JAK PRZETESTOWAĆ POLISACHARYDÓW

Jak przetestować pneumokokom?

1.试剂Reagent

1.1苯酚 fenolu (AR)

1.2重蒸水RedistilledWater

1.3浓硫酸 SulfuricAcid (AR)

1.45%苯酚溶液:取苯酚加热蒸馏,收集182度馏分,吸取此馏分10ml加水至200ml即可。

5% PhenolSolution: Takeheatphenoldistillationanddistillatecollect182degree, absorbdistillation10mlandaddwaterto200ml.

2.仪器和用具InstrumentandAppliance

2.1电子天平 ElectronicBalance (1/100000)

2.2玻璃仪器:容量瓶GlassApparatus:VolumetricFlask

2.3超声波清洗器UltrasonicCleaner

2.4紫外分光光度计UltravioletSpectrophotometer

2.5玻璃比色皿 GlassCell (25 pikseli)

3、步骤Process

3.1标准曲线的建立

EstablishedStandardCurves

3.1.1精密称取恒重后的葡萄糖对照品10mg置于100ml容量瓶中,加入重蒸水溶解并定容。

AccuratelymeasureconstantweightGlucosereferencesubstance10mgandadd into100mlvolumetricflaskwithheavysteamwaterdissolvedandconstantvolume.

3.1.2用吸管精密吸取上述标准液0,0.1,0.2,0.4,0.6,0.8ml。 分别置于具塞刻度试管中,并用蒸馏水补充至2ml,加入5%的苯酚溶液1.0ml,浓硫酸5.0ml,振摇5分钟,同时作一个空白对照。

Accuratelyabsorbabovestandardliquid0, 0,1, 0,2, 0,4, 0,6, 0,8 ml, respectivelyin Graduatedtesttubewithstopper,adddistilledwaterto2ml,andadd1.0ml5% PhenolSolution, SulfuricAcid5.0ml, Vibrationwave5minutes, makeablank controlatthesametime.

3.1.3在沸水浴中加热15分钟,取出,冷水冷却30分钟。

Heatingfor15minutesinaboilingwater, takeoutandcoolingincoldwaterfor30 minut.

3.1.4紫外可见分光光度计在波长490nm处测定吸光度。

UV-visspectrophotometerabsorbanceat490nmwavelength.

3.1.5以吸光度为纵坐标,葡萄糖浓度为横坐标绘制标准曲线。

Absorbanceasordinate, glucoseconcentrationasabscissa, drawingstandardcurve.

3.2样品的测定SampleTest

3.2.1 精密称取样品0.20g,置索氏提取器中,用80%乙醇10ml回流2h,残渣用

80% 乙醇洗至无色,, 继续用水100ml回流2h, 反复洗至250ml容量瓶中, 定容, 摇匀成为样品液。

Accuratelymeasuresample0.2gintosoxhletextractor,with10ml80%ethanol reflux2h, andResiduewith80% ethanoltowashtocolorless, thenwith100ml waterreflux2h,

repeatedwashinginto250mlvolumetricflask, constantvolume, Shakewelltothesampleliquid.

3.2.2 精密吸取上述溶液2.0ml置具塞试管中,照标准曲线项下"加入5%的苯酚溶液~"起,依法操作

Accuratelyabsorbabovestandardliquid 2.0 ml, addintoGraduatedtesttubewith korek, Accordingtothestandardcurveandquot; dodać 5% PhenolSolutionandquot;

3.2.3 在标准曲线上查出样品吸光度值对应的毫克数。

Findoutthesamplenumberofmgabsorbancevalues

4、计算Calculate

M--SampleVolume(g);

V--Dilutionratio;

A--Sampleabsorbancecorrespondingmg