|Year : 2022 | Volume
| Issue : 2 | Page : 140-146
Insight into the Ayurveda theory of “Dravyasamgrahakala” on Shatavari (Asparagus racemosus Willd) root with reference to quality standard parameters
Kinjal Parsotam Unadkat1, Dilip Jani1, Hari Om Gupta2, Neha Parmar3
1 Department of Dravyaguna, Government Ayurveda College, Vadodara, Gujarat, India
2 Department of Dravyaguna, Government Ayurveda College, Junagadh, Gujarat, India
3 State Model Institute of Ayurveda Sciences, Gandhinagar, Gujarat, India
|Date of Submission||03-Apr-2021|
|Date of Decision||20-Apr-2021|
|Date of Acceptance||26-Dec-2021|
|Date of Web Publication||4-Jul-2022|
Kinjal Parsotam Unadkat
Assistant Professor, Department of Dravyaguna, Noble Institute of Ayurveda & Research, Bamangam, Junagadh, Gujarat
Source of Support: None, Conflict of Interest: None
Introduction: India has been known to be the richest heritage of medicinal plants. Globally, there is a universal rise in the use of Ayurveda system of medicine. According to the WHO, it has been evaluated that 80% of world's population relies on traditional medicines. There are many factors such as geographical variations, place, habitat, season, and temperature can affect the quality of medicinal plants. The study is an endeavor to compare the effect of seasonal variations on plants on the basis of quality standard parameters to validate the basic principle of Ayurveda “Dravyasamgraha kala.” This study aims to evaluate analytical changes and compare the data of different samples of Asparagus racemosus Willd. collected in different seasons according to Ayurveda classics. Methods: In Ayurveda texts, there is also a description about parts of the plants that should be collected in specific seasons. For the purpose, Shatavari (A. racemosus Willd.) was selected for the study. In the current research, four samples of roots of A. racemosus were collected in four different seasons-Grishma, Pravrut, Sharad, and Shishiraritu (May, June, October, and January) to compare the data and find out whether there is any similarity or difference in analytical profile. Results and Conclusion: The quality difference including physicochemical parameters, phytochemical identification, and high-performance thin-layer chromatography was remarked in the samples. It can be concluded that chemical constituents make a unique revelation from different seasons. Proper time period of the collection of plant is very important to obtain good quality drug. The season has a significant role to validate the factors that influence the potency of the drug.
Keywords: Analytical study, Asparagus racemosus, Ayurved, Dravyasamgraha kala, Shatavari
|How to cite this article:|
Unadkat KP, Jani D, Gupta HO, Parmar N. Insight into the Ayurveda theory of “Dravyasamgrahakala” on Shatavari (Asparagus racemosus Willd) root with reference to quality standard parameters. J Ayurveda 2022;16:140-6
|How to cite this URL:|
Unadkat KP, Jani D, Gupta HO, Parmar N. Insight into the Ayurveda theory of “Dravyasamgrahakala” on Shatavari (Asparagus racemosus Willd) root with reference to quality standard parameters. J Ayurveda [serial online] 2022 [cited 2022 Aug 10];16:140-6. Available from: http://www.journayu.in/text.asp?2022/16/2/140/349773
| Introduction|| |
India has a very rich plant biodiversity worldwide. The medicinal plants are known to be the main source of traditional medicines, phytopharmaceuticals, nutraceuticals, many of modern allopathic drugs, household remedies, and cosmetics with a very high degree of societal acceptance. In the current scenario, people are getting more attracted toward Ayurveda therapies which are mainly plant based. Demands of medicinal plants are increasing day by day. Therefore, it is necessary to create a need for conserving medicinal plants. Overexploitation of the medicinal plants is leading to unsustainable collections of the plants from the forests which are the main source of raw material used in Ayurveda medicines resulting in unavailability and disappearing of the medicinal plants at a large rate. To prevent deterioration of ecosystem through ruthless exploitation of medicinal plants, one should plan to collect the drug with the highest pharmacotherapeutic activity. Thus, the efficacy of the drug will be increased and acquired prompt result will be appointed. Ayurvedic classics such as Charaka samhita, Sushruta samhita have documented information regarding collection of parts of the plants according to season [Table 1] and [Table 2]. The herbal drugs are in sync with nature which is the biggest advantage. The season is one of the factors can affect the potency of the drug. Thus, medicinal plants should be collected in a specific season. For the purpose of evaluation, the drug Shatavari (Asparagus racemosus Willd.) was selected in this study, and the samples were collected in different seasons as per the references of Ayurveda texts. The Ayurvedic Pharmacopoeia of India was introduced to establish and ensure certain minimum quality aspects of medicinal plants. Such studies can help to explore the reasoning and concept behind the classical principles related to drug collection. It is possible only after the observation and systemic study. In the current research, it was demonstrated that when the drug was collected in specific season, as per mentioned in classics, the changes in quality standard parameters such as physicochemical analysis, phytochemical analysis, and qualitative analysis were observed according to season. So here, an attempt has been made to establish and prove the fundamental concept of Ayurveda “Dravyasamgraha kala.”
