Background
Methods
Herbal materials
Remedies | Method of preparation and administration | Traditional uses |
---|---|---|
THP-R001a | Pounded, dried, mixed with honey and made into boluses | refreshing and rejuvenating effect on the entire body; reduces joint and muscle pain and flatulence |
Take one bolus twice a day before meals | ||
THP-R002a | Pound, mixed with boiled water or honey and made into boluses; take 4–5 boluses thrice a day before meals | nourishes the skin; promotes blood circulation and used as a blood tonic |
THP-R003a | Pounded, dried, mixed with honey and made into boluses; take 2–4 boluses twice a day before meals | health tonic and promotes longevity |
THP-R004a | Pounded, dried, mixed with honey and made into boluses; take one bolus once a day before meals | health tonic, skin refreshment, and promotes longevity |
THP-R005b | Dried and boiled with water; drink 40–50 mL thrice a day before meals | carminative and a tonic |
THP-R006b | Pounded, dried, mixed with honey and made into boluses; take two boluses twice a day after meals | stimulating and rejuvenating the entire body |
THP-R007b | Pounded and infused in hot water (90–97 °C, 3–5 min); drink 100–150 mL of the infusion twice a day after meals | health promoting and hypolipidemic agents |
THP-R008c | Dried, boiled with water, and filtered the infusion; drink 50–60 mL twice a day after meals | used as a health tonic and used to treat fatigue |
THP-R009c | Dried, boiled with water, and filtered the infusion or pounded, dried, mixed with honey and made into boluses; drink 50–60 mL or take two boluses twice a day after meals | health tonic, sex stimulant, or sexual performance enhancer |
THP-R010d | Extracted by boiling water for at least 3 h; drink 40–50 mL thrice a day before meals | an appetite inducer and tonic |
THP-R011e | Pounded, dried, and made tinctures (alcohol extract) or mixed with honey and made into boluses; drink 30 mL or take three boluses once a day before meals | nourishing and tonifying agents; reduces joint and muscle pain, and applied as a sex stimulant |
THP-R012f | Dried, boiled with water, and filtered the infusion; drink 40–50 mL thrice a day before meals | improvement blood circulation tonic |
THP-R013g | Extracted by boiling water for at least 3 h; drink 10–20 mL thrice a day before meals | stimulates and rejuvenates the entire body |
THP-R014h | Dried, boiled with water, and filtered the infusion; drink 5 mL thrice a day before meals | health tonic and sex stimulant; reduces joint and muscle pain and lowers blood glucose |
THP-R015h | Extracted by boiling water for 15 min; drink 5 mL thrice a day before meals | tonic, sexual performance enhancer, and lowering blood glucose |
THP-R016h | Pounded, dried, and mixed approximately 5 g with 100 mL of warm water; drink once a day before meals | stimulate and rejuvenate the entire body |
