Determination of ethylene glycol bis (2-aminoethyl) ether-N,N,N,N-tetraacetic acid in medicine using high performance liquid chromatography

Abstract

A method was developed for the determination of ethylene glycol bis(2-aminoethyl) ether-N,N,N,N-tetraacetic acid (EGTA) by high performance liquid chromatography (HPLC). The content of EGTA can be determined by that of EGTA-Cu through the complexation between EGTA and Cu2+. The chromatographic separation was performed on an Ultimate-AQ C18 analytical column (250 mm x 4.6 mm, 5 µm) using the mobile phase of acetonitrile-ion-pairing reagents (with 0.3% tetrabutyl ammonium hydroxide in mass fraction adjusted to pH 6.50 using acetate)-buffered saline (35 mmol/L sodium acetate of pH 6.50) (20:20:60, v/v/v). The chromatographic conditions were as follows: flow rate, 1.50 mL/min; detection wavelength, 245 nm; injection volume, 100 µL; column temperature, 40 °C. Under the conditions, good linear relationships between the mass concentration and the peak area of EGTA were observed in the range of 0.10-15.00 mg/L (R = 0.9998). The limit of detection (LOD, S/N = 3) and limit of quantitation (LOQ, S/N = 10) were determined as 0.05 mg/L and 0.17 mg/L, respectively. The average recoveries were 98.34%-99.03% with the RSDs of 1.08%-3.33% (n = 9). The results showed that the developed method is sensitive, accurate, reproducible and suitable for the analysis of EGTA in medicine.