|Table 1: The description of collection of parts of the plants in different seasons according to classics|
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|Table 2: Description of season with their corresponding Hindu Lunar month, Gregorian month and origin of Rasa|
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Aims and objectives
The study of the samples of roots of A. racemosus was undertaken with the following aims and objectives:
- To evaluate the samples of Shatavari roots collected in different seasons using different physicochemical parameters and qualitative methods
- To compare the data and to find out whether there is any similarity or difference in analytical profile of the samples collected in different seasons.
| Materials and Methods|| |
The sample drug plant (A. racemosus) was propagated in 10 clay pots at Dhanvantari garden of State Ayurved Faculty, Vadodara. In Sushruta samhita, it is mentioned that the drug ought to be used when its aged 1 year old. Hence, the first sample was collected for the study after 1 year. The sample drug plant (A. racemosus) was propagated in 10 clay pots within May 2018. Then, when 1 year (May 2019), the primary sample was collected and documented by evaluators [Figure 1]. In the present study, the samples of roots of A. racemosus were collected in four different seasons-Grishma, Pravrut, Sharada, and Shishira ritu on the basis of Ayurveda documented literature [Table 1].
|Figure 1: Microscopical characters of Asparagus racemosus. (a) T.S. of the root. (b) Cork cells. (c) Piliferous layer. (d) Cortex. (e) Raphides of calcium oxalate. (f) Xylem and phloem|
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Then, the samples were washed and dried in shade for 40–45 days in four different seasons. The dried pieces of Shatavari roots were converted into a coarse powder with mixture grinder and sieved through 80 size mesh. The samples of the roots were preserved in air-tight containers for analytical study. All analytical tests were done at Vasu Research Centre, Makarapura, Vadodara.
The analytical work was administered in the following parts:
- Physicochemical analysis
- Preliminary phytochemical investigation
- High-performance thin-layer chromatography (HPTLC) profile.
- Loss on drying (%w/w)
- Total ash (%w/w)
- Acid insoluble ash (%w/w)
- Water soluble extractive (%w/w)
- Alcohol soluble extractive (%w/w)
- pH (1%).
