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To establish a size exclusion high-performance liquid chromatography (SEC-HPLC) method for the determination of the purity of recombinant human basic fibroblast growth factor (rh-bFGF).
The purity of rh-bFGF solution was determined using TSK G2000SWXL column and high-performance liquid chromatography, and the specificity, repeatability, precision, detection limit, stability, and durability of the method were evaluated.
The blank solvent has no interference with the rh-bFGF main peak, and the degradation impurities under various destruction conditions didn't interfere with the determination of the rh-bFGF peak, and the purity index of the rh-bFGF peak was greater than 990. The repeatability and intermediate precision experimental results showed that the separation degree of impurities and the rh-bFGF main peak was greater than 1.5, and the RSD was 0.36% and 0.64%, respectively. The detection limit was 0.185 μg·mL-1. The durability experiment results show that fluctuations in flow rate, column temperature, and wavelength had no effect on the detection results, and the RSD of each condition was less than 2.0%.
The SEC-HPLC method developed has the advantages of simple operation, accurate results, strong specificity, high stability and good applicability, and can be used for rh-bFGF purity determination.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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