A method for detecting cycloserine concentration in bone tuberculosis specimens

A technology of cycloserine and bone tuberculosis, which is applied in the field of drug detection, can solve the problems of long time and increased error, and achieve the effect of improving efficiency, reducing error, and improving biological safety

Active Publication Date: 2022-05-17
BEIJING TUBERCULOSIS & THORACIC TUMOR RES INST +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the relatively long time required for freeze-drying, and two weighings will further increase the error

Method used

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  • A method for detecting cycloserine concentration in bone tuberculosis specimens
  • A method for detecting cycloserine concentration in bone tuberculosis specimens
  • A method for detecting cycloserine concentration in bone tuberculosis specimens

Examples

Experimental program
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Embodiment

[0047] 1. Specimen Processing

[0048] Bone specimen:

[0049] 1) Immediately after taking it out, wash off the attached blood with normal saline, and dry the residual fluid and attached tissue with filter paper;

[0050] 2) After weighing, transfer the bone specimen to a 2mL externally screwed screw tube, first freeze it with liquid nitrogen, and after the liquid nitrogen evaporates, add water according to the ratio of 1g bone specimen weight to 1mL HPLC grade pure water;

[0051] 4) Grinding: Add two ceramic beads in the screw tube, grind on a homogenizer: 6m / s, 100s interval for 1min, repeat once.

[0052] 5) Place the bone homogenate at 4°C for 5 hours to extract the drug (every 1 hour, vortex at a speed of 6m / s for 20s with a homogenizer);

[0053] 6) Centrifugation: 3500g, 4°C, 10min;

[0054] 7) Collect the supernatant and subpackage in EP tubes to obtain the tissue extract (50 μl per tube), to be tested;

[0055] *The broken bone tissue obtained from trauma surgery...

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Abstract

The invention discloses a method for detecting the concentration of cycloserine in bone tuberculosis specimens. It includes the following steps: (1) freeze the bone tuberculosis specimen with liquid nitrogen, after the liquid nitrogen volatilizes, add a solvent to grind and extract cycloserine, and collect the tissue extract; (2) use high-efficiency Liquid chromatography-mass spectrometry is used for detection; when measuring, if the retention time of the chromatographic peak of the tissue extract is consistent with that of the cycloserine standard solution, and in the mass spectrum of the sample after subtracting the background, the selected qualifier ions all appear, and the abundance Consistent with the ion abundance ratio of cycloserine standard solution, it is determined that cycloserine is detected; (3) according to the data of the tissue extract determined by the high performance liquid chromatography-mass spectrometry that step (2) obtains, adopt internal standard method to The content of cycloserine in the bone tuberculosis specimen is quantitatively analyzed to obtain the concentration of cycloserine in the bone tuberculosis specimen. The invention shortens the processing time of the bone specimen and measures the concentration accurately.

Description

technical field [0001] The invention relates to a method for detecting the concentration of cycloserine in bone tuberculosis specimens, belonging to the field of drug detection. Background technique [0002] Bone tuberculosis is a devastating lesion caused by the invasion of Mycobacterium tuberculosis into bones or joints. Most of the diseased sites are in the bones or joints that are heavily loaded, active, and prone to strain. The best site of disease is the spine, followed by hips, knees, feet, elbows, hands, etc. [0003] At present, most of the treatment methods for detecting drug concentration from bone tissue in various studies are freeze-drying at -50°C, weighing the mass, grinding it into powder quickly, and then weighing the required weight. Finally, according to the measured drug concentration, the actual concentration of the drug in the tissue sample was obtained after two conversions based on the dilution ratio of the bone homogenate and the dry-wet mass ratio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N1/28G01N1/42
CPCG01N30/02G01N30/06G01N1/286G01N1/42G01N2030/045G01N2001/2866
Inventor 黄海荣于霞文舒安张婷婷姜广路
Owner BEIJING TUBERCULOSIS & THORACIC TUMOR RES INST
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