ABSTRACT
OBJECTIVE: This study focuses on standardizing the preparation of homeopathic mother tinctures from the dried bark of four plants—Cinchona officinalis, Holarrhena antidysentrica, Terminalia arjuna, and Viburnum opulus—by evaluating their physicochemical properties and moisture content.
MATERIAL AND METHOD: The tinctures were prepared at the Government Homoeopathic Medical College and Hospital Bhopal and analysed for parameters including alcohol content, pH, specific gravity, weight per millilitres, total solids, HPTLC profiles, and lambda max. The moisture content, an essential factor influencing tincture stability and potency, was also assessed and found to be critical at a threshold of 10%
RESULT: The study showed that the self-prepared tinctures aligned with the established standards in most parameters, with no significant differences in alcohol content, pH, specific gravity, and other measurements compared to the Homoeopathic Pharmacopoeia of India (HPI) standards. However, HPTLC analysis revealed novel chromatographic peaks in Viburnum opulus, suggesting the presence of secondary metabolites not previously documented.
CONCLUSION: The research concludes that while the self-prepared tinctures generally meet HPI standards. The study also emphasizes the need to monitor moisture content to ensure stability and efficacy, suggesting its potential inclusion in future HPI guidelines. Additionally, the discovery of unique chromatographic peaks in Viburnum opulus calls for further investigation into its role in homeopathic treatments.
Keywords: Mother tincture, Cinchona officinalis, Holarrhena antidysenterica, Terminalia arjuna, and Viburnum opulus, moisture content, physiochemical, HPTLC, Lambda max.
INTRODUCTION:
Drug standardization in homoeopathy involves assessing the physio-chemical and pharmacological properties of drugs to ensure quality. In India, the quality of homeopathic drugs is regulated by the Drug and Cosmetics Act, 1940. The Homoeopathic Pharmacopoeia Laboratory (HPL) is responsible for standardizing medicines and publishing the Homoeopathic Pharmacopoeia of India (HPI).1,2,3,4
Moisture Content:
Moisture content in crude drugs is vital for their effectiveness. Excess moisture can cause microbial
spoilage and degradation, while insufficient moisture can reduce potency. Maintaining the right moisture balance is essential for consistent drug strength, as it influences the preparation of tinctures and other homeopathic forms. There is limited research on the impact of moisture content in mother tinctures prepared from dried bark, particularly following HPI standards. Research is needed to understand how drying, storage, and regional factors affect moisture content and its influence on the stability and efficacy of these tinctures.5,6,7,8
Sources of Homeopathic Medicines:
Homeopathic remedies are derived from plants, minerals, animals, nosodes, sarcodes, imponderabilia, and synthetic substances. Plant bark used, with the best quality bark being collected at specific times.9,10
Vehicle:
Absolute alcohol (99.4% ethanol) is the solvent used in preparing tinctures.11
MATERIAL AND METHOD: The present study was conducted at the GHMC Pharmacy Laboratory and RGPV Bhopal, where Thin Layer Chromatography (TLC) and lambda max were performed. Mother tinctures were prepared from the dried bark, which was procured from FSSAI-registered herbal shops as well as wild collections. The study followed an exploratory experimental and comparative approach, analysing four different mother tinctures against the standards prescribed in the Homeopathic Pharmacopoeia of India (HPI) using the maceration method. Various physicochemical parameters, including pH, alcohol content, total solids, High-Performance Thin Layer Chromatography (HPTLC), weight per mL, specific gravity, moisture content, and lambda max, were evaluated.12-29
OBSERVATION
Cinchona officinalis
Macroscopic Characteristics:
The stem bark occurs in quilled or curved pieces, up to 30 cm long and 2–6 mm thick. The outer surface is dull brown-grey or grey, often with lichens and mosses, and marked with transverse fissures. The surface may be furrowed, wrinkled, or fissured, with exfoliation in some varieties. The inner surface is striated, ranging in color from pale-yellowish brown to deep reddish-brown. The fracture is short in the outer layers and fibrous in the inner layers.
