|
 
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| ORIGINAL ARTICLE |
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| Year : 2023 | Volume
: 8
| Issue : 3 | Page : 280-292 |
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Pharmaceutical standardization of Amrita Bhallataka Rasayana: A compound polyherbal Ayurvedic formulation
Yagnik D Mundadiya, Swapnil Y Chaudhari, Biswa Jyoti Patgiri
Department of Rasashastra & Bhaishajya Kalpana, Institute of Teaching and Research in Ayurveda, Jamnagar, Gujarat, India
| Date of Submission | 12-Jul-2022 |
| Date of Acceptance | 15-May-2023 |
| Date of Web Publication | 16-Aug-2023 |
Correspondence Address:
Yagnik D Mundadiya
Department of Rasashastra & Bhaishajya Kalpana, Institute of Teaching and Research in Ayurveda, Room No. 74, ITRA Boys Hostel, Jamnagar 361008, Gujarat
India
 Source of Support: None, Conflict of Interest: None
DOI: 10.4103/jdras.jdras_109_22
BACKGROUND: Amrita Bhallataka Rasayana (ABR) is one such Ayurvedic polyherbal compound formulation containing Bhallataka as an integral component. It is mainly indicated for rejuvenating purpose. An interesting fact in the preparation of ABR is the quantity of sugar that is to be added as per our convenience and the form of finished product (Avaleha or Ghrita). Earlier research scholars have kept this formulation as Ghrita (medicated ghee) or Avaleha (medicated semisolid preparation) dosage form as the amount of Ghrita is more in this formulation but Paka is to be done as per Avaleha dosage form. Hence, it becomes difficult to prepare and standardize such formulations. To validate the standard manufacturing process (SMP) and quality standards of Amrita Bhallataka Rasayana. METHODS: A total of three batches of ABR were prepared and subjected to organoleptic, physicochemical, high-performance thin-layer chromatography (HPTLC), and nutritional analysis to generate an analytical profile of ABR. In phytochemical qualitative analysis, the presence of saponins, flavonoids, and tannins was assessed by using appropriate qualitative testing methods. RESULTS: Using pertinent physicochemical criteria in accordance with the Ayurvedic Pharmacopeia of India, an analysis of raw and Shodhita Bhallataka was conducted. The alkaline pH value of ABR may be observed due to protein-tannins complexes between milk and oil of Bhallataka. HPTLC was done to develop a chromatographic pattern of three samples of ABR. The average water-soluble extractive of ABR was 54.14%, whereas alcohol-soluble extractive was 19.93%. CONCLUSION: The method of preparation given in the current study for ABR may be considered standard. These preliminary parameters may be helpful as standards for further studies as they are not mentioned in the Ayurvedic Pharmacopeia of India. This study may provide certain leads toward using different proportions of sugar in the preparation of ABR. Keywords: Amrita Bhallataka Rasayana, Bhallataka, Semecarpus anacardium L., Shodhana, standardization
How to cite this article:
Mundadiya YD, Chaudhari SY, Patgiri BJ. Pharmaceutical standardization of Amrita Bhallataka Rasayana: A compound polyherbal Ayurvedic formulation. J Drug Res Ayurvedic Sci 2023;8:280-92 |
How to cite this URL:
Mundadiya YD, Chaudhari SY, Patgiri BJ. Pharmaceutical standardization of Amrita Bhallataka Rasayana: A compound polyherbal Ayurvedic formulation. J Drug Res Ayurvedic Sci [serial online] 2023 [cited 2023 Sep 23];8:280-92. Available from: http://www.jdrasccras.com/text.asp?2023/8/3/280/383692 |
| Introduction | |  |
