|Year : 2017 | Volume
| Issue : 1 | Page : 21-25
Analysis of antioxidant activity and total phenolic content of some homoeopathic mother tinctures
Saeed Ahmad1, Tayyeba Rehman2, Waheed Mumtaz Abbasi2, Muhammad Mohsin Zaman3
1 Department of Pharmacy, Faculty of Pharmacy and Alternative Medicine, The Islamia University of Bahawalpur, Bahawalpur, Pakistan
2 Department of Pharmacy, Faculty of Pharmacy and Alternative Medicine, University College of Conventional Medicine, The Islamia University of Bahawalpur, Bahawalpur, Pakistan
3 Department of Biochemistry, Bahauddin Zakariya University, Multan, Punjab, Pakistan
|Date of Web Publication||23-Feb-2017|
Department of Pharmacy, Faculty of Pharmacy and Alternative Medicine, The Islamia University of Bahawalpur, Bahawalpur
Source of Support: None, Conflict of Interest: None
Background: Antioxidant compounds are widely used in health protection. Homoeopathic mother tinctures are commonly prescribed for prevention and cure of many illnesses. Objective: The present study focused to determine the antioxidant potential of six commonly prescribed mother tinctures. i.e. Syzygium jambolanum, Damiana, Cinchona officinalis, Chelidonium majus, Convallaria majalis, Coca. Materials and Methods: Antioxidant activity was estimated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) inhibition assay. A volume of 0.1 mM solution of DPPH was used while mother tinctures (5, 2.5, and 1.25 μl volumes) were used for estimating antioxidant activity. Quercetin was taken as a standard control in antioxidant activity. Total phenolic content was measured by Folin–Ciocalteu reagent assay. Total phenolic content of mother tinctures was measured in comparison to gallic acid. Results: Results of the study showed the significant antioxidant activity and high total phenolic content of all tested mother tinctures in the following order: Syzygium jambolanum, Damiana, Cinchona officinalis, Chelidonium majus, Convallaria majalis, Coca. Conclusion: Antioxidant potential of mother tinctures is related to total phenolic content present in them, and their role in prevention and cure of diseases may link through their antioxidant activity. Among six selected mother tinctures, S. jambolanum has highest antioxidant potential.
Keywords: Syzygium jambolanum, Damiana, Cinchona officinalis, Chelidonium majus, Coca, Convallaria majalis, Turnera diffusa
|How to cite this article:|
Ahmad S, Rehman T, Abbasi WM, Zaman MM. Analysis of antioxidant activity and total phenolic content of some homoeopathic mother tinctures. Indian J Res Homoeopathy 2017;11:21-5
|How to cite this URL:|
Ahmad S, Rehman T, Abbasi WM, Zaman MM. Analysis of antioxidant activity and total phenolic content of some homoeopathic mother tinctures. Indian J Res Homoeopathy [serial online] 2017 [cited 2018 May 27];11:21-5. Available from: http://www.ijrh.org/text.asp?2017/11/1/21/200843
| Introduction|| |
Antioxidant compounds are an important factor in health protection. Moreover, they reduce the risk of chronic diseases such as heart diseases and cancer. Polyphenolic compounds have strong antioxidant activity, whereas monophenols are weak antioxidants. Antioxidants have the ability to trap free radicals. In the biological systems, highly reactive free radicals are present from many sources. These free radicals have the ability to cause degenerative diseases by oxidizing proteins, lipids, nucleic acids, and DNA. Polyphenols, flavonoids, and phenolic acids have the power of scavenging these free radicals and thus helpful in preventing degenerative disorders, aging, etc.
Homoeopathic mother tinctures are prescribed commonly in Homoeopathy  as many mother tinctures can be used as antiseptics. Mother tincture or lower potencies of Convallaria majalis are used in heart failure. It is used as a heart tonic and increases the contractile power of heart.Syzygium jambolanum mother tincture is useful against diabetes mellitus. It helps in managing the high blood sugar. Some studies also showed ameliorating effects of S. jambolanum Q in controlling streptozotocin-induced diabetes in rats.,Coca is described in Homoeopathy as a useful remedy for aging and imbecility.,Chelidonium majus is a prominent liver remedy in Homoeopathy and is useful for various diseases of this organ.Cinchona officinalis is beneficial for debility from exhaustive discharges of any kind. Tincture of Damiana is clinically indicated in sexual debility that is originated from nervous prostration and impotency.
The aim of the study is to estimate antioxidant activity and total phenolic content of six commonly used mother tinctures that is Syzygium jambolanum, Damiana, Cinchona officinalis, Chelidonium majus, Convallaria majalis, Coca.
| Materials and Methods|| |
Chemicals and medicines procurement
All the homoeopathic mother tinctures were purchased from Dr. Willmar Schwabe GmbH and Co., KG, Germany, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, Folin–Ciocalteu reagent (FCR), gallic acid, and quercetin were purchased from Sigma-Aldrich, Germany.
BioTek ® USA ELISA microplate reader, 96-well plates (Pyrex, Japan), and micropipette (HTL, Japan).
