|Year : 2023 | Volume
| Issue : 1 | Page : 18-25
Preparation, standardization, and in vitro anti-inflammatory action of Navatchara aakiranam: A Siddha therapeutic snuff
Pooja Muniraj, Kirutisri Karthigaeyan, Kehren R Rayma, Aaliya Parveen, Balagurusamy Karthigaeyan
Velumailu Siddha Medical College and Hospital, Sriperumbudur, Tamil Nadu, India
|Date of Submission||27-Oct-2022|
|Date of Decision||22-May-2023|
|Date of Acceptance||24-May-2023|
|Date of Web Publication||29-Aug-2023|
2/7, B.R.G. Madhepalli Village & Post, Bargur, Krishnagiri 635 104, Tamil Nadu
Source of Support: None, Conflict of Interest: None
Background: Siddha system is a traditional system belonging to South India. An estimated 134 million Indians are thought to have chronic sinusitis, which can cause incapacitating headaches, fever, and nasal congestion and blockage, among other symptoms. Aakiranam is one among 32 types of external medicine that is therapeutically more effective due to its quicker mode of absorption. However, these traditional formulations are less validated scientifically. Objective: This study aims at the preparation, standardisation and evaluation of in vitro anti-inflammatory activity of Navatchara aakiranam (NA), a Siddha classical Herbomineral formulation. Materials and Methods: Physicochemical analysis, sophisticated instrumental analysis, such as Fourier transform infrared spectroscopy, scanning electron microscopy, energy dispersive x-ray analysis, and inductively coupled plasma- optical emission spectroscopy studies, and in vitro anti-inflammatory using bovine serum albumin study were conducted on NA. Results: The study drug NA selected from the Siddha classical text “Gunapadam - Thathu Jeeva Vagupu” by Dr. Thiyagarajan R.L.I.M is taken for the first preliminary study as it is a potential as snuff in the treatment of sinusitis. Conclusion: This study validates the standardization of NA through organoleptic, biochemical, physicochemical, microbial load, and instrumental analyses.
Keywords: Anti-inflammatory, Navatchara Aakiranam, Siddha, sinusitis, standardization
|How to cite this article:|
Muniraj P, Karthigaeyan K, Rayma KR, Parveen A, Karthigaeyan B. Preparation, standardization, and in vitro anti-inflammatory action of Navatchara aakiranam: A Siddha therapeutic snuff. J Res Siddha Med 2023;6:18-25
|How to cite this URL:|
Muniraj P, Karthigaeyan K, Rayma KR, Parveen A, Karthigaeyan B. Preparation, standardization, and in vitro anti-inflammatory action of Navatchara aakiranam: A Siddha therapeutic snuff. J Res Siddha Med [serial online] 2023 [cited 2023 Oct 3];6:18-25. Available from: http://www.jrsm.in/text.asp?2023/6/1/18/384577
| Introduction|| |
The Siddha system of medicine is a reputable system belonging to South India. This system has a collection of 32 types of external therapeutic measures (Pura Marunthugal) and internal medicines (Ul marunthugal). Though this ancient system has been utilized by Indians and has gained attention worldwide, it also has an erroneous claim that it fails to faster relief from ailments. Aakiraanam is one among the 32 types of external medicine that is therapeutically more effective due to its quicker mode of absorption, but these traditional formulations are less validated scientifically. Moreover, there is a strive for global acceptance due to the lack of scientific validation and documentation; to overcome the limitations and ensure quality, safety, and therapeutic efficacy, modern methods can be incorporated. Normally, raw drugs undergo the process of purification, trituration, incineration, and calcination to get the end product. It is, therefore, high time to standardize the drugs to assess its quality and purity. This first preliminary study focuses on preparing the traditional therapeutic nasal snuff Navatchara Aakiraanam (NA) that has two ingredients Navatcharam (ammonium chloride) and Seemai sunnambu (calcium carbonate). This is a unique preparation because it can be used as both external medicine and internal medicine. This has provoked the curiosity to prepare and standardize this formulation by physicochemical analysis and using sophisticated instruments.
| Materials and methods|| |
Study drug selection
The study drug Navatchara Aakiranam was selected from the Siddha classical text “Gunapadam Thathu Jeeva Vagupu” by Dr. Thiyagarajan R. L. I. M.