The fruit of Bhallataka (Semecarpus anacardium L., Anacardiaceae) is a very potent drug having high therapeutic attributes and has been extensively used in indigenous systems of medicines such as Ayurveda prevalent in the Indian subcontinent. Bhallataka is a drug of choice in the treatment of Vata and Kapha predominant disease conditions.[1] It is widely used as an ingredient in many Ayurvedic formulations and being indicated for many ailments.[2],[3] The fruit of Bhallataka is mentioned as a poisonous herbal material in Drugs and Cosmetics Act (India) 1940.[4] It should be administered internally only after Shodhana (purificatory measures). It is well reported to have anti-inflammatory, antibacterial, antioxidant activity, analgesic activity, anticancer activity, antihelminthic activity, and atherogenic activity as well as the Rasayana (rejuvenating) properties when used in different dosage forms.[5] Many compound formulations such as Brihata Bhallataka Avaleha, Bhallatakadi Churna, Bhallataka Kshara, Bhallataka Ghrita, and Amrita Bhallataka Rasayana (ABR) contain Bhallataka as an ingredient.[2],[3],[6],[7] ABR is one such Ayurvedic polyherbal compound formulation containing Bhallataka as an integral component indicated for Rasayana.[8] Some Ayurvedic scholars have kept this formulation as Ghrita or Avaleha dosage form as the amount of Ghrita is more in this formulation but Paka is to be done as per Avaleha dosage form. The amount of sugar in this composition is one remarkable detail. According to our convenience, the classics advise adding a certain amount of sugar to this formulation. So, it becomes difficult to prepare such formulation. Conversion of formulations into various dosage forms to achieve added benefits keeping intact the therapeutic properties has gained momentum in recent past that has great importance in the market. In addition, standardization and quality control of Ayurvedic medicines are also other major concerns in the current era. Considering this, it is planned to standardize ABR on pharmaceutical, physicochemical, phytochemical, and nutritional evaluation grounds.
| Materials and Methods | | |
Procurement of raw materials
Matured fruits of Semecarpus anacardium L. (PubChem SID:472397559) were procured from a pharmacy attached to the Institute. Sharkara (sugar; PubChem CID:5988), Goghrita (clarified butter; PubChem SID:439578207), and Godugdha (cow’s milk; PubChem SID:347910703) of Amul brand were purchased from the local market of Jamnagar, Gujarat. Raw material were authenticated by Pharmacognocist at the Institute.
Preparation of Amrita Bhallataka Rasayana
The entire procedure is divided into the sections listed below. For three batches, every procedure was performed.
Examination of Prashasta-Aprashasta Bhallataka Phala
Acceptable Bhallataka fruits were chosen as per classical guidelines.[9] Physical impurities such as dust, sand, false fruits, and other herbal material of the tree were manually eliminated from Bhallataka fruits. These fruits were dipped in potable water kept in stainless steel vessel. Care was taken that water does not flow out after dipping the Bhallataka fruits. The vessel was left undisturbed for 30 min. Then, Bhallataka fruits, which floated on the water surface, were purposefully removed and kept separately on butter paper for drying. Those Bhallataka fruits, which settled down at the base of vessel, were also removed, kept on butter paper, and dried for 2 days in sun light. Bhallataka fruits, which sunk in water, were used for Shodhana and preparation of ABR. The results of Ashuddha Bhallataka Phala impurities and Prashasta-Aprashasta Bhallataka Phala are depicted in [Table 1] and [Table 2]. | Table 1: Results obtained after removal of physical impurities from Ashuddha Bhallataka Phala
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 | Table 2: Results obtained after examination of Prashasta-Aprashasta Bhallataka Phala
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Shodhana of Bhallataka fruits
Ayurvedic classics have advocated to process Bhallataka fruits in Ishtika Churna (brick powder).[10] Brick was hammered and sieved (no. 10) to get uniform particle sizes, which facilitate further processing. Brick powder was spread in the enamel tray. The pericarps of the acceptable Bhallataka fruits were removed with a cutter, and fruits were kept in brick powder for 7 days. Then, these fruits of Semecarpus anacardium L. were removed from brick powder and cleaned with hot water for three times. Bhallataka fruits were dried in the sunlight for 2 days. After complete drying, they were stored in an airtight glass container. The same Shodhana procedure was followed for another batch of Bhallataka fruits.
