Method for simultaneously determining mycophenolic acid ester, mycophenolic acid and metabolite thereof in human blood plasma

A technology of mycophenolate mofetil and mycophenolate mofetil, applied in the field of medical testing, can solve the problems that have not been seen yet, and achieve the effects of simple operation, low cost and high sensitivity

Inactive Publication Date: 2009-04-29
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no domestic and foreign reports on the simultaneou

Method used

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  • Method for simultaneously determining mycophenolic acid ester, mycophenolic acid and metabolite thereof in human blood plasma
  • Method for simultaneously determining mycophenolic acid ester, mycophenolic acid and metabolite thereof in human blood plasma
  • Method for simultaneously determining mycophenolic acid ester, mycophenolic acid and metabolite thereof in human blood plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Volunteer plasma MMF, MPA and MPAG are measured

[0029] Chromatographic conditions

[0030] HPLC system: Japan Shimadzu LC-10A high performance liquid phase instrument (system includes LC-10AD pump, LC-10AT pump, SPD-10A ultraviolet detector, RF-10AXL fluorescence detector, SIL-10A automatic sampler, CBM -10A system communicator, column thermostat, Class-LC10 Version 1.63 chromatographic workstation); chromatographic column: Kromasil-C18 (150mm×4.6mm, 5μm); mobile phase: isocratic elution, methanol——0.1% TFA ( 45:55, V / V); flow rate: 1.2ml / min; column temperature 40°C; fluorescence excitation wavelength 342nm, emission wavelength 425nm.

[0031] Plasma sample pretreatment

[0032] Precisely draw 100 μl of plasma into a 1.5ml centrifuge tube, add 200 μl of acetonitrile solution containing internal standard propafenone 150 μg / ml, vortex for 30 s, centrifuge for 10 min (20,627×g, 4°C), take 20 μl of the supernatant for injection, The internal standard met...

Embodiment 2

[0042] Example 2: Determination of MMF, MPA and MPAG in Plasma of Kidney Transplantation Patients

[0043] Chromatographic conditions

[0044] HPLC system: Japan Shimadzu LC-10A high performance liquid phase instrument (system includes LC-10AD pump, LC-10AT pump, SPD-10A ultraviolet detector, RF-10AXL fluorescence detector, SIL-10A automatic sampler, CBM -10A system communicator, column thermostat, Class-LC10 Version 1.63 chromatographic workstation); chromatographic column: ZORBAX RX-C8 (250mm×4.6mm, 5μm); mobile phase: isocratic elution, methanol——0.1% TFA (52:48, V / V); flow rate: 1.2ml / min; column temperature 45°C; fluorescence excitation wavelength 344nm, emission wavelength 427nm;

[0045] Plasma sample pretreatment

[0046] Precisely draw 100 μl of plasma into a 1.5ml centrifuge tube, add 200 μl of methanol solution containing internal standard propafenone 150 μg / ml, vortex for 30 s, centrifuge for 10 min (20,627×g, 4°C), take 20 μl of the supernatant for injection, T...

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Abstract

The invention belongs to the field of medical test, and relates to a method for analyzing and measuring in vivo drugs, in particular to a method for measuring mycophenolate mofetil, mycophenolic acid and metabolites thereof in human plasma simultaneously. After the pretreatment of a sample to be measured, the method makes use of the characteristic of strong fluorescence absorption of the mycophenolate mofetil and the mycophenolic acid under an alkaline condition, performs derivatization after the analysis of chromatographic column separation, and detects with a fluorescence detector. The method can greatly improve the sensitivity for detecting the mycophenolate mofetil and the mycophenolic acid. The method has the advantages of few samples, simple pretreatment, rapidness, sensitivity, short analysis cycle and low cost, no needs of expensive equipment or reagents, and is suitable for clinical routine monitoring and pharmacokinetic study.

Description

technical field [0001] The invention belongs to the field of medical examination, and relates to an analysis and determination method of drugs in vivo, in particular to a method for determining mycophenolate mofetil, mycophenolic acid and metabolites thereof in human blood plasma. Background technique [0002] Anti-rejection therapy is required after clinical organ transplantation. At present, the new type of anti-metabolism immunosuppressant mycophenolate mofetil (Mycophenolate Mofetil, MMF, RS61443; trade name: Cellcept, Cellcept) is widely used for anti-rejection therapy. The drug is a 2-ethyl ester prodrug of mycophenolic acid (Mycophenolic Acid, MPA), which can significantly increase the bioavailability of MPA in vivo. After oral administration, MMF is rapidly hydrolyzed and deesterified into the active metabolite MPA, which is then combined with glucuronic acid and converted into the inactive metabolite glucuronide (Mycophenolic acid glucuronide, MPAG). like figure 1...

Claims

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Application Information

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IPC IPC(8): G01N33/48G01N30/02G01N21/64
Inventor 道毅俊焦正施孝金李中东钟明康
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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