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.