Development and validation of UV chromatographic method for quantification of copper and copper nanoparticles in different matrices and pharmaceutical products

Abstract

Background The applications of Cu and CuNPs based on the earth-abundant and inexpensive Cu metal have generated a great deal of interest in recent years, including medical applications. A novel, specific, precise, accurate and sensitive reverse-phase high-performance liquid chromatography (RP-HPLC) method with UV detection has been developed and validated to quantify copper (Cu) and copper nanoparticles (CuNPs) in different biological matrices and pharmaceutical products. Methods The developed method has been validated for linearity, precision, sensitivity, specificity and accuracy. Cu concentration was detected in pharmaceutical products without an extraction process. Moreover, liver, serum and muscle tissues were used as biological matrices. High Cu recovery in biological samples was afforded by using citric acid as a green chelating agent, exact extraction time and pH adjustment. Cu pharmaceutical and biological samples were eluted by acetonitrile: ammonium acetate (50 mM) with 0.5 mg/ml EDTA (30:70 v:v) as an isocratic mobile phase. EDTA reacted with Cu ions forming a Cu-EDTA coloured complex, separated through the C18 column and detected by UV at 310 nm. Results The developed method was specific with a short retention time of 4.95 min. It achieved high recovery from 100.3% to 109.9% in pharmaceutical samples and 96.8–105.7% in biological samples. The precision RSD percentage was less than two. The method was sensitive by achieving low detection limits (DL) and quantification limits (QL). Conclusion The validated method was efficient and economical for detecting Cu and CuNPs by readily available chemicals as EDTA and Citric acid with C18 column, which present the best results on RP-HPLC.

Copper chitosan nanocomposite as antiviral and immune-modulating effect in broiler experimentally infected with chicken anemia virus

“…Liver and muscle tissues (3 chicks per group) were collected on the 1 st , 7 th ,21 st ,35 th , and 42 nd days postmedication to detect copper residue concentration by UV RP-HPLC. Tissue samples were kept at -70 0 C till the analysis (20).…”

“…Agilent C18 column (4.6mm i.d, 150mm, 5µm particle size) was utilized for chromatographic separation. Chromatographic conditions were adjusted (20). Briefly, the UV detector was set at 310 nm.…”

“…The flow rate was 1 ml/min with a specific retention time of 4.955 min. Sample preparation extraction was performed as the former (20). The calibration curve was set as a linear correlation between copper concentration and Area under the Curve (AUC) resulting from spiking blank tissue samples.…”

“…The linear correlation was ranged within eight concentrations 0.1-15 µg/ml of the fortified solution by diluting with 1% nitric acid to ascertain the actual concentration of Cu in Cu-chitosan nanocomposite. The validation process and quantification were calculated, respectively (20,30).…”