Development of sustainable method for the Quality Control of traditional medicines on the basis of phytoequivalence and chemical fingerprinting

Abstract

Background: Herbal drugs are composed of single or several types of medicinal plants with additives. This type of preparation is gaining a wide range of popularity among a large a number of people throughout the world. With the increase of usage, it is necessary to maintain the quality of these drug preparations. Therefore, analytical approaches for their intended use in drug quality evaluation need to be validated. This study was planned to develop simple, rapid, selective, precise and economical method for the quality control of herbal preparations. Methods and Materials: Infrared (IR) spectroscopy was used to determine phytoequivalence among the samples. This work outlined a method for identifying herbal drugs on the basis of phytoequivalence. However, as this is a time-consuming and expensive process, the current analysis did not attempt to identify the specific chemical compounds found in the herbs. The marked bands served as a comparative tool for herbal plants and the medications made from them. The quantitative method of comparison was applied for the comparison of crude herbal standard with herbal sample containing formulations. Thin Layer Chromatography (TLC) was used as another analytical method for identification and showing phytoequivalence. It has mostly been used for the qualitative analysis of herbal medicines and to characterize and track the components visually or as an initial separation technique. Single or multiple herbs containing preparation can be analyzed using ultraviolet (UV) spectroscopy. In this technique, it involves the measurement of ultra violet radiation absorption by the substance dissolved in solution. Both qualitative and quantitative analysis can be done through this technique. In the present study, it was aimed at development and validation of UV-spectroscopic technique according to International Conference on Harmonization guidelines which is known as ICH (Q2) guidelines for the analysis of herbs containing polyherbal formulation. The method validation parameters like specificity, precision, accuracy, linearity, range, repeatability and robustness were studied according to ICH (Q2) guidelines. Results and Discussion: Amlaki (Phyllanthus emblica) showed the presence of prominent peak and maximum absorption at 303 nm. The detector response for the Amlaki was linear over the selected concentration range of 1 to 5 μg/mL with a correlation coefficient of 0.998. The absorbance values for intraday precision, found for 1 μg/mL, 3 μg/mL and 5 μg/mL were 0.0082, 0.234 and 0.396 having %RSD of 0.998%, 0.0080% and 0.0058%, respectively. The absorbance values for intermediate precision, found for 1 μg/mL, 3 μg/mL and 5 μg/mL were 0.0084, 0.234 and 0.398 having %RSD 0.6846%, 0.0137% and 0.0038%, respectively. The absorbance value for repeatability was 0.084 having %RSD 0.6901. The accuracy was between 99.348% and 101.478%. Robustness of the method was studied. The %RSD for analyst to analyst variation was 0.4851% and instrument to instrument variation was 0.9726%. The assay results of Amlaki were about 86.588%, 82.150% and 90.828% for three market preparations A, B and C, respectively, indicating insignificant interference from the other ingredients in the formulation. Black plum (Syzygium cumini) showed the presence of prominent peak and maximum absorption at 279 nm. The detector response for the S. cumini was linear over the selected concentration range of 0.1-0.5 μg/mL with a correlation coefficient of 0.9914. The absorbance values for intraday precision found for 0.1 μg/mL, 0.3 μg/mL and 0.5 μg/mL were 0.234, 0.432 and 0.735 having %RSD 0.9245%, 0.5751% and 0.4668%, respectively. The absorbance values for intermediate precision found for 0.1 μg/mL, 0.3 μg/mL and 0.5 μg/mL were 0.233, 0.432 and 0.736 having %RSD 0.8921%, 0.8346% and 0.3421%, respectively. The absorbance value for repeatability was 0.225 having %RSD 0.7950%. The accuracy was between 99.647% and 101.943%. Robustness of the method was studied. The %RSD for analyst to analyst variation was 0.8251% and instrument to instrument variation was 0.3609%. The assay results of S. cumini were about 83.152%, 86.821%, 90.082% and 80.579% for four market preparations A, B, C and D, respectively, indicating insignificant interference from the other ingredients in the formulation. During the method development phase, a number of solvents were used. Among them, methanol was selected in analysis of Amlaki and ethanol was selected in analysis of black plum. These solvents were selected as these solvents satisfied all the conditions relative to peak quality and non-interference at the specified wavelength. The wavelength of maximum absorption (λ max ) was found to be 303 nm and 279 nm in Amlaki and Black plum, respectively. As the values of %RSD is <2%, the method is validated according to ICH (Q2) guidelines. Conclusion: It can be capitulated that this method can be conveniently employed for routine quality control analysis of herbal drugs in bulk drug and other formulations.