Homeopathy is a therapeutic system based on three fundamental principles: the similarity (similia similibus curantur), the infinitesimal dose (in decimals of Hahnemann, XH, or in centesimals of Hahnemann, CH) and the individualization of the method (symptomatology, medical and life history, physical and psychological state). Homeopathic medicinal products are made by sequentially energized ultra dilutions of ‘strains’. The treatment given to the patient causes in a healthy person the symptoms that will neutralize in a sick person. Compounds are prepared systematically, diluting the active substance in water and alcohol, stirring vigorously between dilutions. The active substance may be of plant, mineral or animal origin.
Since its inception homeopathy has been subject to discussion among scientists and experts in the areas of medicine and pharmacology, but the use of knowledge is already a reality in many countries, both developed and underdeveloped. According to the current situation, the main concern is in improving the quality of pharmacological therapy, this is the common interest for health authorities, professionals, patients and for the whole society.
The efficacy and safety of these treatments are linked to the physicochemical and compositional characteristics of the species active in the dilutions. UV-visible spectrophotometry is an economical, efficient and fast analytical technique to characterize solutions regardless of the dilution of the same or the nature of the sample. An ideal tool to analyze the difference between the properties of solutions that are extremely dilute, such as homeopathic. It is based on the fact that the molecules absorb the electromagnetic radiation that affects the material, and in turn that the amount of light absorbed depends linearly on the concentration. Homeopathic drugs require a longer wavelength than other substances because they are in an ultra-dilute state. The use of this analysis technique has generated relevant contributions to the process of characterization of this type of compounds.
Aspects to consider when using the UV-visible spectrophotometer
- Standardize the method of analysis: consists of verifying and documenting the degree of security that allows the strict traceability of the results. Accurate and accurate data, which are within the specifications and quality attributes required to be used as treatment. The method must detect the presence of emitted errors, the criteria to counteract can be statistical.
- Linearity and range: these are characteristics to be evaluated from the equipment to be used for characterization. It is important to know the ranges in which the concentration limits of the analytical method lose its linearity. It is recommended to make use of statistical methods for its evaluation, measuring a minimum of four samples.
- Precision: This parameter expresses the degree of dispersion between a series of multiple take measurements due to random errors inherent in the test method.
- Accuracy: This characteristic depends on the sample matrix, the preparation procedure and the concentration of the analyte in the sample. Excess occurs when there is interference and the selectivity of the method is not adequate, and by default suggests that the matrix is complex and the extraction of the analyte requires a more extensive process.
- Detection Limit: Defined as the minimum amount of analyte in the sample that the computer can detect.
- Limit of quantification of the method: is the minimum amount of analyte present in the sample that can be quantified under experimental conditions.
- Sensitivity: Defined by the ability to distinguish between different concentrations, it measures the instrument’s response factor as a function of concentration. The concentration response is a linear function.
Analyte quantification methods with UV-visible spectrophotometers
Depending on the characteristics of the analysis, different methods may be used:
- Signal-to-noise ratio method: it is mainly performed using the UV-visible spectrophotometer. It requires the procedure to provide a white signal, background noise or baseline, i.e. a residual signal at zero analyte concentration.
- Method of standard deviation of the blank response and the slope of the calibration line: the detection limit and the limit of quantification of the method can be calculated by knowing the standard deviation attributed to the sample response and the slope of the calibration line of the analyte. The mathematical expression to adjust the signal is done depending on the equipment, for this case the UV-visible spectrophotometer.
- Extrapolation method of the calibration line at zero concentration: using the slope of the calibration line, the actual blank value is replaced and extrapolated from the calibration line.
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