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Hello,
I was wondering if anyone know of a method for the determination of
fluconazole from human plasma. Preferably by HPLC, with a LOQ of ca.
100ng/ml?
Thanks in advance for your help
James Hillis
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James,
We are using the following simple HPLC method:
Calibration curve : 0.05 to 10 mg/L
200 µL plasma sample + 20 µL ei (Pfizer 54373; 100 mg/L) + 10 µL NaOH 2N
Extraction with 4 mL methanol/isopropanol (90/10, V/V)
Injection 20 µL
Column : Novapack C18, 4µm, 150 x 4.6 mm
Mobile phase : Acetate buffer (1g acetate Na + 1 g citric acid, pH 4.5)
/
acetonitrile (350/150, V/V)
Flow : 1 mL/mn
UV detection 260 nm.
Chantal Le Guellec
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Contact Dr. Alan Wu at Hartford Hospital Dept of Clinical Chemistry
hartford CT. It is a GC MS method but he has probably improved that.
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Chantal, thank you for this method. Please can you tell me what volume
the
acetate buffer is made up to. I assume that it is 1000ml.
Thanks
James Hillis
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The volume used is 450 mL. The pH has to be adjusted to 4.5 with NaOH.
Chantal
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