Research Article

Physicochemical Studies of Homoeopathic Formulations of Ammonium Aceticum by using Volumetric, Acoustic and Viscometric Measurements at Different Temperatures

Anil Kumar Nain¹*, Neha Chaudhary¹, Anil Khurana², Raj Kumar Manchanda² and Debadatta Nayak²

¹Department of Chemistry, Dyal Singh College, University of Delhi, India

²Central Council for Research in Homeopathy, Ministry of AYUSH, India

Submission: February 13, 2023;Published: March 01, 2023

*Corresponding author: Anil Kumar Nain, Department of Chemistry, Dyal Singh College, University of Delhi, New Delhi – 110 003, India

How to cite this article: Anil Kumar Nain, Neha Chaudhary, Anil Khurana, Raj Kumar Manchanda and Debadatta Nayak. Physicochemical Studies of Homoeopathic Formulations of Ammonium Aceticum by using Volumetric, Acoustic and Viscometric Measurements at Different Temperatures. Organic & Medicinal Chem IJ. 2023; 12(1): 555830. DOI: 10.19080/OMCIJ.2023.12.555830

Abstract

Ocean covers about one-third of the earth and is considered as a vital natural resource. Seaweeds accompany a large areas of the ocean coastal area, which are considered as among the primary producers. Many seaweeds have food values and so many indigenous populations relay on them as their nutrient source. Besides, seaweeds are also rich in many bioactive compounds having pharmacological significance. Seaweeds contain higher number of proteins, vitamins, minerals, essential fatty acids, polysaccharides and so on. Gracilariopsis lemaniformis is a red sea alga and is also rich in bioactive compounds having medicinal values. In this review article, we focused on the pharmacological importance of this species based on recent updates of literature survey and we discussed here about its antimicrobial, antitumor, antidiabetic and antioxidant properties, which would demand its therapeutic relevance for future drug development.

Keywords: Covid-19; Sars-Cov-2; Copper Metal; Cancer; Viral Infections

Introduction

Homoeopathy is one of the traditional and most widespread alternative systems of medicine after conventional therapy. The efficacy of homoeopathic medicines is well supported by research evidence; however, there are controversies regarding implausibility in biological activity of homoeopathic medicines in which the source drug is diluted beyond Avagadro’s limit, i.e., the highly diluted medicine formulation must be identical to the solvent. There have been a few research studies pertaining to explore the presence of drug in extremely diluted formulations [1-7] and its mechanism of action, but the question still remains answered acceptably. The measurements of physicochemical properties and derived parameters of aqueous and mixed-aqueous solutions of amino acids, carbohydrates, drugs, etc. have been supportive in characterizing the solute-solute and solute-solvent interactions, which are subsequently useful in understanding of solute-solvation/hydration behavior of solute and preferential solvation of solute by the solvent [8-15]. Since homoeopathic formulations are extremely dilute solutions, their physicochemical properties, like density, ultrasonic speed, viscosity, refractive index, etc. can be measured easily as a function of concentration/potency and temperature.

The physicochemical properties calculated from these experimental data can deliver valuable information on the understanding of physicochemical behavior and mechanism of drug action. Recently, there have been few physicochemical studies on extremely diluted solutions of inorganic salts [16-21] and homoeopathic medicines [2,3,22-28] by using physicochemical methods. These studies provided interesting and convincing information on the behavior of these extremely diluted solutions.

To the best of our information, very few physicochemical studies on homoeopathic medicines using volumetric, acoustic, viscometric and optical methods have been reported in the literature [2,3,22]. These considerations led us to undertake the present study on the physicochemical behavior of extremely diluted homoeopathic formulations of ammonium aceticum. Ammonium aceticum dilution is helpful in the treatment of many types of conditions, viz., heavy feeling of the head and scraping sensation of the throat, stomach and reducing sweat. It also helps in reducing urination urge.

