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Method for stabilizing carbidopa/levodopa in biological matrix

A biological matrix, levodopa technology, applied in the biological field, can solve problems such as the inability to truly reflect the drug situation, and achieve the effects of reducing enzymatic reactions, reducing enzymatic reactions, and inhibiting oxidative reactions

Active Publication Date: 2021-09-17
苏州华测生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous data reflect that CD / LD has stability problems in biological matrices such as whole blood and plasma. If it cannot be resolved, the test data in the experiment cannot truly reflect the situation of the drug in the body.

Method used

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  • Method for stabilizing carbidopa/levodopa in biological matrix
  • Method for stabilizing carbidopa/levodopa in biological matrix
  • Method for stabilizing carbidopa/levodopa in biological matrix

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Measure 9mL of sterile water for injection to completely dissolve, then add 1000μL of formic acid to obtain 10% FA as a stabilizer;

[0025] Add 25 μL of stabilizer 10% FA to the pre-cooled EP tube, then add 0.975 mL of plasma, mix well, add 100 μL of 0.03 mg / mL CD / LD, mix again and divide into 3 parts in parallel. The first sample was taken immediately for quantitative detection, the second sample was placed in wet ice environment for 3 hours before quantitative detection, and the third sample was placed under wet ice environment for 20 hours before quantitative detection. See Table 1 and Table 2 for the results.

Embodiment 2

[0027] Measure 8mL of sterile water for injection to dissolve completely, then add 2000μL of formic acid to obtain 20% FA as a stabilizer;

[0028] Add 25 μL stabilizer 20% FA to the pre-cooled EP tube, then add 0.975 mL plasma, mix well, add 100 μL 0.03 mg / mL CD / LD, mix again and divide into 3 parts in parallel. The first sample was taken immediately for quantitative detection, the second sample was placed in wet ice environment for 3 hours before quantitative detection, and the third sample was placed under wet ice environment for 20 hours before quantitative detection. See Table 1 and Table 2 for the results.

Embodiment 3

[0030] Measure 9.5mL sterile water for injection, then add 500μL trifluoroacetic acid to obtain stabilizer 5% TFA;

[0031] Add 25 μL of stabilizer 5% TFA to the pre-cooled EP tube, then add 0.975 mL of plasma, mix well, add 100 μL of 0.03 mg / mL CD / LD, mix again and divide into 3 parts in parallel. The first sample was taken immediately for quantitative detection, the second sample was placed in wet ice environment for 3 hours before quantitative detection, and the third sample was placed under wet ice environment for 20 hours before quantitative detection. See Table 1 and Table 2 for the results.

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Abstract

The invention discloses a method for stabilizing carbidopa / levodopa in a biological matrix, a stabilizer is added into the biological matrix, and the stabilizer comprises an acidifying reagent with the volume fraction of 5% and an antioxidant with the mass fraction of 10%. On the basis of adjusting the pH value and adding the antioxidant, the purpose of protecting the CD / LD in the biological matrix is achieved, and finally the biological matrix sample containing the CD / LD can be stored in various environments. By using the method, a whole blood sample containing CD / LD can be preserved for 2 hours under the condition of 2-8 DEG C, a plasma sample can be preserved for 20 hours under the condition of 2-8 DEG C and can be preserved for 30 days under the condition of lower than -60 DEG C, and 4 times of freeze-thaw cycles can be performed. The requirements for collection, transportation, storage and detection of samples in CD / LD pre-clinical / clinical tests are met, and a basis is provided for pharmacokinetic research of CD / LD.

Description

technical field [0001] The invention relates to a method for stabilizing carbidopa / levodopa in a biological matrix, in particular to a method for meeting the collection, transportation, storage and detection requirements of samples in carbidopa / levodopa preclinical / clinical trials The method belongs to the field of biotechnology. Background technique [0002] Levodopa (Levodopa, LD) is the most effective drug for the treatment of Parkinson's disease (PD), and it is a prodrug of dopamine (Dopamine, DA). It can pass through the blood-brain barrier and be decarboxylated into DA by Dopa decarboxylase (DDC) in the brain to play a role. Since DDC not only exists in the brain, but also widely exists in other organs and blood vessel wall cells. If LD is used alone, only about 1% of the drug can enter the central nervous system. Therefore, LD combined with peripheral dopa decarboxylase inhibitors ( Dopadecarboxylase inhibitor (DDCI) such as carbidopa (Carbidopa, CD) or benserazide ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0231A01N1/0226
Inventor 冷明红叶双双陆国才夏玉叶宗英胡明文叶爱华刘大伟
Owner 苏州华测生物技术有限公司
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