Author/Authors :
Moghbel, Abdolhosein School of Pharmacy, Ahwaz University of Medical Sciences, Ahwaz , Rezaee, Saeid School of Pharmacy, Ahwaz University of Medical Sciences, Ahwaz , Zand-Moghaddam, Ahmad School of Paramedicine, Ahwaz University of Medical Sciences , Pedram, Mohamad Shafa Hospital, Ahwaz University of Medical Sciences, Ahwaz,
Abstract :
This article describes a simple and fast high-performance liquid chromatography method for
the determination of methotrexate (MTX) in plasma. Samples were collocted from children
receiving high-dose MTX at shafa Hospital (Ahvaz University of Medical sciences, Ahvaz, iran)
at various times after the end of eachinfusion. Plasma was deproteinized with trichloroacetic
acid and the supernatant was injected into a 250×4.6 mm octadecylsilane column. Mobile phase
was made of TRIS-phosphate buffer (pH 5.7): methanol: acetonitrile (82:11:7) with a flow rate
of 1.8 ml/min. Ultraviolet detection was done at 313 nm and at ambient temperature. Paraaminoacetophenone
was used as internal standard. Methotrexate and internal standard retention
times were 4.4 and 6.5 minutes, respectively. Results showed that reproducibility (precision) of
method within a day was 2.6 to 6 percent and between days was 5.5 to 9.5 percent. The recovery
of the method was between 61.5 and 72.7 percent. The quantitation limit of the method for
methotrexate was 0.1 μM. This method is suitable for quantitation of methotrexate after infusion
of high doses of this drug and has good accuracy, precision and quantiatation limit