The homoeopathic medicine formulations are generally obtained through the combination of two processes: a dilution of 1:100 in mass followed by succussion. In the present study, the densities, ρ, ultrasonic speeds, u and viscosities, η of pure ethanol control (91% ethanol in water) and 33 formulations of ammonium aceticum with potencies ranging from 1C to 200C (with intervals of 2C till 30C, and then with intervals of 10C till 200C) at 293.15, 298.15, 303.15, 308.15, 313.15 and 318.15 K and atmospheric pressure. From these experimental data, the isentropic compressibilities κ_s, intermolecular free length, L_f , acoustic impedance, Z, relative association, RA, deviations in isentropic compressibility, deviations in intermolecular free length, deviations in acoustic impedance, and deviations in viscosity, have been calculated. The results have been qualitatively discussed in terms of interactions/physicochemical behavior of ammonium aceticum in these homoeopathic formulations.

Experimental

Ethanol control and homoeopathic formulations of various potencies of ammonium aceticum used in the study were procured from Dr. Wilmer Schwabe India Pvt. Limited, India. The densities and ultrasonic speeds of the samples were measured by using high precision digital vibrating tube Density and Sound Analyzer (DSA 5000M, Anton Paar, Austria). This two-in-one instrument is equipped with both density and ultrasonic cells, with reproducibility of ±1 × 10⁻³ kg·m⁻³ and ±1 × 10⁻² m·s⁻¹ for density and ultrasonic speed, respectively. The temperature for both cells was kept constant by using built in peltier thermostat within ±0.01 K. The calibration of instrument was done by using triply-distilled, degassed water and with dry air at atmospheric pressure and 293.15 K [11]. The operating working frequency used for ultrasonic speed measurements is 3 MHz. The principle used in density measurement is based upon oscillating U-tube principle while the speed of sound is measured using a propagation time technique. The standard uncertainties related to the measurements of density, ultrasonic speed and temperature were found within ±0.05 kg·m⁻³, ±0.5 m·s⁻¹ and ±0.01 K, respectively.

The viscosity measurements were done by using microviscometer (Lovis 2000M, Anton Paar, Austria) at temperatures, (293.15 − 318.15) K, and atmospheric pressure p = 101 kPa. The temperature was controlled to ±0.02 K by an automatic build in Peltier technique . The rolling ball principle was used in the measurement of viscosity, having a calibrated glass capillary with a steel ball as supplied by manufacturer. The calibration of capillary was accomplished by using viscosity standard fluids. The relative standard uncertainty in viscosity measurements was found to be within ±0.5 %.

Results

The experimental values of densities, ρ, ultrasonic speeds, u and viscosities, η of ammonium aceticum formulations as function of potency (in centesimal) at different temperatures are listed in Tables 1-3, and are presented graphically in Figures 1-3, respectively. The values of the isentropic compressibility, intermolecular free length, acoustic impedance, Z and relative association, R_A have been calculated by using the following relations

where K’ is temperature dependent constant [= (93.875 + 0.375T) × 10−8]; T is the absolute temperature, ρo, ρ, uo and u are the densities and ultrasonic speeds of the solvent and the solution, respectively. The values of κ_s, L_f , Z and RA are given in Tables 4-7. The deviations in κ_s, L_f , Z, η and n_D of ethanol due to addition of ammonium aceticum with dilution and succussion are represented by deviation values of these properties. The deviations in isentropic compressibility, Δκ_s , deviations in intermolecular free length, ΔL_f , deviations in acoustic impedance, ΔZ and deviations in viscosity, Δη have been calculated by using the following standard relations

where the superscript ‘o’ represents the values for pure ethanol control (91% ethanol in water). The variations of Δκ_s , ΔL_f , ΔZ and Δη with potency, C of ammonium aceticum and temperature are presented graphically in Figures 4-7, respectively.

