Evaluation of the possibility of individual and joint chromatographic determination of mono- and disaccharides

Purpose of research. To study the conditions of separation and determination of mono- and disaccharides in biological samples and food samples by reverse-phase high-performance liquid chromatography with UV detection. Methods. The studies were carried out using an LC-20AD liquid chromatograph (Shimadzu, Japan) with a spectrophotometric detector, a Supelco LC-18-T column (4.6×250 mm, 5 microns) and LC solution software. An alternative analysis was performed on a KFK-3 spectrophotometer (Russia). Results. The composition of the mobile phase acetonitrile has been experimentally determined for individual and joint determination of glucose, lactose, sucrose and fructose by high-performance chromatography:water is 80:20 and its flow rate is 0.6 ml/min, at which the highest sensitivity of analyte detection is achieved. Calibration functions for the individual determination of carbohydrates and mixtures are constructed, with the individual determination of glucose, lactose, sucrose and fructose, the linear ranges and detection limits (с min ) are 0.5–30.0 mg/ml, с min = 0.15 mg/ml; 25–45 mcg/ml, с min = 9 mcg/ml; 4–16 mg/ml, с min = 1.03 mg/ml; 5–40 mg/ml, с min = 2.35 mg/ml, respectively. When combined: 0.5–15.0 mg/ml, с min = 0.18 mg/ml for glucose; 30–45 mcg/ml, с min = 2 mcg/ml for lactose; 4–32 mg/ml, с min = 2.18 mg/ml for sucrose; 5–30 mg/ml, с min = 3.34 mg/ml for fructose. The greatest sensitivity was recorded in the determination of lactose. The results of the chromatographic analysis were compared with the data of the spectrophotometric determination of carbohydrates, no significant discrepancies were found. Conclusion. Analysis of urine and food samples (milk, syrup, and cookies for diabetics) showed compliance with the analyte content stated by the manufacturer and confirmed the diagnosis of the patient who provided the biological fluid. This allows us to recommend the proposed methods for monitoring the content of mono- and disaccharides in samples with complex matrices.