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7. Reducing sugar determination by Park and Johnson Method<br />

7.1 Apparatus and reagent<br />

204<br />

7.1.1 Spectrophotometer<br />

7.1.2 Boiling bath and ice bath<br />

7.1.3 Balance with digital<br />

7.1.4 Vortex mixer<br />

7.1.5 Cuvettes tubes for spectrophotometer<br />

7.1.6 Glassware (Erlenmeryer flask, volumetric flasks, pipette, test tube)<br />

7.1.7 Mechanical, adjustable volume pipettor (500 and 100 µl) with tips<br />

7.1.8 D-Glucose (C6 H12 O6) for standard glucose solution, 0-5 µg/ml.<br />

Prepare by adding 0.02 gram of dried D-glucose in distilled water<br />

and adjusted volume to 100 ml in volumetric flask.<br />

7.1.9 NaCO3 –NaHCO3-KCN reagent (for 1 lit preparation)<br />

- Potassium cyanide (KCN) 0.65 gram<br />

- Sodium carbonate (Na2 CO3 ) 4.80 gram<br />

- Sodium hydrogen carbonate (HNaO3) 9.20 gram<br />

Prepare by dissolve the Na2CO3 KCN chemical reagent into<br />

distilled water and adjusted volume to 1000 ml in volumetric flask.<br />

7.1.10 Ferriccyanide reagent (for 1 lit preparation)<br />

- Potassium ferriccyanide (K3Fe (CN))6 0.50 garm<br />

Prepare by dissolve the Ferriccyanide reagent into<br />

distilled water and adjusted volume to 1000 ml in volumetric flask.<br />

7.1.11 Ferric ammonium sulphate reagent (for 1 lit preparation)<br />

- FeNH4 (SO4)2.12H2O 3.0 gram<br />

- Sulphuric acid (H2SO4) 50 mM 1000 ml<br />

Prepare by dissolve FeNH4 (SO4)2.12H2O into Sulphuric acid<br />

50 mM and adjusted volume to 1000 ml in volumetric flask with<br />

50 mM Sulphuric acid.

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