Discussion

A close perusal of Tables 1-3 and Figures 1-3 indicates that the values of ρ and u and η of ammonium aceticum in ethanol are more than those of ethanol control for all the potencies (1C to 200C) at each investigated temperature and the values decrease with increase in temperature. The values of ρ and u are maximum at 1C and then decrease significantly in presence of ammonium aceticum for simple successive dilution to the potency 4C and after that these values increase slightly till 8C and remain nearly constant up to potency 22C and then decrease and exhibit a minimum at 26C and thereafter 30C the values remain nearly constant up to 180C, and thereafter, again increase significantly after potency 180C up to 200C (Figures 1 & 2). The values of values of η are maximum at 1C decrease significantly in presence of ammonium aceticum for simple successive dilutions from potency 1C to 4C and then increase up to 8C and thereafter remain nearly constant up to potency 22C and then decrease and exhibit a minimum at 26C and then after 30C the values remain nearly constant up to 180C. Thereafter, values of η again increase after potency 180C up to 200C (Figure 3). The observed anomalous trends in ρ, u and η at certain potencies, viz., 1C to 4C, 22C to 30C and 180C to 200C indicate that these potencies exhibit different solution structure as compared to other potencies and ethanol control.

A close perusal of Tables 4 & 5 indicate that the values of κ_s and L_f for ammonium aceticum potencies are less than those of ethanol controls for all the potencies at each investigated temperature and these values increase with increase in temperature, which indicates significant interaction between ammonium aceticum and ethanol molecules. The values of κ_s and L_f decrease significantly in presence of ammonium aceticum for simple successive dilutions up to potency 4C and after that value pass through maxima/minima at some potencies with dilution and these values again decrease after potency 180C up to 200C (Tables 4 & 5). These variations in κ_s and L_f are expressed in terms of deviations in isentropic compressibility, Δκ_s and deviations in intermolecular free length, ΔL_f and are shown graphically in Figures 4 & 5.

Figures 4 & 5 indicate that the values of Δκ_s and ΔLf are negative and these values are minimum for potency 1C and then increase till potency 4C and after that values decrease till potency 8C and then remain nearly constant on more dilution till 22C, thereafter these values increase and exhibit a maximum at 26C and then after 30C the values remain nearly constant up to 180C, finally decrease again after potency 180C up to 200C. This indicates that at potencies 2C to 6C, 24C to 28C are more compressible and 1C, 190C to 200C are less compressible than the other potencies, indicating that the potencies 2C to 6C, 24C to 28C exhibit less compact solution structure and the potencies 1C, 190C to 200C exhibit less compact solution structure as compared to other potencies and ethanol control. The minimum in Δκ_s and ΔL_f values at potencies 1C, 190C and 200C indicate that these have most compact solution structure as compared to other potencies, hence, these potencies may have different behavior in terms of efficacy when used in practice.

A close perusal of Table 6 and Figure 6 indicate that the acoustic impedances, Z of potencies of ammonium aceticum are more than those of ethanol control for all the potencies at each investigated temperature and the values decrease with increase in temperature, which indicates significant interaction between ammonium aceticum and ethanol molecules. These variations in Z are expressed in terms of deviations in acoustic impedance, ΔZ and are shown in Figure 6. it indicates that ΔZ values are positive, i.e., Z values for ammonium aceticum are more than those of ethanol control. These ΔZ values are maximum for potency 1C and then decrease till potency 4C and after that values increase till potency 8C and then remain nearly constant on more dilution till 22C, thereafter these values decrease and exhibit a minimum at 26C and then after from 30C the values remain nearly constant up to 180C, finally increase again after potency 180C up to 200C.

This indicates that at potencies 2C to 6C, 24C to 28C offer less impedance to sound waves due to less compact structure and 1C, 190C to 200C offer more impedance to sound waves due to more compact structure than the other potencies and ethanol control. The variations in values of Z and ΔZ of these potencies may be due interaction between ammonium aceticum and ethanol molecules.

