Background
Methods
Materials and reagents
Sample preparation
Preparation of EPHF
Preparation of the tested sample
Preparation of the standard solution
Identification of compounds by LC-MS
Determination of compounds by HPLC
Pharmacological activity of EPHF
Determination of antioxidant capacity
Determination of cell growth inhibition activity
Determination of AChE inhibitory activity
Results and discussion
The preparation of EPHF
Qualitative and quantitative analysis of EPHF
Optimization of the sample preparation
Qualitative analysis of EPHF
Method validation for quantitative analysis
Compound | Calibration curve |
r
| Linear range/μg | LOQ/ng | LOD/ng |
---|---|---|---|---|---|
Ideain |
Y = 5000000x-5992.1 | 0.9999 | 0.0049 ~ 0.63 | 4.93 | 2.47 |
Cyanidin 3-arabinoside |
Y = 5000000x-12627 | 0.9999 | 0.0049 ~ 0.63 | 4.89 | 2.44 |
Hyperoside |
Y = 3000000x-70021 | 0.9998 | 0.0049 ~ 5.02 | 4.90 | 2.45 |
Isoquercitrin |
Y = 3000000x-68963 | 0.9997 | 0.0049 ~ 5.01 | 4.89 | 2.45 |
(−)-Epicatechin |
Y = 835115x-7850.7 | 0.9999 | 0.0099 ~ 5.12 | 9.99 | 2.50 |
Chlorogenic acid |
Y = 4000000x-131307.7 | 0.9995 | 0.0050 ~ 2.57 | 5.01 | 2.50 |
Compound | Precision RSD (%) | Reproducibility | Recovery | ||
---|---|---|---|---|---|
Intra-day | Inter-day | RSD (%) | Average (%) | RSD (%) | |
Ideain | 1.45 | 1.45 | 1.38 | 98.39 | 3.60 |
Cyanidin 3-arabinoside | 1.53 | 1.44 | 1.74 | 102.78 | 1.92 |
Hyperoside | 1.46 | 1.93 | 2.12 | 100.68 | 0.73 |
Isoquercitrin | 1.44 | 1.81 | 0.81 | 103.05 | 1.49 |
(−)-Epicatechin | 1.36 | 1.92 | 1.72 | 101.51 | 3.15 |
Chlorogenic acid | 1.76 | 1.72 | 0.34 | 99.05 | 0.99 |
Quantitative analysis of samples
Samples | Ideain | Cyanidin 3-arabinoside | Hyperoside | Isoquercitrin | (−)-Epicatechin | Chlorogenic acid | Total |
---|---|---|---|---|---|---|---|
EPHF | 17.94 | 0.14 | 0.04 | 0.02 | 4.09 | 1.00 | 23.23 |
PHF | 0.087 | 0.010 | 0.038 | 0.022 | 0.64 | 0.084 | 0.881 |
Pharmacological activity of EPHF
Antioxidant capacity of EPHF
AChE inhibitory activity of EPHF
HF | PHF | EPHF | Galanthamine | |
---|---|---|---|---|
Anti-AChE (IC50, μg/mL) | 1385 | 616.40 | 11.72 | 0.29 |