ТHE VALIDATION CHARACTERISTICS STUDY OF THE TECHNIQUE OF DETERMINING IMPURITIES OF URSODEOXYCHOLIC ACID IN SOLID FORMS BY THE THIN-LAYER CHROMATOGRAPHY METHOD
Keywords:
ursodeoxycholic acid, hard gelatin capsules, analytical technique, thin-layer chromatography, validation characteristics.Abstract
Ursodeoxycholic acid (UDCA) is one of the native bile acids, which is synthesized in the process of normal bile acid metabolism in the human body and is widely used in the form of solid dosage forms as a first-line therapeutic agent for various cholestatic diseases and hepatobiliary pathologies of the liver and the treatment of cholesterol gallstones. During the synthesis, impurities such as related bile acids, namely chenodeoxycholic acid (impurity A), cholic acid (impurity B) and lithocholic acid (impurity C), which are used as starting material, as well as other organic compounds, such as aldehydes and ketones, may enter the UDCA substance. We have developed an analytical technique for determining UDCA impurities in hard gelatin capsules by thin-layer chromatography, which is characterized by expressiveness and relatively low cost of routine control compared to liquid chromatography. The formed spots of identified impurities were evaluated in comparison with the spots of standard solutions. The analysis results were considered reliable if two spots were observed in the chromatogram of the reference solution (g). The validation characteristics of specificity, robustness, suitability of the chromatographic system, and detection limit were evaluated. Robustness was investigated using an unsaturated chamber and it was proven that the method is resistant to changes in the degree of saturation of the chambers. The suitability of the chromatographic system was tested on plates of the same manufacturer with aluminum and glass substrate types. It was established that the type of sorbent substrate does not affect the suitability of the chromatographic system. The maximum permissible values of the detection limits for UDCA and its impurities were calculated and it was proven that the spots of impurities at the detection limit level are clearly observed.
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