Pilot batches of Amrita Bhallataka Rasayana
As classics have given full authority to the physician for the quantity of sugar to be used in this formulation. So, pilot batches of ABR were prepared with different proportions of Sharkara to develop a suitable dosage form. A total of four pilot batches were prepared to get proper consistency of ABR. As the quantity of sugar is fixed, the main three batches of ABR were prepared as per classical guidelines. Details of pilot batches are given [Table 3]. | Table 3: Results obtained during preparation of pilot batches of Amrita Bhallataka Rasayana
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As no specific quantity of sugar has been mentioned in the verse, the first pilot batch of ABR was made with half proportion of sugar than Kwatha, but it was failed as became hard and granular. In the second batch, ingredients were added with classical order, with Sharkara making for three-fourth of the total, followed by Kwatha; however, due to overheating, the material became granular. In the third batch, Sharkara was used one-fourth of Kwatha. But consistency became loose. In the next pilot batch, Kwatha materials were prepared in equal quantities with Sharkara until appropriate Siddhi Lakshanas were achieved. This batch kept its consistency and was semisolid, with crystallized sugar present [Table 3] and [Table 4]. | Table 4: Observations during preparations of pilot batches of Amrita Bhallataka Rasayana
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Preparation of Bhallataka Kwatha
Shuddha Bhallataka fruits were pounded in an iron mortar and pestle before being ground into a coarse powder in a grinder.[10] The contents were added with eight parts (800 mL) of potable water and left undisturbed overnight. On next morning, the contents were boiled over mild flame maintaining temperature between 90°C and 95°C and reduced to one-fourth (200 mL) of its initial volume with constant stirring. The contents were filtered through a clean single-folded cotton cloth to obtain decoction.[10] The residue that remained in the cloth was weighed and discarded. Specific precautions such as application of coconut oil to exposed area and wearing gloves, masks, and goggles were taken during this process.
Preparation of Amrita Bhallataka Rasayana
The formulation composition of ABR is depicted in [Table 5]. The prescribed quantity of boiled Godugdha and Bhallataka Kwatha was allowed for self-cooling. Then, they were shifted together into another stainless steel vessel and subjected to mild heat over liquefied petroleum gas stove. Goghrita was heated (85°C–95°C) in another stainless steel vessel to make it moisture content free. This preheated Goghrita was immediately added in the vessel containing mixture of Godugdha and decoction of Bhallataka with continuous stirring. Sugar was also added in this mixture, and continuous stirring was done till sugar got completely dissolved. The mixture was filtered through a clean cotton cloth to separate undisclosed material, if any. The filtrate was collected into another sterile vessel. This filtrate prepared by addition of sugar in the specified ratio to the Kwatha and Godugdha was heated for 15 min on the 1st day, and remaining Paka (heating) was done on the next day. Heating was stopped after the appearance of Avaleha Siddhi Lakshanas (chief desired characteristics) such as Darvi Pralepa (sticking to the ladle), Apsumajjanam (sinking in water), Patitastu Na Sheeryate (remain stable), RasaGandha Varnotpatti (attaining pleasant taste, odor, and good color). After self-cooling, it was kept in Dhanyarashi (curing in Hordeum vulgare L.) for 7 days. During the procedure of Paka, temperature was maintained between 90°C and 100°C. On average, it took 8 h and 50 min to complete the process for ABR. A total of three batches of ABR were prepared; the average details of them are depicted in [Table 6] and [Figure 1].  | Figure 1: Graphic presentation of ABR preparation. ABR: Amrita Bhallataka Rasayana
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Preliminary analytical profile
The Ayurvedic Pharmacopoeia of India (API) standards were used for the physicochemical evaluation of raw and Shodhita Bhallataka. Raw and Shodhita Bhallataka, ABR was analyzed through relevant physicochemical parameters such as loss on drying (LOD),[11] ash value,[12] water-soluble extractive value,[11] alcohol‑soluble extractive value,[13] and pH value[14] in the modern pharmaceutical chemistry laboratory of the institute. Phytochemical qualitative analysis of the finished product was also carried out at the Institute. Nutritional evaluation and advanced high-performance thin-layer chromatography (HPTLC) of ABR were carried out.
