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

A technology of mycophenolate mofetil and mycophenolic acid, which is applied in the field of medical testing, can solve problems that have not yet been seen, and achieve the effect 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

[0003] So far, there have been no domestic and foreign reports on the simulta

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Volunteer urine sample MMF, MPA and MPAG are measured

[0026] Chromatographic conditions

[0027] 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 oven, 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;

[0028] Urine Sample Pretreatment

[0029] Precisely draw 100 μL of urine sample into a 1.5 mL centrifuge tube, add 500 μL of methanol solution to dilute, vortex for 15 s, centrifuge for 5 min (20,627×g, 4°C), take 20 μL of supernatant for injection, and quantify by peak area by external standard method.

[0030] Exclus...

Embodiment 2

[0037] Example 2: Determination of urine sample MMF, MPA and MPAG in kidney transplant patients

[0038] Chromatographic conditions

[0039] 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;

[0040] Urine Sample Pretreatment

[0041] Precisely draw 100 μL of urine sample into a 1.5 mL centrifuge tube, add 500 μL of methanol solution to dilute, vortex for 15 s, centrifuge for 5 min (20,627×g, 4°C), take 20 μL of supernatant for injection, and quantify by peak area by external standa...

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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 urine 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, no needs of expensive equipment or reagents, short analysis cycle and low cost, 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 simultaneously determining mycophenolate mofetil, mycophenolic acid and metabolites thereof in human urine. Background technique [0002] Mycophenolate Mofetil (MMF, RS61443; trade name: Cellcept, Xiaoxi) is a new type of antimetabolite immunosuppressant, which is widely used clinically for anti-rejection therapy after organ transplantation. 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). Such as figure 1 shown. The concentration of MPAG in the body is much...

Claims

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Application Information

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