Method for detecting content of telbivudine in blood plasma

Abstract

The invention belongs to the field of medical examination and relates to a method for detecting the content of telbivudine in blood plasma by liquid chromatography-tandem mass spectrometry. Accordingto the method, without the need of blow-drying with nitrogen, a sample to be detected directly undergoes acetonitrile dilution/protein precipitation and a supernatant is taken for sample introduction,elution separation is carried out through a mobile phase in a chromatographic column, and detection is finally carried out by a tandem mass spectrometry detector. According to the method, deuterium 3-telbivudine (D3-LdT) is adopted as the internal standard, and a certain amount of acetonitrile precipitated protein is added into a blood sample. After virus inactivation in the blood plasma sample processing, telbivudine is not obviously degraded. Accuracy of the method is guaranteed, and safety of operators is enhanced. By measuring concentration of telbivudine and the internal standard throughthe tandem mass spectrometry, linearity range is within 10-10000 ng/mL, thus meeting requirements of human pharmacokinetics research. Less samples are sampled by the method, pretreatment is simple, fast and sensitive. Only universal equipment and reagents are needed. Analytical period is short, and cost is low. The method of the invention is suitable for the regulation of a hepatitis B patient treatment scheme and monitoring of routine plasma concentration.

Description

technical field [0001] The invention belongs to the technical field of medical testing, and relates to an analysis and determination method for drugs in vivo, in particular to a method for detecting the content of telbivudine in blood plasma by liquid chromatography tandem mass spectrometry, and the liquid chromatography tandem mass spectrometry method is suitable for detecting hepatitis B patients Plasma levels of telbivudine. Background technique [0002] Data show that hepatitis B virus (HBV) infection is a common health problem worldwide. According to WHO reports, about 2 billion people in the world have been infected with HBV, of which 350 million people are chronic HBV infected people, and about 1 million people die every year from liver failure, cirrhosis and liver cancer caused by HBV infection; according to the Ministry of Health of my country in 2006 According to the results of the national hepatitis B epidemiological survey, there are about 93 million chronic HBV ...

AI Technical Summary

Problems solved by technology

[0005] At present, there are two methods for the determination of telbivudine drug concentration in the literature, LC-MS / MS and HPLC-UV, both of which involve the study of telbivudine pharmacokinetics, but the specific detection methods and methodological verification have not yet been seen. detailed report

In addition, the detection methods known from the literature have the following deficiencies or shortcomings: (1) The operation is cumbersome, and the detection methods reported are usually in the sample pretreatment, which needs to be reconstituted after protein precipitation by nitrogen drying and enrichment to improve Sensitivity; (2) Phase I / II clinical trials of telbivudine suggest that the maximum body fluid concentration of telbivudine 600 mg / d is between 0.20 and 6.84 μg / mL, while the linear range of the currently reported detection method is narrower by 0.01 ~5.00μg / mL, in view of the impact of clinical drug combination on blood drug concentration, the current quantitative detection range cannot quickly meet the actual clinical determination needs; (2) The quantitative detection range of HPLC-UV method is 0.1~10.00μg / mL, can not meet the mensuration requirement of telbivudine low concentration point in human plasma while improving detection limit, (3) HPLC-UV method measures telbivudine and the retention time of internal standard is respectively 4.9min and 7.7min, analysis It takes a long time and requires a large amount of mobile phase; (4) the precision is poor; the intraday and interday precision is about 10.6%; (5) because telbivudine is used to treat CHB patients with cross-infection of HBV and HCV , from the perspective of protecting the safety of the measurement operator, the industry recommends that the plasma should be heated to inactivate the virus (58°C, 40min) in the pretreatment, but there are factors affecting the concentration of telbivudine in this operation step, and further research is needed on this. verify

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