How long does it take to perform the Total Dietary Fiber Assay (K-TDFR)?

This is a guide to the analysis time and schedule for the procedure of the Total Dietary Fiber Assay Kit (K-TDFR):

Sample Preparation:

• Preparation of samples (possible for 6 samples in duplicate + duplicate blank) (1 g sample + 40 mL Buffer) 30 minutes
• Alpha-amylase incubation at 98-100 degrees. 30 minutes
• Cool to 60 degrees. 15 minutes
• Protease incubation. 30 minutes
• pH check/adjustment. 15 minutes
• Amyloglucosidase incubation. 30 minutes
• Weigh Celite into crucibles, dry for 1 hour, allow to cool. 2 hours
• Prepare crucibles for filtering. 30 minutes

Total time = 5 hours

Insoluble Dietary Fiber Analysis:

• Filter samples. (can filter 7 samples at once using manifold) Filtering depends on sample. Allow for 2-3 hours.
• Dry crucibles overnight
• Weigh cooled crucibles. 30 minutes.
• Protein determination. 4 hours
• Ash determination. Incinerate sample overnight.
• Weigh cooled ashed sample. 30 minutes

Soluble Dietary Fiber:

• Same sample preparation – Total of 5 hours.
• Filter samples as per IDF. Allow 2-3 hours.
• Combine filtrate and washings. Add 95% EtOH. Allow to precipitate. 60 minutes.
• Weigh Celite into crucibles, dry for 1 hour, allow to cool. 2 hours
• Prepare crucibles for filtering. 30 minutes
• Filter samples. (can filter 7 samples at once using manifold) Filtering depends on sample. Allow for 2-3 hours.
• Dry crucibles overnight
• Weigh cooled crucibles. 30 minutes.
• Protein determination. 4 hours
• Ash determination. Incinerate sample overnight.
• Weigh cooled ashed sample. 30 minutes.

Total Dietary Fiber:

• Same sample preparation – Total of 5 hours.
• Precipitate with EtOH. 60 minutes.
• Weigh Celite into crucibles, dry for 1 hour, allow to cool. 2 hours
• Prepare crucibles for filtering. 30 minutes
• Filter samples. (can filter 7 samples at once using manifold) Filtering depends on sample. Allow for 2-3 hours.
• Dry crucibles overnight
• Weigh cooled crucibles. 30 minutes.
• Protein determination. 4 hours
• Ash determination. Incinerate sample overnight.
• Weigh cooled ashed sample. 30 minutes.

Schedule (Example):

Total analysis should be complete within 4 days. (Example 7 samples in duplicate + 1 blank)

Day 1: sample incubation and preparation of the crucibles for filtering. Precipitate with IMS overnight.

Day 2: samples and concentrating SDFS filtrate by rotary evaporation to run on HPLC. Crucibles are dried overnight.

Day 3: protein determination and then ashing overnight.

Day 4: collating all the data from HPLC, protein and ash into the MegaCalc report.