Background
Methods
Materials
Plant material and extraction
1H-NMR analysis for metabolite profiling of EECCL
Experimental animals and design
Normal Diet (ND) | High Fat Diet (HFD) | ||
---|---|---|---|
Gold Coin, Malaysia (3.27 kcal/g) | MP Diet, USA (4.39 kcal/g) | ||
Ingredients | Percentage/Amount | Ingredients | Percentage/Amount |
Crude Protein | 21–23% | Casein Purified High Nitrogen | 20.00% |
Crude Fibre | 5.00% | DL-Methionine | 0.30% |
Crude Fat | 3.00% | Sucrose | 30.58% |
Moisture | 3.00% | Corn Starch | 20.00% |
Calcium | 0.8–1.2% | Coconut Oil Hydrogenated | 20.00% |
Phosphorus | 0.8–1.2% | Alphacel, Non-Nutritive Bulk | 5.00% |
Nitrogen Free Extract | 49.00% | DL-a-Tocopherol Powder (250 IU/gm) | 0.12% |
Vitamin A | 10 MIU | AIN-76 Mineral Mix | 4.00% |
Vitamin D3 | 2.5 MIU | MP Vitamin Diet Fortification Mixture 1.2 X Normal Amount | |
Vitamin E | 15.0 g |
Administration of EECCL
Determination of body weight gain, food intake and energy intake
Collection of plasma, liver, lung, kidney, heart, testis, visceral fats, and feces
Determination of fecal fat content
Biochemical measurements
Statistical analysis of data
Results
Metabolite identification of EECCL from 1H-NMR spectra
Metabolite |
1H-NMR signals |
---|---|
Quercetin | δ 6.25 (d, J = 1.5 Hz) |
δ 6.42 (d, J = 1.5 Hz) | |
δ 6.80 (d, J = 1.5 Hz) | |
δ 6.83 (d, J = 8.5 Hz) | |
δ 6.98 (d, J = 8.5 Hz) | |
δ 7.75 (d, J = 2.0 Hz) | |
δ 7.79 (d, J = 2.0 Hz) | |
δ 7.45 (dd, J = 8.5 Hz, 2.0 Hz) | |
Quercetin 3-O-α-rhamnoside | δ 6.25 (d, J = 1.5 Hz) |
δ 6.42 (d, J = 1.5 Hz) | |
δ 6.80 (d, J = 1.5 Hz) | |
δ 6.83 (d, J = 8.5 Hz) | |
δ 7.29 (dd, J = 8.0 Hz, 2.0 Hz) | |
δ 7.75 (d, J = 2.0 Hz) | |
Anomeric proton rhamnosyl | |
δ 4.54 (d, J = 2.0 Hz) | |
Methyl signal; | |
δ 0.92 (d, J = 6.0 Hz) | |
Quercetin 3-O-β-glucoside | δ 6.25 (d, J = 1.5 Hz) |
δ 6.42 (d, J = 1.5 Hz) | |
δ 6.80 (d, J = 1.5 Hz) | |
δ 6.83 (d, J = 8.5 Hz) | |
δ 7.75 (d, J = 2.0 Hz) | |
δ 5.08 (d, J = 7.5 Hz) | |
Anomeric proton glucosyl | |
δ 4.97 (d, J = 7.5 Hz) | |
Rutin | δ 6.25 (d, J = 1.5 Hz) |
δ 6.95 (d, J = 8.5 Hz) | |
δ 7.54 (dd, J = 8.5 Hz, 2.5 Hz) | |
Anomeric proton rhamnosyl | |
δ 4.54 (d, J = 2.0 Hz) | |
Anomeric proton glucosyl | |
δ 4.97 (d, J = 7.5 Hz) | |
Chlorogenic acid | δ 2.07 (m) |
δ 2.22 (m) | |
δ 7.06 (dd, J = 8.5, 1.5 Hz) | |
δ 7.14 (d, J = 1.5 Hz) | |
Signal for caffeoyl | |
δ 6.36 (d, J = 16.0 Hz) | |
δ 7.62 (d, J = 16.0 Hz) | |
Signal for quinic | |
δ 1.91 (d, J = 10.0 Hz) | |
Catechin | δ 2.56 (dd, J = 7.5 Hz, 16.0 Hz) |
δ 2.85 (m) | |
δ 3.91 (m) | |
δ 6.44 (d, J = 2.0 Hz) | |
Epicatechin | δ 6.44 (d, J = 2.0 Hz) |
δ 7.00 (d, J = 8.0 Hz) | |
δ 7.29 (dd, J = 8.0 Hz, 2.0 Hz) | |
δ 4.97 (d, J = 7.5 Hz) | |
Kaempferol | δ 6.26 (d, J = 2.0 Hz) |
δ 6.44 (d, J = 2.0 Hz) | |
δ 6.80 (d, J = 1.5 Hz) | |
δ 7.00 (d, J = 8.0 Hz) | |
