Method for measuring chemical components in urine and excrement of rat orally taking relinqing granules
A technology of hot drenching and feces clearing, which is applied in the field of research on the metabolism of Chinese patent medicines, and can solve problems such as the lack of description of the content determination method
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Embodiment 1
[0175] Method for determining the chemical composition in rat urine after oral administration of Relinqing Granules: Take 0.5 g of Relinqing Granules and dissolve them in 5 times of water to make an aqueous solution with a concentration of 0.1 g / mL, and administer them to rats by intragastric administration (245g) 2.5mL aqueous solution of hot shower clear granules, collect urine for 0-2h, centrifuge the urine in a low-speed centrifuge at 4000r / min for 10min, transfer the upper layer liquid to a new blank EP tube, press with ethyl acetate Extract at a volume ratio of 1:4 for 5 minutes, vortex evenly for 30 seconds, then centrifuge at a constant temperature at 4°C at a speed of 12,000r / min for 10 minutes, transfer the supernatant to a new EP tube, repeat 3 times, and blow at 40°C in a nitrogen blower. After drying, the residue was dissolved in 100 μL of mobile phase, vortexed for 60 seconds, then 12000 r / min, kept at 4°C, centrifuged for 10 minutes, and 10 μL of sample was taken...
Embodiment 2
[0182] Method for determining the chemical composition in rat urine after oral administration of Relinqing Granules: take 1 g of Relinqing Granules and dissolve them in 8 times of water to make an aqueous solution, and give rats (265 g) Relinqing Granules aqueous solution by intragastric administration 2.65mL, collect 10-12h of urine, centrifuge the urine in a low-speed centrifuge at 4000r / min for 10min, transfer the upper layer liquid to a new blank EP tube, extract with ethyl acetate at a volume ratio of 1:4 for 5min, Vortex evenly for 30s, then centrifuge at constant temperature at 4°C, rotate at 12000r / min, centrifuge for 10min, transfer the supernatant to a new EP tube, repeat 3 times, blow dry at 40°C with a nitrogen blower, and wash the residue with 100μL mobile phase Dissolve, vortex and mix for 60 seconds, then 12000r / min, 4°C constant temperature, centrifuge for 10 minutes, take 10μL sample, and treat the blank urine in the same way, using liquid chromatography-mass s...
Embodiment 3
[0189] Method for determining the chemical composition in rat urine after oral administration of Relinqing Granules: Take 4g of Relinqing Granules and dissolve them in 10 times of water to make an aqueous solution, and give rats (273g) Relinqing Granules aqueous solution by intragastric administration 2.73mL, collect 24-48h of urine, centrifuge the urine in a low-speed centrifuge at 4000r / min for 10min, transfer the upper layer liquid to a new blank EP tube, extract with ethyl acetate at a volume ratio of 1:4 for 5min, Vortex evenly for 30s, then centrifuge at constant temperature at 4°C, rotate at 12000r / min, centrifuge for 10min, transfer the supernatant to a new EP tube, repeat 3 times, blow dry at 40°C with a nitrogen blower, and wash the residue with 100μL mobile phase Dissolve, vortex and mix for 60 seconds, then 12000r / min, 4°C constant temperature, centrifuge for 10 minutes, take 10μL sample, and treat the blank urine in the same way, using liquid chromatography-mass sp...
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