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Method for preprocessing biological sample by using freeze drying technique

A biological sample and technology technology, applied in the fields of medicine and pharmacy, can solve the problems of many operation steps, long time consumption, increased sample processing time, etc., and achieve the effect of small particles and large specific surface area.

Inactive Publication Date: 2011-06-22
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] In the process of basic research or clinical drug concentration monitoring, there are often a large number of samples to be tested. Using the traditional sample pretreatment method, the endogenous components in the sample are likely to interfere with the determination of the target drug, and there are many operating steps. , especially when the drug to be tested is a thermally unstable compound, the processing process needs to be carried out at low temperature or room temperature, which will inevitably increase the time required for sample processing
[0013] Traditional biological sample pretreatment methods have disadvantages such as many operation steps, long time-consuming, and many endogenous interferences.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to a method for preprocessing a biological sample by using freeze drying technique. In the invention, a freezedryer is utilized to make the biological sample into freeze-dried powder, organic solvent is utilized to extract target compounds in the sample, and the target compounds are analyzed. The method completely eliminates moisture in the sample, reduces interference of endogenous substance, changes the sample to be loose porous powder, increases the specific surface of the sample, enhances the extraction percentage of the organic solvent to the target compound and simplifies the operating procedure. Because the freezedryer can process a plurality of samples at the same time, analysis and test procedure to a plurality of biological samples is simplified, and the time for preprocessing the samples can be saved.

Description

technical field [0001] The invention belongs to the fields of medicine and pharmacy, and in particular relates to a method for pretreatment of biological samples by using freeze-drying technology. Background technique [0002] Freeze vacuum drying is also called drying. Sublimation drying or lyophilization for short. It is one of the drying methods, the purpose is to store items. [0003] The reason why items are damaged, rotten, and deteriorated is mainly caused by two factors: external factors and internal factors. The external factors are the effects of air, water, temperature, organisms, etc.; If the effects of external and internal factors can be minimized, the object can be kept unchanged for a certain period of time. [0004] The drying method is to get rid of the moisture contained inside the article, because moisture is one of the necessary conditions for the growth of all organisms. When the water content of the organism is reduced to a certain extent, the orga...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/42F26B5/06
Inventor 杨广德张继业曾爱国宋鑫
Owner XI AN JIAOTONG UNIV
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