HPTLC Standardization of an Ayurvedic Polyherbal Tablet for Covid-19 Management

 

Rajasekaran A*, Mohanapriya B, Arivukkarasu R

KMCH College of Pharmacy, Department of Pharmaceutical Analysis, Coimbatore, Tamil Nadu, India.

*Corresponding Author E-mail: arsekaran@gmail.com

 

ABSTRACT:

The present study was aimed to scientifically validate the ayurvedic polyherbal tablet formulation used for the treatment of COVID-19. Organoleptic evaluation, physiochemical evaluation such as water-soluble extractive value, alcohol soluble extractive value, pH determination, weight variation, disintegration test, phytochemical investigation and HPTLC analysis, determination of microbial contamination and heavy metal analysis of the Ayurvedic Polyherbal Tablet Formulation were studied. The HPTLC analysis of Ayurvedic Polyherbal Tablet Formulation confirmed 0.04% of rutin in each methanol and aqueous extracts, 0.46% and 0.14% of quercetin, 0.04% and 0.17% of gallic acid, 0.19% of piperine in each methanol and aqueous extracts, 0.11 and 0.32% of andrographolide in methanol and aqueous extracts respectively. Except weight variation, remaining evaluation tests such as extractive value, disintegration time, pH and the phytochemical investigation of Ayurvedic Polyherbal Tablet Formulation were found to be within the pharmacopeial limits. HPTLC investigations of methanol and aqueous extracts of the Ayurvedic Polyherbal Tablet Formulation confirmed the presence of all ingredients mentioned in the Label claim. Microbial tests and heavy metal analysis revealed that the formulation is safe as it does not contain any microorganisms or heavy metals.

 

KEYWORDS: HPTLC Analysis, Ayurvedic Polyherbal Formulation, COVID-19 Management, Phytochemical Evaluation, Standardization.

 

 

 

INTRODUCTION:

Ayurveda - the science of life, evolved as a comprehensive system of healthcare systematically through scientific experimentations of high order backed by sound and reproducible evidence base and stood the test of the time1. The Ayurvedic Polyherbal Tablet Formulation consists of ten ingredients which includes Andrographis paniculata, Carica papaya, Cyperus rotundus, Melia azedarach, Mollugo cerviana, Piper nigrum, Tinospora cordifolia, Tricosanthus dioica, Vetiveria zizaniodes and Zingiber officinale. The polyherbal formulation was primarily developed to treat dengue, has been repurposed as supportive treatment for mild to moderate COVID-19 cases. It is reported to be safe. This formulation is proven to be effective as an analgesic, antipyretic and reversal of thrombocytopenia. It is absolutely safe for people with liver or kidney ailments and can be safely co-administered with other drugs. It can also be used as a prophylactic treatment for primary contacts of COVID-19 positive patients and frontline healthcare workers. The efficacy profile of the formulation were studied in detail and substantiated with scientific evidences for the benefits of the medicinal plants. Uma Narayanamurthy evaluated the acute and repeated dose toxicity effects of this Polyherbal syrup formulation prepared from ten different herbs used in traditional medicine for the management of viral infections and other inflammatory disease conditions2. This present study on the HPTLC standardization of an Ayurvedic polyherbal tablet for COVID-19 management is a continuation of our earlier research3 on the Siddha formulation Kabasura Kudineer against SARS-CoV-2. Both formulations share a significant overlap in their composition, as they include traditional medicinal plants with well-documented antiviral and immunomodulatory properties. Notably, Andrographis paniculata has demonstrated potential in inhibiting viral replication and modulating immune responses, as highlighted in the earlier molecular docking study. The fruit peel of Carica papaya has been investigated and reported to possess significant antioxidant and antimicrobial properties, which may contribute to its observed antiviral activity, as oxidative stress modulation and microbial inhibition are often linked to enhanced antiviral defense mechanisms4. Similarly, Piper nigrum is renowned for its bioavailability-enhancing properties and antiviral activity. Zingiber officinale exhibits anti-inflammatory and antiviral effects, contributing to the alleviation of respiratory infection symptoms. The phenolic extract from the rhizome of Zingiber officinale (ginger) exhibits potent antioxidant activity, which may contribute to its antiviral potential by mitigating oxidative stress-induced cellular damage and enhancing host immune response - mechanisms that play a critical role in antiviral defense5. Cyperus rotundus is a traditional herb valued for its immune-boosting and anti-inflammatory properties. The high total phenol and tannin content observed in Cyperus rotundus may contribute to its medicinal significance, particularly its antiviral activity, as these polyphenolic compounds are known to inhibit viral replication, interfere with viral enzymes, and enhance host defense mechanisms through antioxidant and immunomodulatory actions6. The antioxidant as demonstrated in rat models, may contribute to its antiviral potential, by reducing oxidative stress and improving lipid metabolism, the extracts help maintain cellular integrity and immune balance - conditions that are favorable for inhibiting viral entry, replication, and associated inflammatory responses7. The novel therapeutic potential of Mollugo cerviana may extend to antiviral applications, as its bioactive constituents are reported to exhibit antioxidant, anti-inflammatory, and immunomodulatory properties - mechanisms that are critically involved in suppressing viral replication and enhancing host antiviral defense8. Piper nigrum exhibits antipyretic activity by inhibiting prostaglandin synthesis through COX enzyme modulation, reducing fever and inflammation. Its antiviral activity is linked to the inhibition of viral entry and replication by disrupting viral envelope proteins and enhancing host immune response. Both effects are attributed to active constituents like piperine, which modulate immune and inflammatory pathways9. Tinospora cordifolia is widely recognized for its immunomodulatory and adaptogenic properties, which play a pivotal role in combating infections10. Carica papaya exhibits antiviral, antifungal, and antimicrobial activities through phytochemicals like papain, flavonoids, and alkaloids present in its leaves, seeds, and latex. These compounds disrupt viral replication, inhibit fungal cell wall synthesis, and damage bacterial membranes via oxidative stress Each part acts through multi-targeted bioactive mechanisms, offering broad-spectrum defense11. Finally the aerial parts of Andrographis paniculata exhibit both antiviral and antibacterial activities due to the presence of andrographolide and related diterpenoids. These compounds inhibit viral protein synthesis and replication while also disrupting bacterial cell walls and interfering with DNA gyrase. Their dual mechanism enhances immune response and blocks pathogen growth effectively12. These attributes may also contribute to its antiviral potential by enhancing host immune responses, regulating cytokine production, and inhibiting viral replication through immune system modulation. Together, these shared constituents underscore the pharmacological potential of both formulations, bridging traditional medicinal practices with modern scientific validation for COVID-19 management.

MATERIALS AND METHODS:

Collection of herbal raw materials and tablet formulation:

The herbal raw materials such as Andrographis paniculata, Carica papaya, Cyperus rotundus, Melia azedarach, Mollugo cerviana, Piper nigrum, Tinospora cordifolia, Trichosanthes dioica, Vetiveria zizanioides, Zingiber officinale were procured from the local market of Madurai, India. The marketed Ayurvedic Polyherbal Tablet Formulation was procured from the Apollo Pharmacy, Coimbatore, India.

 

Chemicals and Instruments:

The standard markers used such as andrographolide, piperine, rutin, quercetin and gallic acid were purchased from Sigma Aldrich Chemical Private Limited, Mumbai. The solvents used are purchased from Qualigens Fine Chemicals Private Limited, Mumbai. A CAMAG HPTLC system comprising of a Linomat-5 applicator and CAMAG TLC Scanner-3 and single pan balance of Shimadzu model was used for the spotting.

 

Table 1 Physiochemical analysis of ayurvedic polyherbal tablet formulation

Parameters

Test sample 1

Test sample 2

Test sample 3

Average

Alcohol extractive value

4.4

5.2

5.8

5.1

Water extractive value

6.4

6.8

5.8

6.3

pH

8.15

8.20

8.26

8.20

Disintegration test

8 minutes 30 seconds

9 minutes 48 seconds

8 minutes 40 seconds

8 minutes 73 seconds

Weight variation

0.10 % to 1.73 %

 

Organoleptic Evaluation:

Color, odour and taste of tablet formulation was studied.

 

Physiochemical Investigation:

Determination of Alcohol Soluble Extractive Value:

Five g of air dried coarse powder was added to 100ml of alcohol in a closed flask for twenty-four hours and shaken frequently for six hours and allowed to stand for eighteen hours. Then filtered rapidly and 25ml of the filtrate was evaporated to dryness in a tared flat bottomed shallow dish, and dried at 105°C, to constant weight and weighed. Calculated the percentage of alcohol soluble extractive with reference to the air-dried drug.