THP-R017i | Pounded, dried, and mixed approximately 5 g with 100 mL of warm water; drink once a day before meals | blood tonic, reduce joint and muscle pain |
THP-R018j | Extracted by boiling water for at least 3 h and filtered the infusion; drink 50 mL thrice a day before meals | stimulate and rejuvenate the entire body |
THP-R019k | Pounded, dried, extracted by boiling in water, and filtered the infusion; drink 50 mL once a day before meals | blood tonic; reduce joint and muscle pain |
THP-R020k | Pounded, dried, and made tinctures (water extract); drink 50 mL once a day before meals | health tonic |
Preparation of polyherbal extracts
Quantification of phytochemical contents
Total phenolic content (TPC)
Total flavonoid content (TFC)
Gas chromatography-mass spectrometry analysis of essential oils from some herbal ingredients of an effective formula, THP-R016
Metal-chelating activity (MCA)
Antioxidant properties
DPPH and ABTS radical scavenging assays
Ferric-reducing antioxidant power (FRAP) assay
Superoxide anion radical scavenging assay
Peroxyl radical scavenging assay
Cytotoxic effects
Statistical analysis
Results
Antioxidant related phytochemicals
Samples | Phenolic content | Flavonoid content |
---|---|---|
mg gallic acid/g extract TPC | mg catechin/g extract TFC | |
THP-R001 | 39.3 ± 5.3 | 37.7 ± 1.8 |
THP-R002 | 134.1 ± 6.3 | 63.4 ± 2.3 |
THP-R003 | 114.1 ± 7.1 | 51.6 ± 0.9 |
THP-R004 | 151.9 ± 4.6 | 68.4 ± 1.0 |
THP-R005 | 107.7 ± 4.8 | 60.8 ± 0.2 |
THP-R006 | 77.3 ± 4.6 | 41.9 ± 0.1 |
THP-R007 | 93.8 ± 2.7 | 43.4 ± 0.1 |
THP-R008 | 109.7 ± 6.8 | 55.3 ± 2.3 |
THP-R009 | 72.5 ± 3.1 | 40.1 ± 0.3 |
THP-R010 | 269.8 ± 11.1 | 131.2 ± 1.5 |
THP-R011 | 106.9 ± 5.1 | 64.7 ± 2.0 |
THP-R012 | 150.7 ± 7.5 | 80.9 ± 0.8 |
THP-R013 | 81.7 ± 6.1 | 61.3 ± 3.1 |
THP-R014 | 207.2 ± 8.2 | 103.0 ± 3.1 |
THP-R015 | 211.0 ± 2.9 | 42.9 ± 0.9 |
THP-R016 | 384.6 ± 3.1 | 51.9 ± 0.7 |
THP-R017 | 148.0 ± 9.5 | 132.7 ± 1.5 |
THP-R018 | 62.1 ± 3.4 | 37.0 ± 0.2 |
THP-R019 | 347.4 ± 12.0 | 225.3 ± 2.4 |
THP-R020 | 57.0 ± 3.4 | 34.2 ± 1.5 |
Ability of extracts to act as preventive and chain-breaking antioxidants
Samples | Yields | MCAa | FRAPb assay | Radical scavenging activities; IC50 (mg/mL) | |
---|---|---|---|---|---|
% (w/w) | IC50 (mg/mL) | μM Fe2SO4/mg extract | DPPH | ABTS | |
THP-R001 | 5.68 | 0.93 ± 0.00 | 11.61 ± 0.25 | – | – |
THP-R002 | 4.60 | 0.94 ± 0.01 | 39.79 ± 0.72 | 2.15 ± 0.41 | 1.15 ± 0.00 |
THP-R003 | 5.68 | 0.63 ± 0.02 | 18.58 ± 0.35 | – | 2.01 ± 0.03 |
THP-R004 | 3.99 | 0.80 ± 0.01 | 42.51 ± 0.76 | 0.92 ± 0.02 | 1.09 ± 0.06 |
THP-R005 | 10.54 | 0.84 ± 0.01 | 38.84 ± 0.78 | 1.61 ± 0.03 | 1.38 ± 0.04 |
THP-R006 | 8.93 | 0.64 ± 0.01 | 15.63 ± 0.30 | – | 1.96 ± 0.10 |
THP-R007 | 8.07 | 0.64 ± 0.01 | 20.86 ± 0.65 | – | 1.45 ± 0.05 |
