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Method for determining human plasma phenytoin and its precursor drug and metabolite

A technology of prodrug and phenytoin, which is applied in the field of medical testing and achieves the effects of strong selectivity, simple operation and less plasma consumption

Inactive Publication Date: 2008-01-16
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant reports on the simultaneous determination of the concentrations of FOS, PHT and 4′-HPPH in human plasma at home and abroad.

Method used

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  • Method for determining human plasma phenytoin and its precursor drug and metabolite
  • Method for determining human plasma phenytoin and its precursor drug and metabolite

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Chromatographic conditions

[0030] HPLC system: column Agilent RX-C 8 (250mm×4.6mm, 5μm), the mobile phase is 0.08% TFA aqueous solution-acetonitrile-methanol (60:17:23, V / V / V), flow rate: 1.5mL min -1 ; Column temperature: 40°C; UV detection wavelength: 210±1nm.

[0031] Plasma sample pretreatment

[0032] Precisely draw 100 μL of plasma sample into a 1.5 mL centrifuge tube, add 50 μL of 20% phosphoric acid for acidification, vortex for 0.5 min, add 1 mL containing 10 mg·L -1 Extract propranolol hydrochloride with ethyl acetate solution, vortex for 5min, centrifuge (15,000×g, 4°C) for 10min, take 800μL of the upper layer extract, transfer to a 5mL centrifuge tube, place in a water bath at 40°C, blow dry with nitrogen , add 100 μL of mobile phase, take 20 μL of supernatant for injection analysis, and quantify by peak area by internal standard method.

[0033] Exclusive

[0034] The blank plasma of 10 subjects who did not take fosphenytoin, phenytoin and 4′-hydroxy...

Embodiment 2

[0042] Chromatographic conditions

[0043] HPLC system: column Agilent RX-C 8(250mm×4.6mm, 5μm), the mobile phase is 0.1% TFA aqueous solution-acetonitrile-methanol (60:17:23, V / V), flow rate: 1.5mL min -1 ;Column temperature: 40°C; UV detection wavelength: 210±1nm; Injection volume: 20μL.

[0044] Plasma sample pretreatment

[0045] Precisely draw 100 μL of plasma sample into a 5 mL centrifuge tube, add 25 μL of 40% phosphoric acid to acidify, vortex for 0.5 min, add 2 mL containing 10 mg L -1 Extract propranolol hydrochloride with ethyl acetate solution, vortex for 5min, centrifuge (15,000×g, 4°C) for 10min, take 1.8mL of the upper layer extract, transfer to a 5mL centrifuge tube, place in a water bath at 40°C, blow with nitrogen After drying, add 100 μL of mobile phase, vortex and redissolve, take 20 μL of supernatant for sample analysis, and quantify by peak area by internal standard method.

[0046] Exclusive

[0047] The blank plasma of 10 subjects who did not take ...

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Abstract

The invention belongs to the medical inspection field, relates to an analysis detection method of drug in the body of a person, and specifically relates to the method that the densities of fosphenytoin, phenytoin and 4'-hydroxyl phenytoin which is the main metabolite of fosphenytoin and phenytoin in can be detected at the same time. The method in the invention is characterized in that pilot sample is pretreated; fosphenytoin, phenytoin and 4'-hydroxyl phenytoin are separated with each other in an acidity flowing phase chromatographic column and are detected by UV detector. The method in the invention has the advantages of little sample, easy, swift and sensitive operation and short analysis period; furthermore, the method in the invention doesn't need expensive equipment and reagent, is suitable for clinical conventional detection and can be used to adjust the drug dose, which can guarantee safe and effective prescription and has the important clinical practice significance.

Description

technical field [0001] The invention belongs to the field of medical testing, and relates to an analysis and determination method for drugs in vivo, in particular to a method for determining fosphenytoin, phenytoin and its metabolite 4'-hydroxyphenytoin in human blood plasma. Background technique [0002] Phenytoin (PHT) is a highly effective drug for the treatment of acute epileptic seizures or status epilepticus. Because of its poor water solubility and strong alkalinity, its use value is affected. Fosphenytoin (FOS), a recently developed parenteral antiepileptic drug (antiepileptic drugs, AEDs), is a phosphate prodrug of PHT. Due to its high polarity and favorable pH, it can be administered by intravenous infusion or intramuscular injection. FOS itself has no obvious pharmacological activity. After administration, it is metabolized into phenytoin by phosphatase in the blood and tissues to exert antiepileptic effect. Phenytoin is mainly metabolized into 4'-hydroxyphenyt...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/74G01N21/33
Inventor 马春来焦正赵倩华洪震
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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