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Method for measuring content of 5-fluorouracil in plasma and colorectal cancer cells based on high performance liquid chromatography

A high-performance liquid chromatography, cancer cell technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of affecting accuracy, large interference, low drug content, etc., to improve peak shape, optimize chromatographic conditions, and improve recovery. rate effect

Inactive Publication Date: 2016-05-18
JIANGSU INST OF NUCLEAR MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although it has been reported in the literature that HPLC is used to determine 5-Fu, the absorption of 5-Fu in the small intestine is poor, the oral bioavailability is low, the drug content in cells and plasma is extremely low, and the interference of endogenous substances in cells and plasma is large, and the extraction The recovery rate of the method is low, and it is difficult to achieve the required detection sensitivity by conventional determination methods, thus affecting the accuracy of 5-Fu determination in biological samples

Method used

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  • Method for measuring content of 5-fluorouracil in plasma and colorectal cancer cells based on high performance liquid chromatography
  • Method for measuring content of 5-fluorouracil in plasma and colorectal cancer cells based on high performance liquid chromatography
  • Method for measuring content of 5-fluorouracil in plasma and colorectal cancer cells based on high performance liquid chromatography

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Biological sample pretreatment

[0053] Plasma sample processing

[0054] Activate the solid-phase extraction column StyreScreen?HP with 1mL ethanol and 1mL deionized water (containing 0.1% trifluoroacetic acid in mass concentration) sequentially; absorb 0.5mL plasma sample, add it to the solid-phase extraction column at a speed of 1mL / min, wait until the sample is enriched After collection, rinse with 1mL deionized water and drain; elute with ethanol-deionized water containing 0.1% trifluoroacetic acid (V / V, 75︰25) solution, and drain; use a vacuum centrifugal evaporation concentrator to dry for 20min , remove the organic solvent; dry and concentrate in a freeze dryer, and dilute to an appropriate concentration with mobile phase during determination;

[0055] Colorectal cancer cell (SW480-B7 and SW480-NC) sample processing

[0056] The solid-phase extraction column StyreScreen?HP was activated with 1mL ethanol and 1mL deionized water (containing 0.1% triflu...

Embodiment 2

[0058] Embodiment 2 chromatographic condition optimization

[0059] 1. Selection of extraction column (comparison of different extraction methods)

[0060] ODS extraction column, StyreScreen? HP solid-phase extraction column and the common ethyl acetate extraction method and ether-isopropanol extraction method reported in the literature were used respectively. The column was first activated according to the above method, and the preparation containing 5-Fu1.0, 4.0, 12.0μg / mL low, medium and high concentrations of plasma samples and colorectal cancer cells samples were processed by different extraction methods, and the content of 5-Fu was determined by high performance liquid chromatography.

[0061] Extraction efficiency=measured value of C / standard value of C×100%. Using StyreScreen? HP extraction, the efficiency is greater than 85%, higher than ordinary organic solvent extraction.

[0062] The results of the extraction efficiency are shown in Table 1.

[0063] Table 1 Mea...

Embodiment 3

[0073] Embodiment 3 standard curve and detection limit:

[0074] plasma standard curve

[0075] Take an appropriate amount of 5-Fu standard substance dissolved in water, make 1mg / mL standard solution, dilute to 500, 100, 50, 10, 5, 1, 0.5μg / mL, take 100μL respectively, add healthy person’s unadministered Dilute the blank plasma to 2mL, mix well, configure the standard blood drug concentration of 25, 5, 2.5, 0.5, 0.25, 0.05, 0.025μg / mL, perform HPLC determination after solid phase extraction according to the above plasma sample processing method , record the peak area; take the 5-Fu concentration as the abscissa, and the corresponding peak area as the ordinate to perform linear regression, and the linearity is: Y=34783x+34562, R 2 =0.9991, the results showed that the mass concentration of 5-Fu showed a good linear relationship between 0.025~25μg / mL. According to the signal-to-noise ratio (S / N) of 3, the minimum detection concentration of 5-Fu is 0.02 μg / mL.

[0076] Colorect...

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Abstract

The invention relates to a method for measuring content of 5-fluorouracil in plasma and colorectal cancer cells based on high performance liquid chromatography and belongs to the technical field of liquid chromatography medical analysis. 5-fluorouracil (5-Fu) is a miazines antineoplastic drug widely applied clinically at present; the antineoplastic chemotherapy effect of 5-Fu is related to the concentration of 5-Fu in the cancer cells; other side effects are caused when the drug concentration in the body is ultrahigh. For the purpose of enhancing the drug therapeutic effect and reducing the side effects, the trend of the concentration of 5-Fu in the cancer cells and the drug concentration in human body shall be strictly monitored. The monitoring has significant clinical significance in promotion of drug therapeutic effect, reduction of toxicity and prevention of post-operation transfer. According to the method provided by the invention, 5-Fu in plasma and colorectal cancer cells is extracted in the manner of solid phase extraction, so that the extraction efficiency is increased; the chromatographic condition is optimized; the peak pattern is improved by adding a trifluoroacetic acid and methyl alcohol system into a detected flow phase; the number of theoretical plates is increased and the chromatographic condition is mild; under a selected optimal chromatographic condition, the measurement for 5-Fu is more precise and accurate than the measurement in literature reports.

Description

technical field [0001] The invention relates to a high-performance liquid chromatography method for measuring 5-fluorouracil content in blood plasma and colorectal cancer cells, belonging to the technical field of liquid chromatography medical analysis. Background technique [0002] 5-Fluorouracil (5-Fu) is currently the most widely used pyrimidine antineoplastic drug in clinical practice, and is often used in the treatment of malignant tumors such as colorectal cancer, liver cancer, and gastric cancer. However, clinically different tumor patients have obvious individual differences in the efficacy and toxicity of 5-Fu. Its anti-cancer chemotherapy effect has a great relationship with the drug concentration in cancer cells, and other side effects will occur when the drug concentration in the body is too high. In order to enhance the curative effect of drugs and reduce adverse side effects, its concentration in cancer cells and the dynamics of drug concentration in the body ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/88G01N30/06G01N2030/062G01N2030/8822
Inventor 周杏琴毛师师邹美芬徐希杰
Owner JIANGSU INST OF NUCLEAR MEDICINE
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