 

Determination of Water Soluble Extractive Value:

Five g of air dried coarse powder was added to 100ml of distilled water in a closed flask for twenty-four hours and shaken frequently for six hours and allowed to stand for eighteen hours. Then filtered rapidly and 25ml of the filtrate was evaporated to dryness in a tared flat bottomed shallow dish, and dried at 105°C, to constant weight and weighed. Calculated the percentage of water soluble extractive with reference to the air-dried drug13.

 

Determination of pH:

The pH of the formulation was determined using ELICO pH meter with combined electrodes pH glass electrode and calomel reference electrode14.

 

Weight Variation:

Twenty tablets were individually accurately weighed. Each tablet weight was recorded. Results were reported as mean ± standard deviation in milligrams (mg) units. By randomly selecting and weighing 20 tablets, “the average weight was determined15.

 

Disintegration test:

Disintegration test was performed in a basket rack assembly. Distilled water was used as the medium and a disc was added to each tube. Operated the apparatus for 30 minutes. The limit is not more than 15 minutes for film coated tablets16 (Table 1).

 

Determination of Microorganisms:

The microorganisms such as bacteria and fungi were determined using spread plate technique. Petri dishes 9-10cm in diameter was used. Liquefied casein-soybean digest agar was added at a temperature not exceeding 45°C for bacteria. For fungi, sabouraud glucose agar was used17.

Determination of Heavy Metals:

Heavy metals present in Ayurvedic Polyherbal Tablet Formulation were detected using ICP-MS (Inductively Coupled Plasma Mass Spectrometer) after digestion with nitric acid18.

 

High Performance Thin Layer Chromatography Finger Print Profile:

Preparation of Samples Standards and Extracts from Raw Materials:

HPTLC study of methanol extract of the individual ingredients and methanol and aqueous extracts of the marketed formulation were carried out along with the different marker compounds corresponding to the active ingredients to ensure the presence of active ingredients in the marketed Ayurvedic Polyherbal Tablet Formulation. For HPTLC, 10g of each ingredients in 100ml of methanol was sonicated, filtered and then concentrated on boiling water bath for 20minutes. One g of tablet sample was extracted with 10ml distilled water. Likewise, 1g of tablet sample was extracted with 10ml methanol. The standard markers were prepared using 10 μg in 10ml methanol. The chromatogram was performed by spotting standards and extracted samples on pre coated silica gel aluminium plate 60GF254 using Camag Linomat V sample applicator. Plates were developed using suitable mobile phases. Subsequent to development, TLC plates were dried and densitometric scanning was performed on Camag TLC scanner in the absorbance mode at 245nm and 366nm.

 

RESULTS:

Weight variation of the tablets were within the range of 0.10 to 1.73%. The disintegration time of the film-coated ayurvedic tablet was found to be 8 minutes and 73 seconds, which is within the Indian Pharmacopoeia limit of not more than 15 minutes.

 

The aqueous and alcohol extractive values of the ayurvedic polyherbal tablet were determined to be 6.3% and 5.1%, respectively, indicating a higher content of water-soluble constituents. The pH of the tablet formulation was 8.20, reflecting a slightly basic nature.

 

Since, ayurvedic polyherbal tablet formulation composed of flavonoids, alkaloids, saponins, tannins, amino acids etc., it was not possible to separate the compounds using single mobile phase technique. The following different mobile phase composition were attempted to elute the components using HPTLC for the Ayurvedic Polyherbal Tablet Formulation. The mobile phase was optimized for procured raw materials of the ayurvedic polyherbal tablet formulation viz. Piper nigrum, Andrographis paniculata and a well resolved peaks were observed for the major constituents. The optimized mobile phase system comprising of Ethyl acetate: n-hexane in the ratio (8.5:1.5) well resolved the possible marker andrographolide in methanol and aqueous extract of ayurvedic polyherbal tablet and methanol extract for Andrographis paniculata. The overlay spectrum of the standard Andrographolide with methanol and aqueous extract of ayurvedic polyherbal tablet and Andrographis paniculata methanol extract corroborated the same (Fig. 1-2).

