Acetamiprid development of moderate conditions of high-performance liquid chromatography method in the separation of biological fluids from blood and urine.

The analysis (Shimadzu Corporation) was carried out on a high-performance liquid chromatograph SPD–M20A, equipped with a model LC-20 Prominence (US) diode matrix UV-detector autosamplers. The process consists of a four-channel gradient type pump designed for high-pressure operation, a spectrophotometric detector with a wavelength of 190–360nm, and a measuring device with a volume of 20μL – “Rheodyne” injector and a chromatographic column. The research was conducted under the following conditions:

– Perfect Sil 300 ODS C18 with chromatographic steel column particle size 5μm. Filled with the sorbent, size 150×4.6mm;

– excited phase acetonitrile-water 70:30;

– reduction phase flow -0.75ml/min;

– Sample size for analysis – 20mkl;

– detector wavelength 246nm;

– analysis time 10minutes.

Acetamiprid was weighed to 0.01g (a.t), dissolved in a 50mL volumetric flask with a mobile phase, and the volume of the solution was delivered by the mobile phase to the mark. From this solution, standard working solutions of acetamiprid were prepared and the separation time of acetamiprid was determined.

As a result of the analysis, the retention time of acetamiprid on the chromatogram was 2.314minutes in the blood and 2.722minutes in the urine. The obtained results were determined by metrological analysis with 67.56% separation of acetamiprid from model blood samples with a relative error of 1.63%, with a relative error of 73.98% with acetamiprid of the model urine sample.

The retention time of acetamiprid pesticide in the standard sample under the conditions developed by the HPLC method was 2,190minutes. Chromatographic peaks were formed in the test blood sample, which lasted 2,314minutes, and 2.722minutes in the urine. The amount of acetamiprid isolated from model blood and urine samples was 67.56% in the blood and 73.28% in the urine during metrological analysis.

Moderate conditions for the detection of acetamiprid by high-efficiency liquid chromatography have been developed. On the chromatogram, acetamiprid was identified by chromatographic peaks with a retention time of 2,190minutes in standard samples. When the samples tested were also chromatographed, values equal to the retention time of standard acetamiprid were recorded. Under the conditions of the developed analysis, positive results were obtained and 67.56% of biological fluids were isolated from blood samples and 73.28% from urine samples. The developed methods of analysis were applied to the detection of biological fluids, and pesticides isolated from them.