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OBJECTIVE To established and optimized an analytical method for spiramycin related substances for daily testing of spiramycin related substances. METHODS A Waters Xbridge shield RP18 (4.6 mm×250 mm, 3.5 μm) column was used for the experiment. The chromatographic conditions were as follows: water: 0.2 mol·L-1 dipotassium hydrogen phosphate (adjust the pH value to 9.5 using a 1 mol·L-1 KOH solution): acetonitrile: methanol, (10∶60∶28.5∶1.5) as mobile phase A, water: 0.2 mol·L-1 dipotassium hydrogen phosphate (pH 9.5): acetonitrile: methanol, (10∶30∶57∶3) as mobile phase B, and gradient elution was performed. Detection wavelength 232 nm. RESULTS The established method can achieve good separation between the components of spiramycin and its related substances. The mass concentration of spiramycin Ⅰ is in the range of 0.7-1 200 μg·mL-1, and its peak area shows a good linear relationship with concentration (r=0.999 3). The limit of detection (LOD) is 0.2 μg·mL-1, and the limit of quantification (LOQ) is 0.7 μg·mL-1. The RSD of the repeatability and precision test is less than 2.0%, and the test solution is stable at 5 ℃ for 24 h. CONCLUSION Compared with current analytical methods, this method has strong specificity, good sensitivity and stability, and effectively shortens the testing time and improves the testing efficiency. It can be used for the daily testing of related substances of spiramycin.
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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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