Identification and authentication
All raw drugs are identified and authenticated by the experts of Gunapadam at Velumailu Siddha Medical College and Hospital, Sriperumbudur.
Study drug preparation
Raw Navatcharam was dissolved in cow’s urine and filtered by using a filter. The dissolved Navatcharam is poured in a clay pot, boiled until fluid evaporates, and then exposed to sunlight to dry. Quantity of drugs used are mentioned in the [Table 1].
Navatcharam and Seemai Sunnambu [Figure 1] and [Figure 2] were dried carefully. Both the contents were mixed and sublimated in high fire (kadakini) for 9 h [Figure 3] to obtain the final drug Navatcharam pathangam [Figure 4].
Inhalation via nose can treat headache and sinusitis. It is taken internally to treat nipple fissure, eclampsia, epilepsy, inguinal lymphadenopathy, chicken pox, and respiratory diseases. This medicine is useful for skin diseases such as eczema, herpes, and leprosy.
Rationale behind selecting study drug
Previous studies revealing the individual pharmacological action of preparatory drugs used in NA are listed in [Table 2].
| Results|| |
Herbomineral formulation NA is subjected to various physicochemical parameters for standardization. The results are exhibited in [Table 3].
The loss on drying test is to determine the amount of water and volatile matter in a sample when the sample is dried under the specified conditions. Moisture is one of the major factors responsible for the deterioration of the drugs and formulations. Low moisture is always desirable for higher stability to the drugs. Moisture content of the drug NA is 8.90 ± 0.120%, which depicts the extensive shelf life of the drug.
the total ash value measures the total inorganic content present in the drug. In the drug NA, it has about 91.20% of inorganic content.
The amount of siliceous matter present in the sample is calculated by the amount of acid-insoluble ash value. The lower the acid-insoluble value, the higher the quality of the drug. The acid-insoluble ash of this drug is presented to be 6.10 ± 0.020%.
The water-soluble extractive value of NA is 8.90 ± 0.510 gives the percentage of soluble matters present in the drug. Suitable solvent could be selected based on the extractive value. In addition, it gives the percentage of drug that will interact with the metabolism reactions.
Fourier transform infrared spectroscopy
The results of Fourier transform infrared spectroscopy-NA are discussed in graph [Figure 5]. The absorbance peaks around 3415 cm−1 correspond to the O-H of water/alcohol/carboxylic acid. The absorbance frequency around 2921 and 1787 cm−1 arises from the -C-H and C-H stretches. Furthermore, the bands around 2521 and 712 cm−1 correspond to the bending frequency of C-H stretch, CH2 Bend, C-OH, and C-Cl stretch.
|Figure 5: Graphical representation of Fourier transform infrared spectroscopy |
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Scanning electron microscope
The Scanning electron microscope (SEM) images that were obtained from various regions of the sample N1 clearly depict that the size was commonly found to be ranging between 71.70 and 180.6 µm at 5K magnification [Figure 6] and [Figure 7].
|Figure 6: Scanning electron microscope image of raw Navatcharam (N1)—cluster view and categorized view|
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The SEM images [Figure 8][Figure 9][Figure 10] that were obtained from various regions of the sample N2 clearly depict that the size was commonly found to be ranging between 258 and 390 nm at 5K magnification.
|Figure 8: Scanning electron microscope image of Navatchara aakiranam (N2)|
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|Figure 9: Scanning electron microscope image of Navatchara aakiranam (N2)—cluster view with size|
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|Figure 10: Scanning electron microscope image of Navatchara aakiranam (N2)—categorized view with size|
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Saint Thirumoolar’s hypothesis about atomic theory has been reinstated as nanotechnology in this decade. Nanoscience and technology may be new to the world, but Indian system of medicine has been using this technology to make Parpam, Chendhooram, etc, from time immemorial. Our medicine particles are in oxygen-deficient state, and clearly identifiable fractions of the particles are in the nanometer size range. These properties might impart the therapeutic property.
