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. 

1. Tereshchuk L.V., Starovoitova K.V. Food chemistry. Kemerovo: Kemerov. gos. un-t; 2020. 126 p. (In Russ.)

2. Dedov I.I., Shestakova M.V., Vikulova O.K. Epidemiology of diabetes mellitus in the Russian Federation: clinical and statistical analysis according to the Federal register of diabetes mellitus. Sakharnyi diabet = Diabetes mellitus. 2017;20(1):13-41. (In Russ.) https://doi.org/10.14341/DM8664

3. Arbatsky A.P., Traktina E.P., Smirnova L.A. Investigation of chemical properties of chitosan by titrimetric method. Zhurnal prikladnoi khimii = Journal of Applied Chemistry. 2006;79(2):272-274. (In Russ.)

4. Bavykina T.Yu. Study of desorption of easily digestible carbohydrates from the oral cavity. Actual problems of medicine. 2023;46(1):73-78. https://doi.org/10.52575/2687-0940-2023-46-1-73-78.

5. Turdalieva P.K. Tribulus terrestrisl. Carbohydrates. Universum: khimiya i biologiya = Universum: Chemistry and Biology. 2024;2-1(116):58-60. https://doi.org/10.32743/UniChem.2024.116.2.16718.

6. Kropacheva T.N. Electrochemical determination of carbohydrates in wines using copper electrodes. Vestnik of Technological University = Vestnik of Technological University. 2023;26(7):32-36. https://doi.org/10.55421/1998-7072_2023_26_7_32.

7. Kartsova L.A. Capillary electrophoresis method: biomedical applications. Laboratory and Production. 2023;2(24):32-46. https://doi.org/10.32757/2619-0923. 2023.2.24.32.46.

8. Kartsova L.A., Bessonova E.A., Somova V.D. Hydrophilic interaction chromatography. Zhurnal analiticheskoi khimii = Journal of Analytical Chemistry. 2019;74(5):415-424. https://doi.org/10.1134/ S1061934819050058. (In Russ.)

9. Moskvin L.N., Kostanyan A.E., Moskvin A.L., Rodionov O.V., Yakimova N.M. Methods for the continuous chromatographic separation of substances. Zhurnal analiticheskoi khimii = Journal of Analytical Chemistry. 2023;78(6):671-680. (In Russ.) https://doi.org/10.1134/s1061934823040111.

10. Dmitrienko S.G., Apyari V.V., Tolmacheva V.V., Gorbunova M.V. Liquid-liquid extraction of organic compounds into a single drop of the extractant: overview of reviews. Zhurnal analiticheskoi khimii = Journal of Analytical Chemistry. 2021;76(8):907-919. (In Russ.) https://doi.org/.1134/S1061934821080049.

11. Rybal'chenko I.V., Baigil'diev T.M., Rodin I.A. Chromatography–mass spectrometry analysis for the determination of the markers and biomarkers of chemical warfare agents. Zhurnal analiticheskoi khimii = Journal of Analytical Chemistry. 2021;76(1):26-40. (In Russ.) https://doi.org/10.1134/ S1061934821010111.

12. Bošnir J., Bevardi M., Hećimović I., Budeč M., Juranović Cindrić I., et al. Optimization of sample preparation procedure for determination of fat-soluble vitamins in milk and infant food by HPLC technique. Processes. 2024;12(7):1530. https://doi.org/10.20944/preprints202403.0889.v1.

13. Temerdashev Z.A., Abakumov A.G., Shelud'ko O.N., Yakuba Yu.F., Tsypko T.G. Chromatographic methods in the identification and determination of the component composition and quality of wines. Journal of Analytical Chemistry. 2024;79(10):1364-1386. (In Russ.) https://doi.org/10.1134/S1061934824700734.

14. Deineka V.I., Oleinits E.Yu., Blinova I.P., Deineka L.A. Replacement of acetonitrile with ethanol in the determination of anthocyanins by reversed-phase high-performance liquid chromatography. Journal of Analytical Chemistry. 2023;78(1):63-67. (In Russ.) https://doi.org/10.1134/s1061934822110053.

15. Melikhova E.V., Egorov G.A., Lazutkina E.I., Elfimova A.V. Optimization of conditions for the determination of folic acid by spectrophotometric, fluorimetric chromatographic analysis. Izvestiya Yugo-Zapadnogo gosudarstvennogo universiteta. Seriya: Tekhnika i tekhnologii = Proceedings of the Southwest State University. Series: Engineering and Technology. 2022;12(3):184-200. (In Russ.) https://doi.org/10.21869/2223-1528-2022-12-3-184-200.

16. Korenman Ya.I., Mokshina N.Ya., Bychkova A.A. Physico-chemical determination of carbohydrates in food and beverages. Vestnik Voronezhskogo gosudarstvennogo universiteta inzhenernykh tekhnologii = Bulletin of the Voronezh State University of Engineering Technologies. 2014;(1):146-152. (In Russ.)

17. Zenkevich I.G., Deruish A. Analytical aspects of the dependence of the retention indices of organic compounds in reversed-phase HPLC on the content of methanol in the eluent. Analitika i kontrol' = Analytics and Control. 2022;26(1):41-48. (In Russ.) https://doi.org/10.15826/analitika.2022.26.1.00.

18. Liu J., Li J., Yi D., Liu Y., Liu R., Xue Y., et al. Non-derivatization strategy for the comprehensive characterization of neutral monosaccharide isomers and neutral disaccharide isomers using hydrophilic interaction liquid chromatography coupled to quadrupole/time-of-flight mass spectrometry. Journal Chromatography B: Analalytical Technologies in the Biomedical Life Sciences. 2021;1185:122972. https://doi.org/10.1016/j.jchromb.2021.122972

19. Kartsova L.A., Malyushevskaya A.V., Kolobova E.A. Analytical capabilities of the determination of carbohydrates determination by chromatographic and electrophoretic methods. Journal of Analytical Chemistry. 2023;78(2):144-161. (In Russ.) https://doi.org/10.31857/S1061934823020041.