Microscopic Characteristics:
The bark contains thin-walled cork cells, numerous isolated yellowish spindle-shaped phloem fibers (up to 90 µm in diameter) with funnel-shaped pits, and a few starch grains (6–10 µm in diameter) from the parenchyma. Parenchymatous idioblasts contain microcrystals of calcium oxalate, while stone cells are rare. The key alkaloid present is quinine. 30
Holarrhena antidyssentrica
Macroscopic Characteristics:
The bark appears as flat or slightly curved pieces, up to 15 cm or more in length, 10 cm in width, and 0.3 to 1 cm thick. The outer surface is grey and smooth, while the inner surface is light grey and finely striated. The fracture is short, revealing a pinkish bark.
Microscopic Characteristics:
The bark consists of thick-walled, radially arranged cork cells filled with tannin. The deep phloem is traversed by uniseriate ray cells and consists of bast fibers, crystal fibers, sieve tubes with companion cells, and phloem parenchyma. Bast fibers occur in groups, surrounded by one or two layers of concentric crystal fibers. Calcium oxalate crystals are mostly spheroidal, and tannin is present in all bark tissues. 31
Terminalia arjuna
The bark occurs in small, recurved pieces with a buff to brownish, wrinkled outer surface and a rough, scaly inner surface. It has a short, granular fracture. Microscopically, it contains cork cells, stone cells with calcium oxalate crystals, non-lignified pericyclic fibers, and a wide secondary phloem with sieve tubes and medullary rays. Found throughout India, it is documented in Drugs of Hindoostan by Dr. S. C. Ghose and was proved at the Midnapore Drug Proving Research Centre. The dried bark is used for preparing the mother tincture.32
Viburnum opulus
acroscopic Characteristics: The drug appears as channeled or quilled pieces, 1–3 cm wide and up to 4 mm thick. The young bark is brownish and smooth with whitish lenticels, while the old bark is brownish-grey, fissured, and scaly. The inner surface is yellowish to reddish-brown with striations or reticulations. Glistening points are visible externally. The fracture is short and granular, revealing a narrow brown bark, a whitish cortex, and yellowish sclerenchyma groups. It has a slight odor and a bitter, astringent taste.
Microscopic Characteristics: Key features include numerous stone cells in ovoid masses, abundant cluster crystals of calcium oxalate in the cortex and phloem, patches of lignified cork cells, absence of phloem fibers, and starch grains up to 15 µm in diameter.33
The barks used in this study are as follows
Cinchona bark Holarrhena bark
Terminalia arjuna bark Viburnum opulus bark
PHYSIOCHEMICAL STUDIES
TABLE1: STANDARDIZATION OF RAW MATERIAL
| Drug substance | Cinchona officinalis | Holarrhena antidyssentrica | Terminalia arjuna | Viburnum opulus |
| Ash | .93% | .96% | .93% | .97% |
| water soluble ash | .87% | .93% | .12% | .97% |
| Sulphated ash | .94% | .91% | .97% |
TABLE 2: FORMULATION OF MOTHER TINCTURE
| s.no. | Name of drug | Cinchona officinalis | Holarrhena antidysentrica | Terminalia arjuna | Viburnum opulus |
| 1 | PART USED | DRIED BARK | DRIED BARK | DRIED BARK | DRIED BARK |
| 2 | DRUG STRENGTH | 1/10 | 1/10 | 1/10 | 1/10 |
| 3 | PREPARATION | ||||
| moderately coarse powder | 100 g | 100 g | 100 g | 100 g | |
| Purified Water | 200 ml | 500 ml | 160 ml | 400 ml | |
| Strong Alcohol | 824 ml | 600 ml | 875 ml | 635 ml | |
to make one thousand millilitres of the Mother Tincture.