 

 

Fig.1 - Correlation of Rf value and λ max for methanol extract of Andrographis paniculata, methanol and aqueous extract of sample and standard and rographolide at 254 nm

T1-Methanol extract of sample, T2-Aqueous extract of sample, T3-Standard Andrographolide, T4-Methanol extract of Andrographis paniculata

 

Fig.2 - Densitogram of a) methanol extract of sample b) aqueous extract of sample c) standard andrographolide d) methanol extract of Andrographis paniculata at 254 nm

 

HPTLC analysis confirmed the presence of andrographolide (0.11% in methanol extract and 0.32% in aqueous extract, consistent with our earlier findings19  and piperine (0.19%). Flavonoids such as gallic acid, rutin, and quercetin were also identified, with their respective concentrations being 0.04%, 0.04%, and 0.46% in methanol extracts and 0.17%, 0.14%, and 0.32% in aqueous extracts20.The mobile phase optimized for Piper nigrum using Toluene: Ethyl acetate (7:3) resolved the components present in methanol and aqueous extract of ayurvedic polyherbal tablet and Piper nigrum methanol extract. The overlay of standard Piperine with methanol and aqueous extract of ayurvedic polyherbal tablet and Piper nigrum methanol extract confirmed the presence of piperine in the herbal formulation (Figure 3 and 4).

 

T1-Methanol extract of sample, T2-Aqueous extract of sample, T-3-Piperine standard, T4-Methanol extract of Piper nigrum

 

Fig. 3 - Correlation of Rf value and λ max for methanol extract of Piper nigrum, methanol and aqueous extract of marketed tablet sample and standard piperine at 254 nm

 

Fig.4 - Densitogram of a) methanol extract of sample b) aqueous extract of sample c) standard piperine   d) methanol extract of Piper nigrum at 254 nm

 

 

The HPTLC analysis of methanol extract and aqueous extract of Ayurvedic Polyherbal Tablet Formulation confirmed 0.19% of piperine respectively. Piperine has high binding affinity towards the RNA binding pocket of the nucleocapsid, thereby inhibiting viral proliferation. Hence, it is suggested that consumption of this piperine helps to combat COVID-19 directly through possible antiviral effects21. The mobile phase which comprising of Toluene: Ethyl acetate: Formic acid: Methanol in the ratio of 3:6:1.6:0.4, efficiently resolved the flavonoid components present in the tablet formulation22. The optimized chamber saturation time for mobile phase was 3 mins at room temperature (25 ± 1°C). The densitometric analysis was performed at 254 nm in reflectance mode. The presence of flavonoids such as gallic acid, rutin and quercetin were identified in methanol and aqueous extracts of ayurvedic polyherbal tablet. The HPTLC analysis of methanol extract and aqueous extract of Ayurvedic Polyherbal Tablet Formulation confirmed 0.04% and 17% of gallic acid respectively (Figure 5).

 

T1- methanol extract of sample, T2-aqueous extract of sample T3-Standard Rutin, Gallic acid, Quercetin, T4-Cyperus rotundus methanol extract, T5-Mollugo cerviana methanol extract, T6-Zingiber officinalis methanol extract, T7-Andrographis paniculata methanol extract, T8-Piper nigrum methanol extract, T9-Carica papaya methanol extract, T10-Vettivera zizannoides methanol extract, T11-Tinospora cordifolia methanol extract

 

Gallic acid is a COVID-19 RNA dependent RNA polymerase inhibitor23. Anti-oxidative, anti-inflammatory, anti-mutagenic antimicrobial, free radical scavenging abilities and other medicinal properties coupled with their capacity to modulate essential cellular enzyme functions are mainly due to the presence of flavonoids. The HPTLC analysis of methanol extract and aqueous extract of Ayurvedic Polyherbal Tablet Formulation confirmed 0.04% of rutin. Rutin has been identified as a potential hit, having prominent binding affinity to the virus. Rutin is an effective inhibitor of main protease enzyme which is present in COVID-19, thus preventing the replication of the virus. Its presence in traditional medicines prescribed to infected patients with mild to moderate symptoms of COVID-19 justify its promise as a repurposed bioactive secondary metabolite against COVID-1924. The HPTLC analysis of methanol extract and aqueous extract of Ayurvedic Polyherbal Tablet Formulation confirmed 0.46% and 0.14% of quercetin respectively (Figure 6,7,8 and 9).