Determination of loss on drying
- A shallow glass stopper weighing bottle was taken that had been dried for 30 min below a similar condition to be used within the determination and cooled in a desiccator. Then, accurately weigh 1 g (note the exact weight) of the sample was uniformly mixed and transferred it to the bottle, and immediately, the lid was closed
- Then the stopper bottle was weighed with the sample in it, and the exact weight (A) was noted. Then, the loaded bottle was arranged in the drying chamber. The stopper was removed and left it also in the chamber. The sample was dried at 105°C ± 2°C for about 1.0 h. At the end of 1 h, the drying chamber (Hot air oven) was opened and immediately the lid was placed back on the bottle
- The bottle was transferred back into the desiccator and allowed it to cool. Once the bottle was cooled then weighed the bottle and the exact weight (B) were noted. The percentage of value of loss on drying was calculated using the formula:
Determination of total ash
- The air-dried sample of test drug was accurately weighed 1.0 g (note the weight) and transferred it to a pre-weighted crucible. Then, the crucible with the sample was weighed, and the precise weight was noted
- The sample was incinerated gently at 450°C till free from carbon. If a carbon-free ash cannot be obtained in this way, cool the crucible, exhaust the burn mass with hot water, collect the residue on an ash-less filter paper, burn the remnant and filter paper, include the filtrate, evaporate to dry, and burn at a temperature not exceeding 650°C
- The crucible was removed from the furnace after that, it was cooled in desiccators. Then, the crucible was weighed. The difference in the weight is the weight of the ash. The total ash content in the sample was calculated using the formula:
Determination of acid insoluble ash
- The ash obtained from total ash was taken to determine the acid insoluble ash content. Then, 25 mL of 10% dilute HCl was added and boiled on water bath for 10 min. It was filtered through ash-less filter paper. The insoluble matter was washed with hot water. The insoluble matter of ash was transferred with filter paper in the earlier taken crucible and dried it on hot plate
- It was ignited for 1 h at 800°C in furnace. The crucible was removed from the furnace. It was cooled in desiccators. Then, the weight of crucible was considered.
- The acid-insoluble ash content was calculated using the formula:
Determination of water soluble extractive
- The same method was followed as above described to determine acid insoluble ash
- The insoluble matter of ash was ignited for 20 min at 650°C in furnace. The crucible was removed from the furnace and cooled in desiccators. Then, the weight of crucible was noted
- The water-soluble ash content was calculated in the sample using the formula:
Determination of alcohol soluble extractive
- The sample was accurately weighed 5.0 g (note the exact weight) of and transferred it to a 250 mL conical flask. Then, 100 mL of alcohol was added to this. The flask was kept on the shaker for 6 h and then allowed to stand for 18 h. It was filtered
- Then, 25 mL of the filtrate was evaporated to dryness in weighed evaporating dish. Further, it was dried at 105°C to a constant weight and weigh
- The percentage of alcohol-soluble extractive was calculated using the formula:
Determination of pH
- Accurately weighed 1 g of sample was taken and transferred it to a 100 ml of capacity beaker. Then, 100 ml of water was added to this and shaken well for about 1.0 min. The pH meter electrode was put in the beaker. The reading was noted and then washed the rod 2–3 times with distilled water.
The preliminary analysis of phytochemical composition is one in all the most effective strategies to analyze quality of herbal drugs. The phytochemical analysis of the samples such as alkaloids, flavonoids, tannins, and saponins was remarked in four different seasons.
Preparation of plant extracts
Five grams of each samples of Shatavari root collected in four different seasons, i.e., Grishma (May 2019), Pravrut (June 2019), Sharad (October 2019) and Shishira (January 2020) were extracted with methanol (100 ml), keeping it for nightlong with initial occasional shaking up to 6 h and then set aside. It had been filtered and alcoholic extract was collected once 24 h. Similarly, water extracts of both samples were prepared and collected. The following qualitative tests were done by using appropriate extracts.
The extract was dissolved in alcohol. Then, a couple of drops of acetic acid and Dragendroff's chemical reagent were added and shaken well. If there is an orange–red precipitate found, it suggests that there is the presence of alkaloids.
The sample was dissolved in 10%HCL and added Zinc (dust). It had been determined the effervescence with pink color indication of flavanones, flavanol, and also the glycosides. When it was treated with neutral lead acetate gives yellow, orange, red, or brick color precipitation. Flavonoids were dissolved in alkali with the formation of yellow solution which undergoes rapid color changes on standing for some time on the addition of few drops of HCl the color was disappeared. Flavonoids were dissolved in H2SO4 to give intensely yellow solution.