δ 8.01 (d, J = 8.5 Hz) | |
Sucrose | δ 4.19 (d, J = 8.5 Hz) |
δ 5.42 (d, J = 3.5 Hz) | |
β glucose | δ 4.60 (d, J = 8.0 Hz) |
α glucose | δ 5.20 (d, J = 3.5 Hz) |
Alanine | δ 1.50 (d, J = 7.5 Hz) |
Valine | δ 1.02 (d, J = 7.0 Hz) |
δ 1.11 (d, J = 6.5 Hz) | |
δ 2.22 (m) | |
Fatty acid | δ 1.35 (m) |
Choline | δ 3.20 (s) |
Isocitric acid | δ 4.12 (d, J = 5.0 Hz) |
Effects of EECCL on body weight gain, visceral fat mass, food and energy intake of rats
Effects of EECCL on organ weights of rats
Dietary group | ||||||
---|---|---|---|---|---|---|
ND | ND + 175 mg/kgBW | ND + 350 mg/kgBW | HFD | HFD + 175 mg/kgBW | HFD + 350 mg/kgBW | |
Liver (g) | 8.66 ± 0.82a
| 8.05 ± 0.42a
| 8.74 ± 1.35a
| 10.4 ± 2.09a
| 9.38 ± 1.05a
| 8.34 ± 0.53a
|
Kidney (g) | 2.22 ± 0.21a
| 2.09 ± 0.33a
| 2.07 ± 0.15a
| 2.19 ± 0.28a
| 2.09 ± 0.23a
| 2.09 ± 0.26a
|
Heart (g) | 1.25 ± 0.17a
| 1.16 ± 0.2a
| 1.16 ± 0.21a
| 1.20 ± 0.13a
| 1.29 ± 0.18a
| 1.17 ± 0.08a
|
Lung (g) | 2.00 ± 0.27a
| 1.79 ± 0.26a
| 1.65 ± 0.34a
| 1.57 ± 0.08a
| 1.82 ± 0.28a
| 1.60 ± 0.28a
|
Testis (g) | 1.55 ± 0.23a
| 1.68 ± 0.28a
| 1.53 ± 0.19a
| 1.54 ± 0.22a
| 1.40 ± 0.45a
| 1.59 ± 0.21a
|
Effects of EECCL on fecal fat content of rats
Effects of EECCL on plasma lipid profiles of rats
Dietary group | ||||||
---|---|---|---|---|---|---|
ND | ND + 175 mg/kg | ND + 350 mg/kg | HFD | HFD + 175 mg/kg | HFD + 350 mg/kg | |
TG | 0.35 ± 0.02c
| 0.32 ± 0.01c
| 0.41 ± 0.08c
| 1.15 ± 0.20a
| 0.70 ± 0.12b
| 0.62 ± 0.09b
|
TC | 1.27 ± 0.17a
| 1.09 ± 0.22ab
| 1.26 ± 0.14a
| 1.28 ± 0.05a
| 1.14 ± 0.06ab
| 1.04 ± 0.05b
|
HDL-c | 0.87 ± 0.14ab
| 0.86 ± 0.11ab
| 0.89 ± 0.08a
| 0.71 ± 0.04b
| 0.72 ± 0.05b
| 0.73 ± 0.03ab
|
LDL-c | 0.25 ± 0.03a
| 0.25 ± 0.05a
| 0.27 ± 0.07a
| 0.26 ± 0.04a
| 0.24 ± 0.04a
| 0.16 ± 0.02b
|
Insulin | 0.21 ± 0.03c
| 0.19 ± 0.02c
| 0.18 ± 0.02c
| 0.35 ± 0.03a
| 0.27 ± 0.06b
| 0.23 ± 0.04bc
|
Leptin | 0.83 ± 0.20bc
| 0.65 ± 0.23c
| 0.68 ± 0.15c
| 2.05 ± 0.38a
| 1.33 ± 0.26b
| 1.12 ± 0.20bc
|
Adiponectin | 17.19 ± 0.95a
| 17.00 ± 2.18a
| 16.11 ± 3.54a
| 10.26 ± 1.92b
| 17.49 ± 2.36a
| 17.07 ± 5.86a
|
Ghrelin | 64.68 ± 8.64ab
| 78.92 ± 26.32a
| 54.78 ± 34.99ab
| 35.25 ± 8.64b
| 89.44 ± 25.94a
| 50.80 ± 7.37ab
|
AST | 93.38 ± 24.24a
| 93.50 ± 11.25a
| 84.85 ± 12.09a
| 156.98 ± 113.32a
| 123.63 ± 32.56a
| 158.90 ± 48.09a
|
ALT | 34.25 ± 4.38bc
| 33.25 ± 6.80c
| 32.08 ± 1.14c
| 51.90 ± 2.46a
| 56.38 ± 7.24a
| 45.95 ± 7.51ab
|
ALP | 80.75 ± 6.02bc
| 79.75 ± 7.59bc
| 66.00 ± 16.85c
| 129.50 ± 23.70a
| 116.30 ± 26.14ab
| 94.00 ± 11.17abc
|
GGT | 1.00 ± 0.00b
| 1.00 ± 0.00b
| 1.25 ± 0.50b
| 3.25 ± 1.26a
| 1.50 ± 1.00b
| 0.75 ± 0.50b
|
Urea | 5.60 ± 0.35a
| 5.58 ± 0.33a
| 5.88 ± 1.07a
| 5.65 ± 0.90a
| 5.08 ± 0.49a
| 5.08 ± 0.54a
|
Creatinine | 48.25 ± 0.96a
| 47.00 ± 3.16a
| 47.75 ± 5.19a
| 52.75 ± 3.30a
| 51.75 ± 2.06a
| 47.25 ± 2.06a
|