 

Fig. 5 - Correlation of Rf value for methanol extract of sample, methanol extract of Carica papaya, Vettivera zizannoides and standard gallic acid at 254 nm. T1- methanol extract of sample, T2-aqueous extract of sample T3-Standard Rutin, Gallic acid, Quercetin, T4-Cyperus rotundus methanol extract, T5-Mollugo cerviana methanol extract, T6-Zingiber officinalis methanol extract, T7-Andrographis paniculata methanol extract, T8-Piper nigrum methanol extract, T9-Carica papaya methanol extract, T10-Vettivera zizannoides methanol extract, T11-Tinospora cordifolia methanol extract.

 

 

Fig. 6 - Correlation of Rf value for methanol and aqueous extract of sample and standard quercetin at 254 nm

T1- Methanol extract of marketed tablet, T2-Aqueous extract of marketed tablet, T3-Standard Rutin, Gallic acid, Quercetin, T4- Methanol extract of Melia azedarach, T5-Methanol extract of Tricosanthus dioica

 

Correlation of Rf value for methanol extract of sample, methanol extract of Carica papaya, Vettivera zizannoides and standard gallic acid at 254 nm. T1- methanol extract of sample, T2-aqueous extract of sample T3-Standard Rutin, Gallic acid, Quercetin, T4- Cyperus rotundus methanol extract, T5-Mollugo cerviana methanol extract, T6-Zingiber officinalis methanol extract, T7-Andrographis paniculata methanol extract, T8-Piper nigrum methanol extract, T9-Carica papaya methanol extract, T10-Vettivera zizannoides methanol extract, T11-Tinospora cordifolia methanol extract.

 

Fig. 7 - Densitogram of a) methanol extract of sample b) aqueous extract of sample c) methanol extract of standard rutin, quercetin, gallic acid d)methanol extract of Cyperus rotundus e) methanol extract of Mollugo cerviana f) methanol extract of Zingiber officinalis g) methanol extract of  Andrographis paniculata h) methanol extract of Piper nigrum i) methanol extract of Carica papaya j) methanol extract of Vettivera zizannoides k)methanol extract of Tinospora cordifolia at 254 nm

 

Fig. 8 - Correlation of Rf value for methanol and aqueous extract of sample and standard quercetin at 254 nm

 

Fig. 9 - Densitogram of a) methanol extract of sample b) aqueous extract of sample c) methanol extract of standard rutin, quercetin and gallic acid d) methanol extract of Melia azedarach e) methanol extract of Tricosanthus dioica at 254 nm

 

Quercetin is an anti-inflammatory, antioxidant, analgesic and NLRP3 inflammasome inhibitor compound. Hence, it is used as a potential treatment for severe inflammation, which is the main life threatening conditions in patients with COVID-1925 (Table 2).

 

Table 2- Rf values for markers in tablet extracts and raw material extracts

S. No

Name of sample

Rf values of compounds in extracts of crude material andsample extract

Rf value of marker in extracts

Name of marker in extracts

Area of standard marker sample

Amount of marker present in sample

Percent of marker present in sample

1

Andrographis paniculata

0.07, 0.13, 0.15,

0.20

Rutin

1489.6

0.19 μg

0.09 %

0.20, 0.28, 0.32,

 

 

 

 

 

0.40, 0.47, 0.56,

 

 

 

 

 

0.62, 0.69, 0.76, 0.83, 0.87

0.56

Andrographolide

26053.3

9.82 μg

1.96 %

 

0.76

Gallic acid

8271.0

0.98 μg

0.49 %

 

0.83

Quercetin

11010.9

1.42 μg

0.71 %

2

Piper nigrum

0.07, 0.10, 0.15,

0.17

Rutin

539.8

0.06 μg

0.03 %

0.17, 0.23, 0.39,

 

 

 

 

 

0.44, 0.59, 0.61, 0.69, 0.75, 0.85,

0.85

Quercetin

1099.9

0.14 μg

0.07 %

0.91

0.59

Piperine

38849.9

5.67 μg

1.13 %

 

