Detecting method for determining various neurotransmitters on the basis of in situ derivation

A neurotransmitter and detection method technology, applied in the field of analytical chemistry, can solve problems such as long reaction time, low sensitivity, matrix interference, etc., achieve good recovery results, improve sensitivity, and good recovery effects

Active Publication Date: 2016-08-17
山东住融物业有限公司
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  • Abstract
  • Description
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Benefits of technology

This patented describes a new type of chemical substance called Neuromethylinium-2Hydroxyproline(NM 2 HPRT). These compounds have several advantages such as high sensing capabilities, ability to identify different types of neuron transmiters within one minute without being affected by other factors that affect their signal quality. They also improve upon resolution and precision during mass spectral measurements while reducing environmental concerns associated therewith. Overall these improvements make it possible to efficiently and effectively analyze large amounts of samples from both human subjects and animals.

Problems solved by technology

This patented technical problem addressed by this patent relates to developing an improved method for detecting small amounts of compounds called neuropeptors during neuroscience researches without causing damage from other materials present within them. Current methods involve complicated procedures involving expensive equipment like gaschrommetry and liquid nitrogen analyzers.

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  • Detecting method for determining various neurotransmitters on the basis of in situ derivation
  • Detecting method for determining various neurotransmitters on the basis of in situ derivation
  • Detecting method for determining various neurotransmitters on the basis of in situ derivation

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Experimental program
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Embodiment 1

[0037] For the analysis method of neurotransmitter standard in brain microdialysate of Parkinson's disease and Alzheimer's disease rats:

[0038] Neurotransmitter standards (19 kinds, purchased from Sigma Reagent Company) were prepared in acetonitrile / water to obtain a concentration of 5.0 × 10 −5 mol L -1 neurotransmitter standard solution. Take 50 µL of neurotransmitter standard solution and place it in a 1.5 mL centrifuge tube, add 30 µL of internal standard solution to the standard solution (internal standard solution consists of 10 µL with a concentration of 1×10 -5 mol / L isoproterenol solution in acetonitrile, 10 µL concentration is 1×10 -5 mol / L 5-oxindole-2-carboxylic acid solution in acetonitrile and 10 µL concentration is 1×10 -5 mol / L norleucine in acetonitrile), adding 800 µL of NaHCO with pH 9.5 3 -Na 2 CO 3 Buffer solution, then immediately add 80 µL bromobenzene and 200 µL 4'-carbonyl chloride-rhodamine-acetonitrile solution with a molar concentration ...

Embodiment 2

[0044] 1. Rat grouping, drug administration and brain microdialysis sampling:

[0045] Sixteen male SD rats weighing 200-250 g were divided into two groups, the first group was the control group, and the second group was the model group. The modeling process and behavioral evaluation of Parkinson's model were consistent with the literature (Pharmacology Biochemistry and Behavior, 2011, 98: 286-291). The above-mentioned model group rats need to go through the following three experimental stages: the first stage, after successful modeling and free-moving Parkinson’s disease model rats, free to drink water, eat and drink, and automatically collect brain microdialysate in an awake state , and the above analysis method was used to determine the content of neurotransmitters; in the second stage, the rats in the first stage were given baicalein (280 mg·kg -1 ) for treatment, and automatically collect the brain microdialysate; when the concentration of neurotransmitters returns to cl...

Embodiment 3

[0052] 1. Alzheimer's disease model rats and urine sampling:

[0053] Eight male SD rats with a body weight of 200-250 g need to go through the following two experimental stages: the first stage is for rats in normal state, free to drink water, eat and drink, and collect their urine to detect the concentration of neurotransmitters; In the second experimental stage, the modeling process and behavioral evaluation of Alzheimer's disease model rats were carried out with reference to the literature (Journal of Mass Spectrometry, 2015, 50: 354-363), which was consistent with the literature, indicating that the modeling was successful. Drink water, eat and drink freely, and collect their urine.

[0054] Determination of neurotransmitter content in brain microdialysate by ultrasonic-assisted-in situ derivatization-dispersive liquid-liquid microextraction combined with ultra-high performance liquid chromatography triple quadrupole mass spectrometry:

[0055] Take 50 µL of urine from t...

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Abstract

The invention belongs to the field of analytical chemistry and particularly relates to a detecting method for determining various neurotransmitters on the basis of in situ derivation. The method is characterized in that 4'-carbonyl chloride-rhodamine is used as the derivative reagent for the first time, the ultrasonic assistance-in situ derivation-dispersive liquid-liquid microextraction technology is used to derive amino acid and monoamine neurotransmitters, and the ultra-high performance liquid chromatography triple quadrupoles tandem mass spectrum with multiple reaction monitoring modes is used to perform qualitative and quantitative detection; three internal standard substances (isoprenaline, 5-ketoindole-2-carboxylic acid and norleucine) are used for building the linear equations of the neurotransmitters of three chemical structures; the existence of the neurotransmitters is accurately determined by quantification through an internal standard method. The detecting method is simple, fast, high in sensitivity, good in selectivity, good in recovery rate, capable of fast and accurately determining the neurotransmitters, and the like.

Description

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Claims

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Application Information

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Owner 山东住融物业有限公司
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