“மேவிய சீவன் வடிவது சொல்லிடில்
கோவின் மயிரொன்று நூறுடன் கூறிட்டு
மேவிய கூறது ஆயிர மாயினல்
ஆவியின் கூறு நூராயிரத் தொன்றாமே”
Modern medicine cures a particular disease by targeting exactly the affected zone and transporting the drug to that area known as drug delivery system (DDS). By DDS, we get to know about pharmacokinetics, pharmacodynamics, nonspecific toxicity, and immunogenicity of the drug and evaluation of efficacy of drugs and make biorecognition. This type of DDS is a major lacuna in our system. To overcome this, we approach our drug as novel DDS (NDDS).
From the above references, we come to know that our formulations are in nanoparticle size, and they can easily fulfill the NDDS. Through the administration, our drugs mechanisms can be with passive/ active target and releases controlled, potential and pulsatile manner and also as protein – peptide/ lipid – polymer based DDS.
Energy Dispersive X-Ray Analysis
The energy dispersive x-ray analysis (EDAX) results showed the presence of the following elements, as shown in [Table 4] and [Figure 11] and [Figure 12].
Inductively coupled plasma-optical emission spectroscopy
The sample NA drug of 0.5 g (wt: 0.450010 g) is measured and then dissolved in a decomposition vessel with 10-mL of nitric acid solution. The intensity of emission is indicative of the concentration of the element within the sample. Partial spectral profile and analysis results are shown above [Table 5].
|Table 5: Partial spectral profile and analysis results of inductively coupled plasma- optical emission spectroscopy|
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From the above results, the heavy metals, such as arsenic, lead, and mercury, are in below detectable limit. Furthermore, the drug contains calcium, potassium, sulfur, chloride, and phosphorus. It is inferred that all the toxic heavy metals are removed, ensuring the safety of the drug NA. With this, the Siddhar’s admirable formulas are once again proved that the toxic minerals were converted into nontoxic medicines.
In vitro anti-inflammatory assay
The maximum protein [bovine serum albumin (BSA)] denaturation inhibition was 54.6% at 80 µg/mL concentration [Figure 13]. The IC50 values were calculated as 73.91 µg/mL concentration for protein (BSA) to inhibit the denaturation. Protein denaturation, a process in which proteins lose their tertiary structure and secondary structure, is a well-documented cause of inflammation. When denatured, most biological proteins lose their biological functions. As part of the investigation on the anti-inflammatory activity, ability of this drug NA inhibits inflammation at very a low concentration. Results are presented above [Table 6].
|Table 6: Percentage of antidenaturation activity of Navatchara aakiranam|
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Standardization of NA includes physical characterization, such as pH, total ash, water-soluble ash, acid-insoluble ash, and loss on drying, that ensures the drug safety.
Fourier transform infrared spectroscopy shows the presence of functional group through their stretch and bends, which are responsible for its functional activities. The absorbed frequency and the respective probability functional groups are represented above. The main functional groups present are amide, phenols and alcohols, alkanes, aldehyde, amine, alkenes, alkanes, ester, ether, and alkyne.
In inductively coupled plasma-optical emission spectroscopy, safety of this drug is ensured by analysis of heavy metals that are in permissible limits.
In SEM report, the particle N2 showed the reduction in size than N1. Nanoparticles present in the drug are better in its absorption and pharmaceutical action. EDAX showed the content of minerals such as Cl, N, O, C, and Ca, which may be responsible for pharmacological action.
NA has significant anti-inflammatory activity at the dosage of 73.91 µg/mL concentration, and the drug is safe to use. These results could be a better reference for setting limits for the quality assessment of the drug as well as enhancing the therapeutic efficiency in future.
| Conclusion|| |
Through this recent research reports, the efficacy of the medicine NA has ensured stability and genuinity in standardization. Hence, the standardization of the drug is the first step for further assessing toxicological and validating pharmacological activities. NA should be subjected to further preclinical and clinical studies to validate its efficacy and safety certifying the widespread acceptance among public and scientific communities to use the drug for many diseases. Therefore, the drug can be taken to the next level of isolation of the active principles, which is responsible for the therapeutic effect.
The authors would like to sincerely thank the Management, Principal, Vice principal, and Guide of Velumailu Siddha Medical College and Hospital, Sriperumbudur.
Financial support and sponsorship
This study was financially supported by The Ministry of AYUSH, CCRS.
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11], [Figure 12], [Figure 13]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]