Citation: LUO Ning, LIU Pinghuai, WU Xiaona, CHEN Chen, ZHANG Ling, HE Yifei. Determination of ethylene glycol bis(2-aminoethyl) ether-N,N,N,N-tetraacetic acid in medicine using high performance liquid chromatography[J]. Chinese Journal of Chromatography, ;2015, 33(4): 419-422. doi: 10.3724/SP.J.1123.2014.11014 shu

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

1.
Key Study Center of the National Ministry of Education for Tropical Resources Utilization/Ministry of Education Key Laboratory of Protection, Development and Utilization of Tropical Crop Germplasm Resources, Hainan University, Haikou 570228, China

Corresponding author: LIU Pinghuai,
Received Date: 12 November 2014
Available Online: 3 February 2015
Fund Project: 国家科技支撑计划项目(2011BAD14B01) (2011BAD14B01)海南省中药现代化科技专项(ZY201327) (ZY201327)国家科技型中小企业技术创新基金项目(13C26244604892). (13C26244604892)

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×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 ℃. 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.
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