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Table 1 Summary of the changes from the original MBTH1 method to be adapted to the HT2 format

From: Automated saccharification assay for determination of digestibility in plant materials

Reagent Original method Modification for HT assay
  Volume of reagent 3 Concentration 3 Volume of reagent 4 Concentration 3
Sample 100 μl - 75 μl -
NaOH 100 μl 0.5 N 25 μl 1 N
MBTH mixture 100 μl 1.5 mg/ml MBTH
0.5 mg/ml
DTT
50 μl 0.21 mg/ml MBTH
0.7 mg/ml
DTT
Oxidising Reagent 200 μl 0.5% (FeNH4(SO4)2)
0.5% Sulfamic acid
0.25 N HCl
100 μl 0.5% (FeNH4(SO4)2)
0.5% Sulfamic acid
0.25 N HCl
H2O 500 μl - - -
Heating 80°C for 15 min - 60°C for 20 min -
Range of detection 0 to 20 nmol 0 to 200 nmol
  1. 1HT = high throughput.
  2. 2MBTH = 3-methyl-2benzothiazolinonehydrazone.
  3. 3Anton and Barrett [20].
  4. 4HT determination.