Cfrench:anthrone

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(Assaying polysaccharides with the anthrone assay)
(Assaying polysaccharides with the anthrone assay)
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'''Equipment and Instrument'''
'''Equipment and Instrument'''
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*1. 19×150mm glass test tube
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1. 19×150mm glass test tube
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*2. Cuvettes
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2. Cuvettes
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*3. Glass pipette and pipette aid
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3. Glass pipette and pipette aid
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*4. Spectrometer
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4. Spectrometer
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*5. Chemical hood
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5. Chemical hood
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'''Preparation of Reagent and Standard Solution'''
'''Preparation of Reagent and Standard Solution'''
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*1. 2% anthrone in ethyl acetate ['''SAFETY NOTES''': Ethyl acetate is highly flammable in presence of open flames and sparks, of heat. Be careful when using it.]
+
1. 2% anthrone in ethyl acetate ['''SAFETY NOTES''': Ethyl acetate is highly flammable in presence of open flames and sparks, of heat. Be careful when using it.]
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*2. Glucose standard solution (0~80µg):
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2. Glucose standard solution (0~80µg):
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**SolA: 1mg/2mL std. (100mL): 50mg glucose + 100mL H2O (50mg glucose in total)
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SolA: 1mg/2mL std. (100mL): 50mg glucose + 100mL H2O (50mg glucose in total)
    
    
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**SolB: 100µg/2mL std. (50mL): 5mL SolA + 45mL H2O (2.5mg glucose in total)
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SolB: 100µg/2mL std. (50mL): 5mL SolA + 45mL H2O (2.5mg glucose in total)
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  1) 80µg/2mL std. (10mL): 8mL SolB + 2mL H2O (400µg glucose in total)
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*1) 80µg/2mL std. (10mL): 8mL SolB + 2mL H2O (400µg glucose in total)
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  2) 50µg/2mL std. (20mL): 10mL SolB + 10mL H2O (500µg glucose in total)
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*2) 50µg/2mL std. (20mL): 10mL SolB + 10mL H2O (500µg glucose in total)
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  3) 25µg/2mL std. (20mL): 5mL SolB + 15mL H2O (250µg glucose in total)
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*3) 25µg/2mL std. (20mL): 5mL SolB + 15mL H2O (250µg glucose in total)
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  4) 10µg/2mL std. (20mL): 2mL SolB + 18mL H2O (100µg glucose in total)
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*4) 10µg/2mL std. (20mL): 2mL SolB + 18mL H2O (100µg glucose in total)
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  5) 5µg/2mL std, (20mL): 1mL SolB + 19mL H2O (50µg glucose in total)
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*5) 5µg/2mL std, (20mL): 1mL SolB + 19mL H2O (50µg glucose in total)
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  (All the standard solutions can store in refrigerator or freezer to re-use.)
+
(All the standard solutions can store in refrigerator or freezer to re-use.)

Revision as of 18:37, 23 August 2008

Assaying polysaccharides with the anthrone assay

Equipment and Instrument

1. 19×150mm glass test tube

2. Cuvettes

3. Glass pipette and pipette aid

4. Spectrometer

5. Chemical hood


Preparation of Reagent and Standard Solution

1. 2% anthrone in ethyl acetate [SAFETY NOTES: Ethyl acetate is highly flammable in presence of open flames and sparks, of heat. Be careful when using it.]

2. Glucose standard solution (0~80µg):

SolA: 1mg/2mL std. (100mL): 50mg glucose + 100mL H2O (50mg glucose in total)

SolB: 100µg/2mL std. (50mL): 5mL SolA + 45mL H2O (2.5mg glucose in total)

  • 1) 80µg/2mL std. (10mL): 8mL SolB + 2mL H2O (400µg glucose in total)
  • 2) 50µg/2mL std. (20mL): 10mL SolB + 10mL H2O (500µg glucose in total)
  • 3) 25µg/2mL std. (20mL): 5mL SolB + 15mL H2O (250µg glucose in total)
  • 4) 10µg/2mL std. (20mL): 2mL SolB + 18mL H2O (100µg glucose in total)
  • 5) 5µg/2mL std, (20mL): 1mL SolB + 19mL H2O (50µg glucose in total)

(All the standard solutions can store in refrigerator or freezer to re-use.)


Procedure

1.Add 2mL samples or standards [0(pure water), 5, 10, 25, 50, 80µg/2mL] solution into all prepared test tubes.

2.(In chemical hood) Use glass pipette and gently add 0.5mL 2% anthrone solution into each test tube. There will be two layers of this solution. The upper layer is anthrone and the lower layer is the sample/standard. Try not to disturb solution, because ethyl acetate is easy to vaporize.

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