Validation of a simple HPLC method to quantify methotrexate concentrations in human plasma

Original Research

Abstract

Methotrexate (MTX) is a chemotherapy and immunosuppressive agent widely used to treat cancer, autoimmune diseases in children and adult patients, and ectopic pregnancy. However, MTX is highly toxic to the liver, kidney, and nervous system. This study aimed to quantify the concentration of MTX in human plasma using high-performance liquid chromatography (HPLC). MTX and its internal standard (para aminoacetophenone-PAPA) in plasma samples were extracted simultaneously with methanol. Sample purity was performed using the 1 cc OASIS HLB cartridges. Sample injection volume of 10 µL was analyzed on a Lichrocart Supersil 125-4 column C18 maintained at 40 °C on a Waters 2695 XE equipped with a PDA detector set at 303 nm. The mobile phase contained phosphate buffer (pH 6.0) and methanol at a ratio of 80:20 (v/v) and was maintained at a flow rate of 1 ml/min. The results showed that the total time of chromatographic analysis was 15 min. MTX and PAAP were found in the chromatograms at retention times of 2.3 and 5.2 min, respectively. The linear range of the MTX from 0.5 to 25 µg/mL. Intra-day and inter-day imprecision for MTX ranged from 3.42 to 8.128%. LLOQ of MTX was 0.5 µg/mL and the extraction effects were above 77%. In conclusion, we developed and validated a simple HPLC method to determine the MTX concentrations in human plasma.
 

Graphical abstract

Identification of Curcuma aromatica growing in Vietnam and its potential anticancer components

Original Research

Abstract

Curcuma aromatica, the herbal medicine belongs to Zingiberaceae family, is well known for anti-tumor activity through multiple pathways and a potential candidate for complementary medicine in cancer treatment. The aims of this study were to distinguish between Curcuma species based on polymorphisms of the nucleotide sequence of chloroplast DNA (cpDNA) and preliminarily analyze their potential-anticancer compounds. Totally six samples supposed C. aromatica growing in An Giang province, Vietnam were collected. The contents of curcumin, curdione, and germacrone in the six samples were analyzed and compared by using the high-performance liquid chromatography (HPLC) method. All specimens were identified according to their trnSfMintergenic spacer sequences by Sanger sequencing. Among the six samples, three were determined as C. aromatica, two were C. longa, and one was C. zedoaria. Curcumin, curdione, and germacrone, known as anticancer compounds, were simultaneously found in sample NT3 that identified as C. aromatica by Sanger sequencing. The obtained results revealed a potential herbal candidate for complementary and alternative medicine.

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