0.75

Gallic acid

27741.9

3.31 μg

1.66 %

3

Methanol extract of

0.18, 0.23, 0.31,

0.18

Rutin

574.5

0.36 μg

0.04 %

 

Ayurvedic Polyherbal

0.46, 0.54, 0.57,

0.54

Andrographolide

1427.1

1.07 μg

0.11 %

 

Tablet Formulation

0.62, 0.74, 0.80,

0.57

Piperine

6831.0

1.99 μg

0.19 %

 

 

0.85, 0.88

 

 

 

0.74

Gallic acid

616.5

0.37 μg

0.04 %

 

 

 

0.85

Quercetin

7114.2

4.6 μg

0.46 %

4

Aqueous extract of

0.07, 0.11, 0.25,

0.07

Rutin

1535.4

0.95 μg

0.04 %

 

Ayurvedic Polyherbal Tablet Formulation

0.33, 0.50, 0.54,

0.56, 0.66,

0.56

Andrographolide

4286.6

3.23 μg

0.32 %

 

 

0.79,0.85

 

Piperine

6831.0

1.99 μg

0.19 %

 

 

 

0.79

Gallic acid

2838.0

1.69 μg

0.17 %

 

 

 

0.85

Quercetin

2188.2

1.40 μg

0.14 %

5

Andrographolide

0.56

13252.2

6

Piperine

0.57

34219.6

7

Gallic acid

0.76

16725.4

8

Quercetin

0.85

15549.9

9

Rutin

0.19

16088.7

 

Table 3 - Determination of heavy metals in ayurvedic polyherbal tablet formulation

Heavy metals (mg/l)

Ayurvedic Polyherbal Tablet Formulation

WHO Permissible limit

Arsenic

0.15 ppm

3 ppm

Cadmium

Not detected (LOD: 0.05 ppm)

0.3 ppm

Lead

2.0 ppm

10 ppm

Mercury

0.20 ppm

1 ppm

 

Heavy metal analysis showed the absence of Cadmium and levels of Arsenic, Lead, and Mercury within WHO permissible limits (Table 3). Microbial contamination tests revealed no bacterial or fungal growth, confirming the formulation's safety for internal use.

Microbial contamination test results revealed the absence of bacteria and fungus in the formulations, indicating that the formulation is safe for internal use (Figure 10) (Table 4).

 

Test sample-1                                              Test sample-2                                                 Test sample-3

 

(a)

Test sample-1                                              Test sample-2                                                 Test sample-3

 

(b)

Figure 10. Photograph showing the absence of a) bacterial growth in the Ayurvedic Polyherbal Tablet Formulation b) fungal growth in the Ayurvedic Polyherbal Tablet Formulation

 

 

Table 4 - Determination of microbial contamination in the ayurvedic polyherbal tablet formulation

S. No

Parameters

Results

WHO Permissible Limits

Test sample 1

Test sample 2

Test sample 3

 1

Total bacterial count

Absent

Absent

Absent

105/g

 2

Total fungal count

Absent

Absent

Absent

103/g

 

DISCUSSION

The observed weight variation of the tablets, ranging from 0.10% to 1.73%, demonstrates excellent uniformity, well within the acceptable pharmacopoeial limit of ±5% deviation. This indicates consistent manufacturing processes with minimal variability in the weight of individual tablets. The results reflect high precision in tablet formulation and compression, suggesting effective control over raw material handling, granulation, and tablet pressing parameters.

 

The disintegration time being well within pharmacopoeial limits indicates efficient tablet formulation and suitability for rapid dissolution. The extractive values highlight a significant presence of water-soluble phytochemicals, supporting the tablet's therapeutic potential.

 

The pH of 8.20 suggests compatibility with physiological conditions, which is beneficial for patient compliance. The HPTLC findings validate the presence of andrographolide, piperine, and flavonoids, which contribute to the formulation's pharmacological effects, such as antiviral, antioxidant, and anti-inflammatory properties. For instance, andrographolide's ability to inhibit COVID-19 protease and enhance immune response and piperine's antiviral potential through RNA-binding pocket inhibition align with contemporary therapeutic needs.

 

The identification of gallic acid, rutin, and quercetin further strengthens the tablet's utility, given their antioxidative and anti-inflammatory roles. Specifically, rutin and quercetin's inhibition of viral protease enzymes underscores their promise in managing COVID-19 symptoms.