2,2-Diphenyl-1-picrylhydrazyl free radical scavenging assay
The scavenging activity of homoeopathic mother tinctures was checked with DPPH radical. A volume of 0.1 mM solution of DPPH was prepared by adding 25 mg of DPPH in 100 ml of methanol. A microassay of antioxidant determination was used with some modifications. A volume of 90 μl of DPPH solution and different serial dilutions (5, 2.5, and 1.25 μl) of mother tincture were placed in wells of a 96-well microplate. The wells containing DPPH and 10 μl of ethanol were used as negative control. Quercetin was taken as standard control. The whole process was done in triplicate. The reaction mixture was placed for 30 min at 37°C. The absorbance was measured at 517 nm using Biotech ELISA microplate reader. The absorbance was taken as follows:
Scavenging activity % = ([Ac − Am.t)/Ac] × 100
Ac = Absorbance of negative control
Am.t = Absorbance of mother tincture
Total phenolic content estimation
Total phenolic content was estimated by FCR (Sigma-Aldrich, Germany) using the methodology of McDonald. A volume of 5 µl of mother tinctures and 50 µl of 1 mM sodium carbonate were added and then Folin–Ciocalteu 20% was added to make the volume up to 150 µl in 96-well plate, and after incubating at 40°C, the absorbance was checked at 765 nm.
The results were analyzed by Statistical Package of Social Sciences software (IBMSPSSV, 20) Armonk, NY: IBM Corp. One-way analysis of variance followed by post hoc test was applied for checking statistical significance of results. Pearson correlation coefficient test was used for checking any correlation between antioxidant activity by DPPH assay and total phenolic content of mother tinctures. Significance level was set at P ≤ 0.05.
| Results and Discussion|| |
In the present study, all the six mother tinctures were able to decolorize DPPH, and the free radical scavenging potentials of the mother tinctures were found to be in the order of S. jambolanum < Damiana < C. officinalis < C. majus < C. majalis < Coca [Table 1]. S. jambolanum inhibits DPPH free radical, and the higher volume (5 µl) showed 87.2% inhibition and the lowest used volume (1.25 µl) showed 39.9% inhibition of DPPH. Similarly, all other mother tinctures showed volume-dependent inhibition of DPPH. Damiana, C. officinalis, C. majus, C. majalis, and Coca inhibited DPPH 81.6, 69, 68.5, 65.1, and 61, respectively, at 5 µl volume, and this percent inhibition was decreased with the decrease of volume and mentioned in detail in [Table 1].
|Table 1: Results of mother tinctures, quercetin, and alcohol against 2,2-diphenyl-1-picrylhydrazyl|
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Different assays can be used to assess antioxidant activity. However, the use of free radical DPPH is the preferred method to measure antioxidant activity as it is a simple and rapid method. Moreover, it is not specific for particular antioxidant component, thus measure the total antioxidant capacity of sample. Hence, antioxidant activity of these mother tinctures is the estimate of their total antioxidant capacity.
DPPH free radical contains an odd electron and is purple in color. The color changes from purple to yellow when the odd electron of DPPH radical combined with hydrogen from a free radical scavenging antioxidant to form the reduced DPPH-H.
The total phenolic contents of the mother tinctures are determined by Folin–Ciocalteu method are reported as gallic acid equivalents [Table 2]. Among the six mother tinctures, S. jambolanum was containing the highest (94.5% ±3.3%) amount of phenolic compounds, followed by Damiana (77.3% ±3.3%), C. officinalis (75.5% ±4.0%), C. majus (45.0% ±5.0%), C. majalis (39.8% ±2.4%), and Coca (37.4% ±2.9%).
The content of total phenolic was carried out based on the absorbance values of the various mother tinctures, reacted with Folin–Ciocalteu reagent, and compared with the standard solutions of gallic equivalents. The way of measuring total phenolic content is based on chemical reducing capacity of a tested sample relative to gallic acid. It is not the absolute measurement of total phenols in a sample.
There is a positive relationship between antioxidant capacity and total phenolic content of the various mother tinctures. In this study, the results showed that mother tinctures contain a significant amount of phenolics, and it is the amount of phenolics present in this extract being responsible for its marked antioxidant activity. Pearson correlation coefficient showed a significant relationship between total phenolic content and DPPH inhibitory assay (P = 0.004) as described in [Table 3] and [Figure 1]. Several reports have shown the close relationship between total phenolic content and antioxidant activity of the fruits, plants, and vegetables. Many phenolic derivatives' compounds are the vital antioxidants which exhibit scavenging efficacy on the free radicals and reactive oxygen species and commonly distributed in the plant kingdom. Hence, antioxidant activity of these mother tinctures may be due to their high phenolic content. Antioxidants are used to prevent aging, diabetes, heart diseases, cancer, and many other illnesses; strong potential of tested mother tinctures as antioxidants in the present study suggests that effect of mother tinctures in various diseases' treatment may be due to their antioxidant activity.
|Table 3: Correlation between phenolic content and 2,2.diphenyl.1.picrylhydrazyl inhibition by mother tinctures|
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|Figure 1: Representation of calculated values of total phenolic content and percentage inhibition of 2,2-diphenyl-1-picrylhydrazyl by mother tinctures (5 μl). Pearson correlation was used to estimate the relation between total phenolic content and 2,2-diphenyl-1-picrylhydrazyl inhibition (Sig [2-tailed] 0.004). Correlation was significant at the 0.01 level (2-tailed)|
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| Conclusion|| |
The results of the study showed antioxidant potential of all tested mother tinctures. These mother tinctures are being used in prevention and treatment of different diseases, and their use in clinical practice may have link through their antioxidant activity. The study emboldens the clinical use of these mother tinctures with more guarantee of their efficacy as all the selected mother tinctures showed antioxidant potential. Among six selected mother tinctures, S. jambolanum has highest antioxidant potential. The antioxidant activity is correspondingly related to concentration of total phenolic content, and it is perhaps the presence of phenolic content in mother tinctures that may be responsible for their antioxidant activity.
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[Table 1], [Table 2], [Table 3]