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To establish an HPLC method for the determination of six related substances in N-fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine.
The analysis was conducted on YMC Triart C18(250 mm×4.6 mm，3μm) column，the mobile phase was consisted with 0.1% trifluoroacetic acid in water（A） and 0.1% trifluoroacetic acid in acetonitrile（B）at the flow rate of 1.0 mL·min-1. The column temperature was set 30 ℃，the detection wavelength was 265 nm and the injection volume was 10 μL.
N-Fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine had good separation from the adjacent impurity peaks；The resolution of impurity1- 6 was greater than 1.5，and showed a good linear relationship (r≥0.999) in the corresponding mass concentration range，the detection limit of impurity 1-6 was 0.03 μg·mL-1，and the quantitative limit was 0.06 μg·mL-1，the average recovery rate (n=9) of impurity 1-6 was in the range of 97.6%-98.8%. The results of the three batches of N-fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine showed that the contents of impurity 2，impurity 4 were <0.2% and <0.1%，respectively，and the other four impurities were not detected，and content of the total impurity was <1%.
This method has good resolution，high sensitivity and strong specificity，and is suitable for the determination of related substances in N-fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine.
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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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