Tannins are soluble in water, acetone, and alcohols, glycerol, and they may be precipitated with ferrous salts. When aqueous extract of the sample was treated with very dilute solution of ferric chloride, there will be ever-changing of blue color to olive green.
In alcohol extract, two drops of alcoholic ferric chloride were added. If there is blue to blue–black formation found, it exhibits the presence of phenol.
The extract was dissolved in chloroform. Then, 1 ml of acetic acid and 1 ml of acetic anhydride were added. After that, it had been heated on a water bath and cooled. Few drops of concentration of sulfuric acid were added along the sides of the test tube. If bluish-green color appearance found, it shows the presence of steroids.
To a few mg of extract was taken. In this, distilled water was added and shaken. If there is stable froth formation found, it indicates the existence of saponins.
The results are depicted in tabular manner() for detail comparative analysis of the samples.
High-performance thin-layer chromatography profile
HPTLC is a very popular and powerful method for analysis. It is an automated, advanced, ultra-modern, and upgrade methodology of TLC. With using the method, a similar analysis may be viewed using different wavelengths of light. Therefore, it offers broad primarily based profile of the plant and additional specific kinds of analysis. The method is equally appropriate for each qualitative also as quantitative analysis of the drugs.
HPTLC is that the separation technique of the elements of a mixture and is due to
- Their totally different affinities for a stationary phase like solid or liquid and
- Their differential solubility in a very moving phase like a liquid or gas.
Steps involved in high-performance thin-layer chromatography
- Choice of chromatographic layer
- Sample and standard preparation
- Layer prewashing
- Layer preconditioning experimental study
- Application of sample and standard
- Chromatographic development
- Detection of spots
- Scanning and documentation.
Samples for high-performance thin-layer chromatography
The samples of Shatavari roots were collected in four totally different seasons, i.e., Grishmarutu (May 2019), Pravrut rutu (June 2019), Sharad rutu (October, 2019), and Shishira rutu (January 2020).
Preparation of test solutions
Accurately weighed 2 g sample was taken in Iodine flask and 20 ml methanol was added to it. It had been then refluxed on water bath for 30 min and allowed to cool. Concentrate up to 1 mL and then make up the volume up to 10 mL.
Preparation of standard solutions(s)
Accurately 1 mg of standard Shatavarin-IV into 1 mL volumetric flask was weighed and dissolved in methanol (1 mL).
- Applicable Mode: CAMAG Linomat five – Applicator
- Filtering System: Whatman paper No. 1
- Stationary Phase: MERCK– TLC/HPTLC Silica gel 60254 on Aluminum sheets
- Application (Y axis) Start Position: 10 mm
- Development (Y axis) End Position: 80 mm from plate base
- Space Between Band: 10 mm
- Sample Application Volume: 5 μL
- Standard Application volume: 10 μL
- Development Mode: CAMAG TLC Twin Trough Chamber
- Chamber saturation time: 35 min
- Mobile Phase (MP): Toluene: Ethyl Acetate: Methanol: Water (7.5:1.5:1)
- Visualization: @ 429 nm
- Spray reagent: Anisaldehyde Sulfuric acid reagent
- Derivatization mode: CAMAG– Dip tank for regarding 1 min
- Drying Mode, Temp. and Time: TLC Plate Heater Preheated at 100°C ± 5°C for 3 min
- Observation: Plate was examined at 429 nm under ultraviolet (UV) cabinet for the appearance of different bands at different Rf.
| Results and Discussion|| |
The roots of Asparagus raemosus were collected in four different seasons quoted by varied Acharyas to discover which season is the best for collection [Table 1]. The study was carried out in the phases such as physicochemical analysis, phytochemical identification, and HPTLC profile. The result of the study with discussion is as follows [Table 3], [Table 4], [Table 5], [Table 6], [Table 7]:
|Table 6: High performance thin layer chromatography profile of the samples of Shatavari root|
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|Table 7: High performance thin layer chromatography Assay of the samples of Shatavari root|
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The value of loss on drying was found lowest (5.33% w/w) in first sample collected in Grishma ritu as mentioned by AcharyaCharaka. Thus, it may be said that first sample may be considered as good enough in shelf life and quality material. The Grishmaritu may also have the low moisture naturally.