 

The absence of heavy metals and microbial contamination underscores the formulation's safety. Adherence to WHO standards ensures that the product is suitable for long-term use without adverse effects from toxic substances or microbial growth.

 

CONCLUSION

Evaluation tests such as extractive value, disintegration time, pH and the phytochemical investigation of Ayurvedic Polyherbal Tablet Formulation were found to be within the limits. HPTLC investigations of methanol and aqueous extracts of the Ayurvedic Polyherbal Tablet Formulation confirmed the presence of the active ingredients mentioned in the Label claim. Microbial tests and heavy metal analysis revealed that the formulation is safe as it does not contain any microorganisms or heavy metals. In conclusion, the ayurvedic polyherbal tablet demonstrates a robust safety profile, coupled with significant therapeutic potential, particularly in antiviral applications. Future studies could focus on further optimizing the formulation and exploring its clinical efficacy in managing viral infections.

 

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

 

ACKNOWLEDGEMENTS:

The authors were thankful to management of KMCH College of Pharmacy, Tamil Nadu, India for providing necessary facilities to carry out the study.

 

REFERENCES:

1.      Srikanth N, Ramesh Babu. Research and Development in Ayurveda –An overview. International Conference on Global scenario of Traditional system of Medicine, Ayurveda, Agriculture and Education. 2013.

2.      Uma Narayanamurthy, Mirunalini R, Subha V, Manimekalai K, Sakthibalan K, Arther Paul C, Nagarajan KM and Sabarianandh JV. Acute and Reported Dose Toxicity Study of Clevira Syrup – A Polyherbal Formulation. Biomedical and Pharmacology Journal. 2021; 14(3):1459-1467.

3.      Karthikeyan Lakshmanan, Arivukkarasu Ramasamy, Rajasekaran Aiyalu, Hari Baskar Balasubramanian, Anguraj Moulishankar. Elucidation of plausible mechanisms of kabasura kudineer for covid-19 by molecular docking against SARS COV-2. Indo American J of Pharmaceutical Sciences. 2020;7:((7):1795-1802.

4.      Esther Lydia, Sheila John, Riyazudin Mohammed and Thiyagarajan Sivapriya. Investigation on the Phytochemicals present in the Fruit peel of Carica papaya and evaluation of its Antioxidant and Antimicrobial property. Research Journal of Pharmacognosy and Phytochemistry. 2016; 8(4):217-222.

5.      Rajurkar RM, Jain RG, Bedmohta PA and Khadbadi SS. Antioxidant Activity of Phenolic Extract from Ginger (Zingiber officinale Roscoe) Rhizome. Asian Journal Research Chemistry. 2009;2(3):260-261

6.      Lydia J. and Sudarsanam D. Total phenol and total tannin content of Cyperus rotundus L. and its medicinal significance. Research Journal of Pharmacy and Technology. 2012; 5(12):1500-1502.

7.      Vijaya Kumar S, Dhirendra B, Sanghai Mallikarjuna Rao C, Shreedhara C. S. Antioxidant and antihyperlipidemic activity of Melia azedarach Linn. extracts in rats. Research Journal of Pharmacy and Technology. 2013; 6(11):1195-1199

8.      Nirupama Rani Dewangan, Avantika Agrawal, Bharti Ahirwar. Novel Therapeutic Approach of Mollugo cerviana (L.) Ser Plant: A Review. Research Journal of Pharmacy and Technology. 2022; 15(7):3280-3284.

9.      NB Ghiware and TM Nesari.Antipyretic Activity of Piper nigrum and Nyctanthes arbor-tristis in Different Dosage Forms. Research Journal of Pharmacy and Technology 2010;3(1): 157-160.

10.   Ragavee A, Tenzin Choedar, Sourabh Suman, S. Asha Devi. Evaluation of Bioactive Constituents and Antioxidant Activity of Mucilage Isolated from Tinospora cordifolia. Research J. Pharm. and Tech 2018; 11(7): 2747-2751.

11.   Wajid Azam, Vicky Rai, Rehan Jawed, Kalyani Jaywardhane, Vishal Thakur, Vishal Jain, Parwez Alam.Comparison of Antifungal and Antimicrobial activities in different parts of Carica papaya Linn.  Asian Journal of Pharmaceutical Research. 2023;13(4):213-218.

