3. The efficiency of the lipolysis of triacylglycerides is determined
by measuring the grey intensity of bands corresponding to the
intact triacylglycerides, mono- and diacylglycerides and free
fatty acids at the beginning and end of the digestion protocol
(see Note 15).
4 Notes
1. The meals are designed for a 40–49 years old male, 1.70 m high
and 65 kg weight, 18.5–25 kg/m
2 (body mass index, BMI)
and a moderately activity level (Physical Activity Level,
PAL ¼ 1.70–1.99). With this data the caloric requirement is
2450 kcal.
2. The total daily caloric intake (2450 kcal) is distributed 20–25%
at breakfast, 15–20% at mid-day/snack, 30% at lunch, 25–30%
at dinner.
3. Pasta is cooked in boiling water (1 L per 100 g of pasta) for
12 min. Tuna fish is fried in iron at 120
C for 3 min. Bacon
strips are fried in iron at 120
C for 3 min.
4. This salt is not added to the simulated electrolyte stock solutions as precipitation may occur.
5. Enzymes solutions need to be used on the same day. Weigh the
enzyme carefully because it tends to disperse. The amount of
enzyme to be used for the digestion is recalculated according to
the activity.
6. We measure the α-amylase enzyme activity with an Alphaamylase microplate assay kit.
7. We measure the pepsin enzyme activity with a Pepsin microplate assay kit.
8. Pancreatin is a mixture of several digestive enzymes produced
by the exocrine cells of the porcine pancreas. However, we
measure the lipase enzyme activity to recalculate the amount
of pancreatin required.
9. We measure the total bile acid content in the bile extract with
an assay kit.
10. Particle size distribution of the bile extract may considerably
change among lots as it is depicted in Fig. 2. This may affect the
efficiency of the micellarization. Batch 1 is a fine powder, while
batch 2 should be ground in a mortar to obtain the same
particle size distribution.
11. The pH probe we use in our lab is equipped with temperature
sensor and designed for low conductivity samples. Use cleaning
solution or mild soap or detergent, and a toothbrush to
384
Antonio Pe ´ rez-Ga ´ lvez and Javier Fontecha
by measuring the grey intensity of bands corresponding to the
intact triacylglycerides, mono- and diacylglycerides and free
fatty acids at the beginning and end of the digestion protocol
(see Note 15).
4 Notes
1. The meals are designed for a 40–49 years old male, 1.70 m high
and 65 kg weight, 18.5–25 kg/m
2 (body mass index, BMI)
and a moderately activity level (Physical Activity Level,
PAL ¼ 1.70–1.99). With this data the caloric requirement is
2450 kcal.
2. The total daily caloric intake (2450 kcal) is distributed 20–25%
at breakfast, 15–20% at mid-day/snack, 30% at lunch, 25–30%
at dinner.
3. Pasta is cooked in boiling water (1 L per 100 g of pasta) for
12 min. Tuna fish is fried in iron at 120
C for 3 min. Bacon
strips are fried in iron at 120
C for 3 min.
4. This salt is not added to the simulated electrolyte stock solutions as precipitation may occur.
5. Enzymes solutions need to be used on the same day. Weigh the
enzyme carefully because it tends to disperse. The amount of
enzyme to be used for the digestion is recalculated according to
the activity.
6. We measure the α-amylase enzyme activity with an Alphaamylase microplate assay kit.
7. We measure the pepsin enzyme activity with a Pepsin microplate assay kit.
8. Pancreatin is a mixture of several digestive enzymes produced
by the exocrine cells of the porcine pancreas. However, we
measure the lipase enzyme activity to recalculate the amount
of pancreatin required.
9. We measure the total bile acid content in the bile extract with
an assay kit.
10. Particle size distribution of the bile extract may considerably
change among lots as it is depicted in Fig. 2. This may affect the
efficiency of the micellarization. Batch 1 is a fine powder, while
batch 2 should be ground in a mortar to obtain the same
particle size distribution.
11. The pH probe we use in our lab is equipped with temperature
sensor and designed for low conductivity samples. Use cleaning
solution or mild soap or detergent, and a toothbrush to
384
Antonio Pe ´ rez-Ga ´ lvez and Javier Fontecha