A close perusal of Tables 1-3 and Figures 1-3 indicates that the values of ρ and u and η of ammonium aceticum in ethanol are more than those of ethanol control for all the potencies (1C to 200C) at each investigated temperature and the values decrease with increase in temperature. The values of ρ and u are maximum at 1C and then decrease significantly in presence of ammonium aceticum for simple successive dilution to the potency 4C and after that these values increase slightly till 8C and remain nearly constant up to potency 22C and then decrease and exhibit a minimum at 26C and thereafter 30C the values remain nearly constant up to 180C, and thereafter, again increase significantly after potency 180C up to 200C (Figures 1 & 2). The values of values of η are maximum at 1C decrease significantly in presence of ammonium aceticum for simple successive dilutions from potency 1C to 4C and then increase up to 8C and thereafter remain nearly constant up to potency 22C and then decrease and exhibit a minimum at 26C and then after 30C the values remain nearly constant up to 180C. Thereafter, values of η again increase after potency 180C up to 200C (Figure 3). The observed anomalous trends in ρ, u and η at certain potencies, viz., 1C to 4C, 22C to 30C and 180C to 200C indicate that these potencies exhibit different solution structure as compared to other potencies and ethanol control.

A close perusal of Table 7 and Figure 7 indicate that the viscosities, η of potencies of ammonium aceticum are more than those of ethanol control for all the potencies at each investigated temperature and the values decrease with increase in temperature, which indicates substantial interaction between ammonium aceticum and ethanol molecules. These variations in η are expressed in terms of deviations in acoustic impedance, Δη and are shown in Figure 7. it indicates that Δη values are positive, i.e., η values for ammonium aceticum are more than those of ethanol control. These Δη values are maximum for potency 1C and then decrease till potency 4C and after that values increase till potency 8C and then remain nearly constant on more dilution till 22C, thereafter these values decrease and exhibit a minimum at 26C and thereafter from 30C the values remain nearly constant up to 180C, finally increase again after potency 180C up to 200C. The variations in values of η and Δη of these potencies may be due interaction between ammonium aceticum and ethanol molecules. It is observed that the variations observed in the values of measured properties, ρ and u and η ; and calculated parameters, κ_s, L_f , Δκ_s , ΔL_f , ΔZ and Δη support each other.

A close perusal of Table 7 indicates that the values of RA for 1C, 190C and 200C potencies of ammonium aceticum are more than 1 while for all other potencies the values are less than 1, and other potencies show varying values with slight variations at certain potencies. The changes in values of RA of solution in presence of ammonium aceticum are due to different extents of breaking/ formation of hydrogen-bonded associates in ethanol controls and their interaction with ammonium aceticum with successive dilutions and succussion.

Conclusion

The densities, ultrasonic speeds and viscosities of ethanol control, 33 formulations of ammonium aceticum in ethanol control are measured for potencies from 1C to 200C (with an interval of 2C up to 30C and then of 10C up to 200C) at six different temperatures and atmospheric pressure. From these experimental data, various physicochemical parameters, viz., κ_s ,L_f , Z , Δκ_s , ΔL_f , ΔZ , Δη and RA have been calculated. The results have been qualitatively discussed in terms of interactions/ physicochemical behavior of these extremely dilute homoeopathic formulations of ammonium aceticum in ethanol. The potencies 2C to 6C, 24C to 28C exhibit less compact solution structure and the potencies 1C, 190C to 200C exhibit less compact solution structure as compared to other potencies and ethanol control. Hence, these potencies may have different behavior in terms of efficacy when used in practice. It can be qualitatively concluded that even in extreme dilutions the molecules of ammonium aceticum may be present in these homoeopathic formulations, however it needs to be confirmed from other techniques.

Acknowledgement

The authors are thankful to Central Council for Research in Homeopathy (CCRH), Ministry of AYUSH, Govt. of India for providing financial assistance to carry out this collaborative study.

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