.030mbar(main drying process)调节程序至干燥后程序，24h；(after dry process)真空 1.030mbar 降至 0.0030.005mbar 调节至待机程序 小心打开排气阀，使压力升至1atm 然后取出样品，打开排水阀，关闭机器和泵纤维素称量 weigh of cellulose 使用半微量天平（与称量木材使用同一天平）精度 0.01mg同位素测定 premeasurement 需要量 the amount of cel

7、lulose needed:18O 300g，13C 500g 银（18O,Silver capsule）或者钛（13C,Titanium capsule）胶囊裹住；再次干燥：高温、真空将所有的水除去 remove all the water in samples;测定：质谱仪 spectrometryDry befroe analysis with mass spectrometrySamples for mass spectrometry:O18 silver capsule;C13 Titanium capsule废液处理 The waste NaClO2 7%注意溶液有气体产生,废液应敞口至于通风橱一段时间,然后再拧紧 Gas will emit from the wasted solution of NaClO2 7%,keep the lipid open for 35 days NaOH 17%容器的清洗 clean of filters500 OvenClean machineChrismas party