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Method for detecting purithiamine in blood by efficient liquid chromatography

A high-performance liquid chromatography and pyridinethiamine technology, applied in the field of biomedicine, can solve problems such as abnormal thiamine metabolism and thiamine deficiency

Active Publication Date: 2009-09-16
上海兰卫医学检验所股份有限公司
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  • Abstract
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Problems solved by technology

But the questions are: (1) Why are these patients so markedly deficient in thiamine in the absence of dietary deficiency or restriction? (2) In addition to the decrease in the activity of the above three thiamine as coenzymes, why are almost all thiamine metabolic links in AD patients abnormal? (3) Why can't supplementing water-soluble thiamine prevent the occurrence and development of AD? The above problems are obviously difficult to explain by a simple lack of thiamine in the diet

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  • Method for detecting purithiamine in blood by efficient liquid chromatography

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Embodiment 1

[0023] A method for the detection of pyrithione content in blood by high performance liquid chromatography:

[0024] Step 1: Store the blood at -80°C; add potassium ferricyanide to 150g / L sodium hydroxide solution to prepare 0.25g / L potassium ferricyanide solution; add pyrithione to 0.1M hydrochloric acid solution Prepare a 0.1mM pyrithione solution, store it in the dark at 4°C, dilute it with a methanol solution with a weight concentration of 20% on the day of measurement to prepare a 100nM pyrithione solution, set its concentration as Cs; add disodium hydrogen phosphate to the weight concentration Prepare 25mmol / L disodium hydrogen phosphate solution in 10% methanol solution; add sodium dihydrogen phosphate to 70% methanol solution by weight to prepare 25mmol / L sodium dihydrogen phosphate solution;

[0025] The second step: mix the pyridinethiamine solution obtained in the first step with the potassium ferricyanide solution at a volume ratio of 25:3, filter through a 0.2um p...

Embodiment 2

[0030] Another HPLC method for detecting the content of pyrithione in blood:

[0031] Step 1: Store the blood at -80°C; add potassium ferricyanide to 150g / L sodium hydroxide solution to prepare 0.25g / L potassium ferricyanide solution; add pyrithione to 0.1M hydrochloric acid solution Prepare a 0.1mM pyrithione solution, store it in the dark at 4°C, dilute it with a methanol solution with a weight concentration of 20% on the day of measurement to prepare a 100nM pyrithione solution, set its concentration as Cs; add disodium hydrogen phosphate to the weight concentration Prepare 25mmol / L disodium hydrogen phosphate solution in 10% methanol solution; add sodium dihydrogen phosphate to 70% methanol solution by weight to prepare 25mmol / L sodium dihydrogen phosphate solution;

[0032] The second step: mix the pyridinethiamine solution obtained in the first step with the potassium ferricyanide solution at a volume ratio of 25:3, filter through a 0.2um pore size filter membrane to obt...

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Abstract

The invention provides a method for detecting purithiamine in blood by efficient liquid chromatography. The method is characterized in that the method comprises the specific steps: taking whole blood, removing protein, and using the efficient liquid chromatography to detect, wherein the condition of the chromatography comprises: using C18 chromatographic column; taking disodium hydrogen phosphate solution and monosodium orthophosphate obtained in step 1 as a first mobile phase and a second mobile phase respectively; and gradually eluting at the flow speed of 1.0 mL / min, wherein the exciting wavelength of fluoroscopic detection is 385nm and the emitting wavelength is 460nm. The invention has the advantage that the content of the purithiamine in the blood can be detected by a simple method.

Description

technical field [0001] The invention relates to a method for detecting pyrithione in blood by high-performance liquid chromatography, which belongs to the technical field of biomedicine. Background technique [0002] Alzheimer's disease (AD) has become the most common neurodegenerative disease, causing serious harm to individuals, families and society. So far, the pathogenesis of late-onset (sporadic) AD has not been fully resolved. A large number of studies have formed many hypotheses and found some biological targets that are considered to be related to the pathogenesis of AD. Although most of these biological targets are related to β- Amyloid (β-amyloid, Aβ) is related to tau protein metabolism, but it has not been developed into specific diagnostic markers and effective therapeutic measures. The current clinical use of intracerebral cholinesterase inhibitors (such as huperzine A and donepezil, etc.) and non-competitive NMDA receptor antagonists (memantine) cannot preven...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/36C07D401/06
Inventor 钟春玖费国强
Owner 上海兰卫医学检验所股份有限公司
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