THP-R008 | 10.95 | 0.88 ± 0.01 | 51.62 ± 0.46 | 1.77 ± 0.05 | 1.31 ± 0.03 |
THP-R009 | 11.83 | 0.30 ± 0.01 | 20.27 ± 0.46 | 2.42 ± 0.05 | 1.79 ± 0.02 |
THP-R010 | 8.26 | 2.81 ± 0.04 | 87.12 ± 1.15 | 0.53 ± 0.02 | 0.42 ± 0.00 |
THP-R011 | 8.52 | 0.58 ± 0.02 | 30.33 ± 0.65 | 2.10 ± 0.03 | 1.31 ± 0.01 |
THP-R012 | 9.04 | 1.46 ± 0.06 | 40.70 ± 0.95 | 1.04 ± 0.07 | 0.87 ± 0.01 |
THP-R013 | 10.07 | 0.49 ± 0.01 | 20.92 ± 0.27 | 2.22 ± 0.04 | 1.89 ± 0.05 |
THP-R014 | 2.67 | 0.58 ± 0.02 | 54.55 ± 1.24 | 1.43 ± 0.07 | 0.55 ± 0.01 |
THP-R015 | 5.52 | 1.21 ± 0.01 | 84.22 ± 1.23 | 0.43 ± 0.02 | 0.70 ± 0.01 |
THP-R016 | 10.96 | 1.27 ± 0.02 | 95.76 ± 2.35 | 0.16 ± 0.00 | 0.34 ± 0.00 |
THP-R017 | 4.60 | 0.58 ± 0.02 | 82.32 ± 0.81 | 1.51 ± 0.24 | 1.07 ± 0.01 |
THP-R018 | 3.13 | 0.95 ± 0.02 | 18.75 ± 0.42 | – | 2.46 ± 0.10 |
THP-R019 | 4.31 | 1.24 ± 0.03 | 96.86 ± 1.63 | 0.50 ± 0.02 | 0.40 ± 0.01 |
THP-R020 | 4.09 | 0.57 ± 0.01 | 18.57 ± 0.15 | – | – |
Trolox | – | – | – | 0.09 ± 0.01 | 0.34 ± 0.01 |
EDTA (μg/mL) | – | 8.48 ± 0.34 | – | – | – |
Correlation between free radical inhibition activity and phenolic compounds
Pearson’s correlation (p value) | |||||
---|---|---|---|---|---|
ABTSa | FRAPa | MCAa | TPCb | TFCb | |
DPPHa | 0.959 (0.000) | 0.884 (0.000) | −0.690 (0.001) | 0.882 (0.000) | 0.592 (0.006) |
ABTS | 0.897(0.000) | −0.711 (0.000) | 0.939 (0.000) | 0.695 (0.001) | |
FRAP | −0.734 (0.000) | 0.895 (0.000) | 0.699 (0.001) | ||
MCA | −0.751 (0.000) | −0.449(0.047) | |||
TPC | 0.682 (0.001) |
Inhibitory activity of extracts on superoxide anion radicals and cytotoxic effects
Peroxyl radical scavenging activity of an effective remedy, THP-R016
Medicinal plants | Yield | RT | Compound names | Match Factor | Formula | MW | %Peak area |
---|---|---|---|---|---|---|---|
Alpinia galanga L. | 0.43 | 20.72 | β-Bisabolene | 95.7 | C15H24 | 204 | 14.0 |
18.91 | cis-α-Bergamotene | 96.6 | C15H24 | 204 | 10.0 | ||
21.06 | β-Sesquiphellandrene | 94.0 | C15H24 | 204 | 9.7 | ||
8.08 | 1,8-Cineole | 97.3 | C10H18O | 154 | 6.0 | ||
16.55 | Chavicol, acetate | 98.8 | C11H12O2 | 176 | 4.3 | ||
Cyperus rotundus L. | 1.02 | 21.31 | 4-Isopropyl-1-methoxy-1,6-dimethyl- −1,2,3,4-tetrahydronaphthalene | 90.3 | C15H22 | 202 | 11.7 |
17.52 | Copaene | 95.2 | C15H24 | 204 | 5.0 | ||
16.81 | α-Cubebene | 94.5 | C15H24 | 204 | 4.7 | ||
21.69 | β-Calacorene | 89.9 | C15H20 | 200 | 4.5 | ||
21.61 | 11-hydroxy-2,3,4,11-tetrahydro-6H-pyrimido[2,1-b]quinazolin-6-one | 71.5 | C11H11N3O2 | 217 | 3.9 | ||
Piper retrofractum Vahl. | 0.87 | 18.59 | Caryophyllene | 92.1 | C15H24 | 204 | 9.4 |
20.52 | Pentadecane | 96.7 | C15H32 | 212 | 7.4 | ||
19.42 | 1,4,7,-Cycloundecatriene, 1,5,9,9-tetramethyl-, Z,Z,Z- | 95.0 | C15H24 | 204 | 6.8 | ||
24.78 | 8-Heptadecene | 96.0 | C17H34 | 238 | 6.8 | ||
25.06 | Heptadecane | 96.6 | C17H36 | 240 | 6.1 |