Method for preprocessing biological sample by using freeze drying technique
A biological sample and technical technology, which is applied in the fields of medicine and pharmacy, can solve the problems of many operation steps, long time-consuming, increased sample processing time, etc., and achieve the effect of small particles and large specific surface area
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Embodiment 1
[0023] Embodiment 1: Taking plasma samples as an example
[0024] Separate the plasma from the blood sample to be tested, pipette 0.1ml to 1.5ml into an ependoff tube, put it in a freeze dryer to pre-freeze to -45°C and maintain it for 2 hours, transfer it to a freeze-drying box, turn on the vacuum pump, and wait for 2 hours After the vacuum degree reaches 0.3mbar, heat the material tray and raise the temperature to 10°C according to the predetermined program. The freeze-drying process is completed. After restoring the pressure of the freeze-drying box to atmospheric pressure, take out the freeze-dried plasma sample, and quickly cover the lid of the ependoff tube. After returning to room temperature, 0.1 ml of methanol was added as an extraction solvent, vortexed for 1 minute, and then centrifuged at 12000×g for 10 minutes to obtain the supernatant for liquid phase analysis.
[0025] For example, plasma and tissue homogenate, after freeze-drying, use organic solvents to extrac...
Embodiment 2
[0028] 1) Take the urine and move it into a container, place the container containing the urine sample in the cold trap of the freeze dryer to pre-freeze to -45°C and maintain it for 4 hours, then transfer it from the cold trap to the freeze-drying box, Turn on the vacuum pump, make the vacuum degree below 0.3mbar after 1 hour, and heat the material tray. The first step of heating keeps the temperature of the urine sample lower than the temperature of the eutectic point. After the moisture in the urine sample is removed, the second step of heating is performed. Keep at 20°C for 3 hours, then freeze-drying ends;
[0029] 2) After lyophilization, take out the sample and return to room temperature, add diethyl ether which is 1 / 2 the volume of urine before lyophilization as the extraction solvent, vortex and mix for 1 minute, and centrifuge at 12000×g or more for 10 minutes to take the supernatant for extraction. Correlation analysis tests.
Embodiment 3
[0031] 1) Take the tissue and move it into a container, place the container containing the tissue in the cold trap of the freeze dryer to pre-freeze to -45°C and maintain it for 5 hours, then transfer it from the cold trap to the freeze-drying box, and turn on the vacuum pump After 1 hour, make the vacuum degree below 0.3mbar, and heat the material tray. The first step is to keep the temperature of the tissue lower than the eutectic point temperature. After 5 hours, the freeze-drying can be completed;
[0032] 2) After lyophilization, take out the sample and return to room temperature, add ethyl acetate with tissue volume of 1 or 5 before lyophilization as the extraction solvent, vortex and mix for 2 minutes, centrifuge at 12000×g or more for 20 minutes, and take the supernatant Liquids were analyzed and tested.
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