PHARMACEUTICAL ANALYSIS OF THE SUBSTANCE LACTITOL BY HPLC
DOI:
https://doi.org/10.32689/2663-0672-2025-2-27Keywords:
lactitol, disaccharide, HPLC, chemical and thermal degradation of the molecule, pharmaceutical analysis, accompanying impurities, substanceAbstract
Lactitol is used in medical and pharmaceutical practices as an active pharmaceutical ingredient (API) or excipient (sweetener, texturizer) in prebiotic complexes. The State Pharmacopoeia of Ukraine (SPU) does not regulate the analysis of the substance lactitol. The European Pharmacopoeia (Eur.Ph.) regulates the analysis of lactitol in the form of monohydrate and imposes certain requirements for the pharmaceutical analysis of this substance. The identification of lactitol monohydrate is recommended to be carried out by IR absorption spectrophotometry, thin layer chromatography (TLC) and liquid chromatography (LC) methods (2.2.29). In order to increase the efficiency and effectiveness of pharmaceutical analysis of the lactitol substance, we have introduced a more modern high-performance liquid chromatography (HPLC) method with higher identification capability.The purpose of the work. To adapt an alternative chromatography method – a high-tech method of high-performance liquid chromatography (HPLC) in the analysis of the test substance lactitol in order to detect unacceptable impurities of various origins in its composition; adapt chromatography conditions and modify research methods to ensure protection of lactitol substance molecules from different types of degradation.Methodology. The disaccharide lactitol (4-O-α-D-galactopyranosyl-D-glucitol) is a carbohydrate alcohol derived from lactose (milk sugar). According to the recommendations of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), lactitol is classified as safe. The Food and Drug Administration (FDA, USA) has approved the use of lactitol in food products (GRAS). Lactitol is used in the manufacture of sugar-free foods (ice cream, chocolate, candy, baked goods, frozen fish, pasta, chewing gum, infant formula). In the European Union, lactitol has a mark E966. A positive property of lactitol is that it does not increase blood sugar levels, as it is not absorbed in the intestines. This makes it possible to use it in the manufacture of products for diabetics. The SPU does not regulate the analysis of lactitol substance. Eur.Ph. recommends the analysis of lactitol monohydrate using chromatographic methods such as TLC and LC. However, as practice shows, the HPLC method provides a more thorough analysis of substances with the detection of accompanying impurities not regulated by Pharmacopoeias and allows an objective conclusion to be drawn regarding the degree of purity of the tested substance.Scientific novelty. Introduction into the practice of pharmaceutical analysis of the substance lactitol of the modern HPLC method by adapting the chromatography conditions developed for the LC method and modifying the research methods with optimal conditions for protecting molecules from chemical and thermal degradation.Materials and methods. Samples of lactitol substance, pharmacopoeial standard samples of SPU lactitol monohydrate; HPLC, Agilent 1200 chromatograph with refractometric detector, SUPELCOGEL Ca chromatographyc column, column temperature 60˚С-85˚С; flow – 0.6 ml/min; injection volume – 10 μl; chromatography time – 60 min; for the determination of impurities by HPLC, the following reagents were used: acetonitrile R (HPLC grade), water for chromatography R (HPLC grade), methanol R (HPLC grade), glycerol R (HPLC grade); computer analysis using the OpenLab CDS program.Conclusions. The conditions for HPLC chromatography of the lactitol substance were adapted to determine its purity and detect the stability of its molecules against degradation under changing research conditions. A system of mobile phases with modification of the composition and ratio of their components was proposed: mobile phase (1): H2O – CH3CN (12:88, V/V), 28.87 ml; mobile phase (2): H2O – CH3OH –CH3CN (10:5:85, V/V/V), 21.48 ml; mobile phase (3): H2O – CH3CN (20:80, V/V), 13.28 ml).Chromatographically, the formation of products of chemical and thermal degradation of lactitol – D-galactose (Rt=11.228) and D-sorbitol (Rt=28.050) was not confirmed. An accompanying impurity of glycerin – an excipient in pharmaceutical compositions of lactitol – was detected. The retention time and plane of analytical signals (peaks) of the test substance lactitol change significantly when using modified mobile phases: for lactitol from 6.800 min to 7.182 min (Rt), for glycerol from 9.533 min to 15.786 min (Rt) and when varying the column temperature from 60 to 85°C.
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