Ash value influences the quality and purity of the plants. Ash value is also found higher in first sample (5.90% w/w). According to higher ash value, the preferable collection time period of the drug is Grishmaritu which is related with the theory of Charaka.
The values of water-soluble extractive (58.66% w/w) and alcohol soluble extractive (67.02% w/w) were found higher in third sample collected in Sharad ritu with the references of Sharangadhara samhita, Bhavaprakasha samhita, and Nighantu ratnakara. It can be justified that collected samples in Sharada ritu have more water solubility as compared to others. If we need hydroalcoholic extract, the best season is Sharada ritu.
The higher quantity of saponin was also observed in third sample which was collected in Sharada ritu. It shows that the quantity of saponin may be influenced by the seasons. It might be that Sharada ritu is a suitable season for more production of saponin in Shatavari. Thus production of T cells may be highly stimulated by saponin in Sharada ritu. It can be said that one can get more advanced immunomodulatory and antioxidant response from the drug containg saponin in Sharada ritu. Hence, the drug should be collected in Sharada ritu with special reference to saponins [Figure 2].
|Figure 2: Peak display of densitogram at 429 nm. (a) 1st sample: Grishma ritu. (b) 2nd sample: Pravrit ritu. (c) 3rd sample: Sharad ritu. (d) 4th sample: Shishira ritu|
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HPTLC was observed under UV cabinet 429 nm, the maximum area under curve was reported in third sample collected in Sharad ritu as per the references of Sharangadharasamhita, Bhavaprakasha samhita, and Nighantu ratnakara compared to the other samples. It is inferred that the Sharada ritu is the best season to collect root with special reference to Shatavarin IV.
Overall, it is concluded that water-soluble extractive, alcohol soluble extractive, HPTLC assay, and saponin content are observed highest in third sample (Sharada ritu). It shows that quantity of saponins, HPTLC assay, and extractive values may be influenced by the seasons. Hence, technically and logically, the third sample is considered to be the best with special reference to chemical compositions and study available till date based on this analysis. The theory put on by Sharangadhara samhita, Bhavaprakasha samhita, and Nighantu ratnakara seems to be justified and correct based on the current study.
The authors are thankful to the AYUSH, Gujarat state, and all the faculties who have directly or indirectly helped to make this literature.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Sharangadhara. Sharangadhara Samhita, Purvakhanda, Prathamodhyaya, Reprint. Ch. 1., Ver. 67. Varanasi: Chaukhambha Orientalia; 2015. p. 15.
Bhavamishra. Bhavaprakasha Samhita, Purvakhanda, Mishraprakarana, Reprint 12th
Edition. Ch. 6., Ver. 91-97.Varanasi: Chaukhambha Sanskrit Bhavana; 2015. p. 181-2.
Navare K. Nighantu Ratnakara. 1st
ed. Delhi: Chaukhambha Sanskrit Pratishthan; 2011. p. 293.
Anonymous. The Ayurvedic Pharmacopoeia of India. Part 1.Vol. IV. 1st Edition. Government of India, New Delhi: Ministry of Health and Family Welfare, Department of Ayurveda, Yoga- Naturopathy, Unani, Siddha and Homeopathy (AYUSH); 1999. p.109.
Maes M, Stevens W, Scharpé S, Bosmans E, De Meyer F, D'Hondt P, et al.
Seasonal variation in peripheral blood leukocyte subsets and in serum interleukin-6, and soluble interleukin-2 and -6 receptor concentrations in normal volunteers. Experientia 1994;50:821-9.
Cao Y, Fang S, Fu X, Shang X, Yang W. Seasonal variation in phenolic compounds and antioxidant activity in leaves of Cyclocarya paliurus
(Batal.) Iljinskaja. Forests 2019;10:1-17.
[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7]