| Results and Discussion | | |
Classics of Ayurveda have always emphasized the use of good quality drugs in therapeutics. They have provided specific instructions on how to determine whether a medicine is of good quality. They have suggested Bhallataka fruits that sunk in the water should be used in pharmaceutics and therapeutics. Most likely reason for this method may be the oil content in the Bhallataka fruits. Those Bhallataka fruits, which settle down in water, may have a higher percentage of oil content. The Bhallataka nut shell liquid present in the pericarp of the fruit contains tarry oil mainly comprising 90% of anacardic acid and 10% of cardol.[15] Pharmacognostical evaluation of Bhallataka showed the presence of palisade cells, parenchyma cells with fixed oil, and stone cell. Due to the presence of evacuated air-filled cells, Bhallataka fruits that float on the water surface may have less oil. During this examination, 78% of the yield was obtained when 2 kg of Bhallataka fruits was poured into the water. The color of the water turned to a light brown after dipping Bhallataka fruits.
Ayurvedic classics have clearly mentioned that drugs such as Bhallataka should be used after Shodhana (processing).[16] The role of Shodhana in poisonous plant materials such as Bhallataka has been well reported before their use for internal administration, and ancient Ayurvedic scholars are also well versed about this fact. In the current formulation, classic has advised Shodhana of Bhallataka with the use of brick powder. Brick powder has adsorbent property, by which it absorbs irritant oil in the fruit during Shodhana procedure.[17] It was reported that the oil content of Bhallataka fruits was decreased after 7 days of Shodhana in Ishtika Churna.[15] Tarry oil present in the pericarp of the fruit causes blisters on contact with other symptoms included itching, redness, burning sensation, swelling, papules, vesicles, blisters, and streaking.[18] It is the main reason behind Shodhana procedure of Bhallataka fruits.
Raw and Shodhita Bhallataka were subjected to pharmaceutical analysis using relevant physicochemical parameters as per API. This analysis was carried out to get genuinity and qualitative [Table 7] and quantitative composition of both samples. The results of physicochemical parameters of both raw and Shodhita Bhallataka are depicted in [Table 8]. Bhallataka fruits were hard and grayish-black in color before Shodhana and become soft, light in weight, and black after Shodhana. The color of brick powder was also changed to brownish-black color after the procedure. The results from after Shodhana of Ashuddha Bhallataka are summarized in [Table 9].  | Table 8: Average physicochemical analysis of raw and Shodhita Bhallataka
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In Bhallataka Kwatha preparation, raw fruits were crushed and broken down in coarse form as per the recommendation of classic.[19] It will facilitate the further process of extraction. The oil content was floated on the surface of the liquid media and the consistency of Kwatha became thick and sticky with dark brown color at the end of the procedure. The results of Kwatha preparation are summarized in [Table 10] and [Table 11]. | Table 10: Parameters and results obtained during the preparation of Bhallataka Kwatha
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Continuous stirring is necessary for proper extraction and to lessen the chances of degradation of some active constituents, which may be decomposed due to hydrolysis.[20] It also facilitates natural circulation evaporation.[21] Constant observation and continuous stirring are essential in obtaining a good quality of Kwatha and Avaleha particularly, during the initial stages of the procedure; otherwise, sugar in the central part will get caramelized.[22]
In ABR preparation, the exact quantity of sugar is not mentioned in the classics. To fix the ratio of sugar, four pilot batches of ABR were prepared with different proportions of sugar [Table 4]. The proper consistency of the formulation was achieved with the addition of equal amount of sugar and Kwatha material. In the final batch preparations, Godugdha, Goghrita, and powdered Sharkara were added to the Kwatha in the appropriate ratios with specific time intervals and were heated. Milk was added to the Kwatha after Cooling down at room temperature to avoid curdling as Bhallataka has Ushna-Tikshna properties. Goghrita was not mixed with Kwatha and milk mixture. It remains floating on the surface of liquid media. Milk products are sensitive to heat treatment encountered under conventional process because of an abundance of reactive functional groups. The most important heat-induced changes in dairy products that involve lactose are the changes associated with browning. The reaction of lactose with the caseins and whey proteins of milk systems may be via the Maillard or nonenzymatic browning reaction. It is an extremely complex process and is the reaction between reducing sugars and proteins by the impact of heat.[23]