TABLE 3: STANDARDIZATION OF MOTHER TINCTURE
| s.no. | Name of drug | Cinchona officinalis | Holarrhena antidysentrica | Terminalia arjuna | Viburnum opulus | ||||
| 3 | PARAMETERS | H.P.I. | SELF PREPARE D | H.P.I. | SELF PREPARE D | H.P.I. | SELF PREPARE D | H.P.I. | SELF PREPARE D |
| 4 | ALCOHOL CONTENT(V/V) | 75.0- 79.0% | 77% | 54.0- 58.0% | 58% | 77.0- 81.0 | 77% | 57.0- 61.0 | 57% |
| 5 | PH | 4.90- 5.40 | 5.46 | 5.0- 6.0 | 7.41 | 4.2-5.0 | 5.36 | 5.60- 6.20 | 5.46 |
| 6 | SPECIFIC GRAVITY | 0.878- 0.880 | 0.868 | 0.85- 0.870 | 0.915 | 0.890- 0.92 | 0.854 | 0.91- 0.93 | 0.898 |
| 7 | TOTAL SOLIDS | not LESS THA 0.80 | 1.197 | 1.0%( w/v) | 1.197 | 1.0%(w /v) | 1.79 | 0.75% (w/v) | 1.2 |
| 8 | HPTLC | 0.24 | 0.24 | 0.2,0. 32,0.6 5,0.83 ,0.93 | 0.2,0.32,0 .65,0.83,0 .93 | 0.05,0.1 2,0.37,0 .72,0.93 | 0.05,0.12 ,0.37,0.7 2,0.93 | Not menti oned | .02,0.12, 0.24,0.39 ,0.49,0.5 9,0.83 |
| 9 | LAMBDA MAX (nm) | 280- 320 | 280 | 270 | 270 | 270 | 270 | 270- 320 | 320 |
| 10. | WEIGHT PER ML(GM) | 0.878 -0.880 | 0.878 | 0.910- 0.930 | 0.934 | 0.850- 0.870 | 0.876 | 0.890- 0.920 | 0.92 |
| 11. | MOISTURE CONTENT | 0.2% | 2% | 1.2% | 8% | ||||
HPTLC
Lambda max
Hollarhena antidysentrica
2
0.90.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 0.10
CINCHONA OFFICINALIS
Viburnum opulus
0.9
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 DISCUSSION
This study was conducted to understand the process of preparing self-prepared mother tinctures from the bark and their standardization in comparison with the H.P.I. parameters. The research was performed in the Pharmacy Laboratory of Government Homoeopathic Medical College and Hospital, Bhopal, except for hpTLC and lambda max analysis, which were conducted in collaboration with RGPV Research Lab, Bhopal, with permission from the Head of the Department (Homoeopathic Pharmacy) at Government Homoeopathic Medical College and Hospital, Bhopal.
Four self-prepared homoeopathic mother tinctures were prepared from dried bark following H.P.I. guidelines. The study involved tincture preparation, storage, labeling, and quality assessment. An exploratory experimental and comparative study approach was adopted, comparing the drug parameters with H.P.I. standards. Statistical analysis was performed using Student’s paired t-test, and the calculated t-values were compared with t-table values for a degree of freedom (df) 5 at p>0.05 to determine statistical significance.
The study emphasizes the importance of moisture content, a critical parameter affecting stability, efficacy, and safety. While HPI does not specify moisture content for dry bark, a 10% moisture content limit was established based on literature review and expert consultation to prevent degradation and microbial growth.
Additionally, HPTLC analysis of Viburnum opulus identified specific Rf values not recorded in the HPI, potentially indicating previously undocumented secondary metabolites. These findings suggest the need for further research to explore their pharmacological potential.
CONCLUSION
This comparative study demonstrated that self-prepared mother tinctures closely match H.P.I. parameters. These findings highlight the effectiveness of standardized preparation methods.
The study also underscores the importance of moisture content regulation, recommending a limit of less than 10% for improved stability and efficacy. Future HPI guidelines should incorporate this parameter to enhance quality control.
Furthermore, the identification of novel Rf values in Viburnum opulus suggests the presence of potentially significant secondary metabolites. Further research is warranted to explore these findings, which may lead to their inclusion in future HPI revisions, thereby expanding knowledge on the therapeutic applications of Viburnum opulus.
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