Capillary electrophoresis in-vivo detection method for cobra neurotoxin

Abstract

The invention provides a capillary electrophoresis in-vivo detection method for cobra neurotoxin. The method comprises the following steps of: performing efficient capillary electrophoresis detection on a to-be-detected plasma sample containing FITC-NT (fluorescent single labeled neurotoxin); performing laser induction of a fluorescence detector to detect to obtain an electrophoretogram of the to-be-detected sample; and then comparing the electrophoretogram with a standard curve and calculating concentration of FITC-NT in the to-be-detected sample, wherein the conditions of efficient capillary electrophoresis are as follows: a CEofixTMMEKC kit serves as a buffer liquid, the separating voltage is 20kV, the temperature is 24 DEG C, and the detected wavelengths as follows: the excitation wavelength is 488nm and the emitting wavelength is 520nm. The in-vivo analysis method of FITC-NT, provided by the invention, is high in accuracy, high in precision and good in stability, can quickly and accurately detect neurotoxin in in-vivo blood samples, is high in detection sensitivity and very low in detection limit, can accurately and quantitatively detect a trace amount of FITC-NT components in the blood samples, can be used for in-vivo drug pharmacokinetic research of neurotoxin, and provides accurate data support.

Description

[0001] (1) Technical field [0002] The invention relates to a capillary electrophoresis in vivo detection method of cobra neurotoxin, in particular to a method for detecting cobra neurotoxin by using capillary electrophoresis with high selectivity and high sensitivity combined with a laser-induced fluorescence detector. [0003] (2) Background technology [0004] Cobra neurotoxin (Neurotoxin, NT)——a water-soluble basic polypeptide isolated from the venom of Chinese cobra (Naja naja atra) (C.M.Barber.Alpha neurotoxins[J].Toxicon,2013,66:47–58. ), mainly by binding to the cholinergic receptors in the central brain periaqueductal gray (Periaqueductal Gray, PAG), exerting analgesic effect, its potency is dozens of times higher than that of the same dose of morphine, and there is no gastrointestinal tract Adverse reactions, no tolerance and addiction, is a new type of alternative analgesic drug, mainly used to treat cancer pain and various chronic and intractable pain, etc., and in...

AI Technical Summary

Problems solved by technology

At present, the in vivo detection methods for NT and other protein and polypeptide drugs mainly include immunological analysis, isotope labeling and tracing, and LC-MS / MS. The existing NT detection methods have problems such as safety, accuracy, and cumbersome pretreatment. , to a certain extent, cannot meet the needs of NT in vivo detection, etc., and its application is limited (L.Bailly-Choriberry. Identification ofα-Cobratoxin in Equine Plasma by LC-MS / MS for Doping Control[J].Anal.Chem.,2013,85 :5219–5225)

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