During the preparation of ABR, after dissolving sugar in Kwatha, the color of the solution became darker, and a typical smell of sugar was observed during Paka. Heating was done for 15 min on 1st day, after the addition of a specified ratio of sugar to Kwatha mixture. The remaining process of Paka was done on the next day. The temperature variation with their chief desired characteristics at various time intervals during the preparation of ABR is noted [Figure 2]. After observing the desired characters such as Darvi Pralepa, Apsumajjanam, Patitastu Na Sheeryate, and Rasa Gandha Varnotpatti, the container was removed from the heat source and allowed to become cool. Thread-like consistency (Tantumatvama) was not seen in ABR preparation, which is common for most of all Avaleha formulations preparation, because it was directly converted into Khova (dried evaporated milk solids)-like consistency. Throughout the ABR preparation process, the temperature was maintained between 95°C and 110°C. [Table 12][Table 13][Table 14] described observations regarding timing and temperature during preparation and chief desired characteristics of Amrita Bhallataka Rasayana. The application of heat increases the solubility of sugar in the aqueous media. The solubility of sucrose is 75%–80% at the temperature of 90°C–100°C.[24] Color changes were observed throughout the process of ABR preparation [Figure 3]. The final product of ABR is of coffee brown color. Products of Maillard browning reaction include desirable and undesirable colors and aromas. The Maillard reaction causes products to brown by producing certain chemicals that are responsible for the browned product’s aroma and flavor. | Figure 2: Temperature and chief desired characteristics graph with time interval during the ABR preparation. ABR: Amrita Bhallataka Rasayana
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 | Table 12: Observations with timing and temperature during the preparation of Amrita Bhallataka Rasayana
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 | Table 13: Observations on the temperature at different stages of Amrita Bhallataka Rasayana preparation
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 | Figure 3: Variation of ABR color during various time intervals. ABR: Amrita Bhallataka Rasayana
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In the physicochemical analysis of ABR, LOD, ash value, water-soluble extractive value, alcohol‑soluble extractive value, and pH value were assessed. The average results of organoleptic and physicochemical analyses for three batches of ABR are summarized in [Table 15] and [Table 16]. pH of ABR showed that 10% w/v aqueous solution of the sample was slightly alkaline in nature ranging between 7 and 8. An alkaline pH may inhibit, or slow down, bacteria growth. The alkaline pH value of ABR may be observed due to protein-tannins complexes between milk and oil of Bhallataka. All three samples show very similar moisture content, whereas batch II has shown a bit lower moisture. However, a range between 28% and 31% w/w can be fixed as a desirable LOD. Total ash value helps to know about inorganic material in the test drug. A total of 0.56% of the ash value of ABR is found as it is a polyherbal formulation containing no minerals and metals. This indicates that ABR is purely organic in nature. The formulation is more bio-assessable to human biological systems due to the organic nature of the components. The total inorganic chemicals present in a product determine the ash value; this metric is important in drug quality control and standardization. The ash value of a drug will be lower if it contains more organic components. The average water-soluble extractive of ABR was 54.14%, whereas alcohol-soluble extractive was 19.93%. The extractive values of ABR were found more may be due to more quantity of herbal drugs present in it. There has been a changed leaching behavior of samples with water and methanol, which is probably due to matrix effect. As decoction of Bhallataka, Ghrita, sugar, and milk are ingredients of ABR, there is significant water-soluble extractive of the formulation. More alcohol-soluble extractive of ABR may be due to the fat content (ghee, milk, and Bhallataka Taila) of Avaleha.
Here, total reducing and nonreducing sugars were estimated. For the estimation of total sugar, the phenol-sulphuric acid method was followed, and for reducing and nonreducing sugar, Nelson’s Somagy’s method was followed. The total sugar values were obtained at 22.1% w/w in ABR. It was also observed that above 4.6% was reducing sugar and 17.5% was nonreducing sugar. In this formulation, total sugar was suggesting more than one-fourth part of the total contents and helps to improve the palatability of the formulation [Table 17].
