Method for detecting concentration of cycloserine in bone tuberculosis specimens

A technology of cycloserine and bone tuberculosis, which is applied in the field of drug testing, can solve the problems of increased error and long time, and achieve the effect of improving efficiency, reducing error, and eliminating the need to open the cover and weigh.

Active Publication Date: 2020-02-21
BEIJING TUBERCULOSIS & THORACIC TUMOR RES INST +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Method for detecting concentration of cycloserine in bone tuberculosis specimens
  • Method for detecting concentration of cycloserine in bone tuberculosis specimens
  • Method for detecting concentration of cycloserine in bone tuberculosis specimens

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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. The method includes the following steps: (1) the bone tuberculosis specimens are frozenby liquid nitrogen, after the liquid nitrogen is volatilized, a solvent is added for grinding and cycloserine extraction is conducted, and a tissue extraction solution is collected; (2) the tissue extraction solution and a cycloserine standard solution are detected through high performance liquid chromatography-mass spectrometry combination, and during measuring, if the retention time of a chromatographic peak of the tissue extraction solution is consistent with the retention time of a chromatographic peak of the cycloserine standard solution, selected qualitative ions all occur in a sample mass spectrum after background subtraction, and the abundance is consistent compared with the ion abundance of the cycloserine standard solution, and it is judged that the cycloserine is detected; and (3), according to data, obtained in the step (2), of the tissue extraction solution measured through high performance liquid chromatography-mass spectrometry combination, the content of the cycloserinein the bone tuberculosis specimens is quantitatively analyzed through an internal standard method, and thus the concentration of the cycloserine in the bone tuberculosis specimens is obtained. According to the method, the time of processing the bone specimens is shortened, and the measurement concentration is accurate.

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 Applications(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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