12.   Raj K. Tiwari R. Pandey S.S. Shukla, Prashant Tiwari, H. Shah. Antibacterial Activity of Aerial Part of Andrographis paniculata Research Journal of Pharmacognosy and Phytochemistry. 2014; 6(3): 122-125.

13.    Lohar DR. Protocol for testing Ayurvedic, Siddha & Unani medicines, Department of AYUSH, Ministry of Health & Family Welfare, Pharmacopoeial Laboratory for Indian medicines. 2007.

14.   Pulok K. Mukherjee. Quality control and evaluation of herbal drugs. Published by Elsevier Science. First Edition. 2019.

15.   Chaowalit Monton, Worawan Saingam, Jirapornchai Suksaeree, Apirak Sakunpak, Krisana Kraisintu. Preformulation and Physical Properties study of Fast Disintegrating Tablets from Thai Traditional Formula. International Journal of Pharmacy and Pharmaceutical Sciences. 2014; 6(4): 431-434.

16.   Pattanayak P, Mohapatra P, Jena RK and Panda SK. Standardization of Sulaharan Yoga: An Ayurvedic Tablet Formulation. Indian Journal of Pharmaceutical Sciences. 2011; 73(1): 65-70.

17.   Quality control methods for medicinal plant materials. Published by WHO. Geneva. 1998;64-73.

18.   Standard Operating Procedures. Determination of metals by Inductively Coupled Plasma methods. Published by Seras. 2006:1-42.

19.   Marimuthu M, Mohanraj, Manojkumar D, Melina D. Cruz, Rajasekaran A, Arivukkarasu R. Estimation of Andrographolide Content in Aqueous and Methanolic Extracts of Marketed Nilavembu Kudineer Siddha formulations by HPTLC Technique. Asian Journal of Pharmaceutical Analysis. 2023:13(4): 239-242.

20.   Rajasekaran A, Linda A, Arivukkarasu R. Estimation of Andrographolide Content in Aqueous Leaf Extract of Andrographis paniculata (Burm. f.) Wall. ex Nees from Different Regions of Tamilnadu and Kerala. Inventi Rapid: Pharm Analysis & Quality Assurance. 2016; 1-18.

21.   Choudhary, Prassan, Chakdar, Hillol, Singh, Dikcha, Selvaraj, Chandrabose, Singh, Sanjeev Kumar, Kumar, Sunil, Saxena, Anil Kumar: Computational studies reveal piperine, the predominant oleoresin of black pepper (Piper nigrum) as a potential inhibitor of SARS-CoV-2 (COVID-19). Current Science. 2020; 119(8): 1333-1342.

22.   Arivukkarasu R, Rajasekaran A, Sethuramani, Kattu raja, Mohamed Ali Jinna, Nandhini, Ramya M Vinothini R. Detection and estimation of well-known free radical scavengers rutin, quercetin and gallic acid in market herbal anti-inflammatory and anti- arthritis formulations by HPTLC Methods. Scholars Academic Journal of Pharmacy (SAJP). 2017; 6(5): 221-228.

23.   Nourah Mohammad Abd El-Aziz, Mohamed G. Shehata, Olfat M. Eldin Awad, Sobhy Ahmad El-Sohaimy. Inhibition of COVID-19 RNA Dependent RNA Polymersae by Natural Bioactive Compounds: Molecular Docking Analysis. Egyptian Journal of Chemistry. 2021; 64(4): 1989-2001.

24.   Pawan K. Agrawal, Chandan Agarwal, Gerald Blunden. Rutin: A potential antiviral for repurposing as a SARS-CoV-2 Main Protease inhibitor. Natural Product Communications. 2021; 1694: 1-12.

25.   Ali Saeedi-Boroujeni, Mohammad Reza, Mahmoudian Sani. Anti-inflammatory potential of Quercetin in COVID-19 treatment. Journal of Inflammation. 2021; 18(3): 1-11.

 

 

 

Received on 23.05.2025      Revised on 27.06.2025

Accepted on 29.07.2025      Published on 10.10.2025

Available online from October 18, 2025

Res. J. Pharmacognosy and Phytochem. 2025; 17(4):258-268.

DOI: 10.52711/0975-4385.2025.00042

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