In phytochemical qualitative analysis, the presence of saponins, flavonoids, and tannins was assessed using appropriate qualitative testing methods [Table 18]. The presence of functional groups and essential active ingredients of raw drugs in the finished product was detected using qualitative testing, indicating that these compounds were extracted in the formulation.
HPTLC was done to develop a chromatographic pattern of three samples of ABR by following the standard procedure at 254 and 366 nm [Table 19]. In a sample of ABR1, five spots were found at Rf values of 9.3, 11.4, 13.2, 7.6, and 15.6 in short ultraviolet (UV) at 254 nm [Figure 4], whereas two spots were found at Rf values of 5.7 and 22.3 at 366 nm [Figure 5]. In ABR 2 also, five spots were found at Rf values of 6.6, 12.6, 8.4, 12.9, and 10.2 in short UV at 254 nm [Figure 6], whereas three spots were found at Rf values of 0.09, 0.24, 0.29 at 366 nm [Figure 7]. ABR 3 sample also showed five spots at Rf values of 10.8, 2.3, 8.3, 1.8, and 10.3 in short UV at 254 nm [Figure 8], whereas two spots were found at Rf values of 12.4 and 25.3 at 366 nm [Figure 9]. There was a total of 0 similar peak found in all three ABR samples at 254 and 366 nm. Although in ABR 2 and ABR 3, nearby two similar peaks (8.4 and 8.3, and 10.2 and 10.3) were found at 254 nm. In short UV radiation, all three samples of ABR contain higher herbal material sensitivity [Table 20]. But in long UV, all three samples of ABR show lower sensitivity. It showed that the active contents in the formulation are more sensitive to short UV radiation (254 nm) compared with long UV radiation (366 nm).  | Figure 4: Graphs of HPTLC ABR I at 254 nm. ABR: Amrita Bhallataka Rasayana, HPTLC: high-performance thin-layer chromatography
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 | Figure 5: Graphs of HPTLC ABR I at 366 nm. ABR: Amrita Bhallataka Rasayana, HPTLC: high-performance thin-layer chromatography
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 | Figure 6: Graphs of HPTLC ABR II at 254 nm. ABR: Amrita Bhallataka Rasayana, HPTLC: high-performance thin-layer chromatography
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 | Figure 7: Graphs of HPTLC ABR II at 366 nm. ABR: Amrita Bhallataka Rasayana, HPTLC: high-performance thin-layer chromatography
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 | Figure 8: Graphs of HPTLC ABR III at 254 nm. ABR: Amrita Bhallataka Rasayana, HPTLC: high-performance thin-layer chromatography
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 | Figure 9: Graphs of HPTLC ABR III at 366 nm. ABR: Amrita Bhallataka Rasayana, HPTLC: high-performance thin-layer chromatography
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The test drug was screened for qualitative and quantitative analyses of some nutritional value parameters such as total proteins, nitrogen, total fat, total crude fibers, and total carbohydrates. The results of the nutritional analysis are presented in [Table 21]. The presence of total protein (4.42%) is due to milk. Total fat (33.61%) is present because of milk and ghee. The presence of total carbohydrates (36.06%) is more because of sugar and milk.
| Conclusion | | |
No significant changes were found in physicochemical aspects of raw and Shodhita Bhallataka. The method of preparation given in the current study for ABR may be refered for standardization purpose. Parameters of ABR are not mentioned in the Ayurvedic Pharmacopeia of India. Hence, these preliminary parameters may be helpful as standards for further studies. This study may provide certain leads toward using different proportions of sugar in the preparation of ABR. However, extensive studies focusing on the exact mechanism pharmacodynamically and pharmacokinetically be carried out, and changes that take place with the change in sugar proportion to emphasize its rationale and impact are needed.
Financial support and sponsorship
Nil.
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]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7], [Table 8], [Table 9], [Table 10], [Table 11], [Table 12], [Table 13], [Table 14], [Table 15], [Table 16], [Table 17], [Table 18], [Table 19], [Table 20], [Table 21]
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