IJCRR

IJCRR - 14(4), February, 2022

Pages: 56-62

Date of Publication: 15-Feb-2022

Development and Assessment of In-vitro Antioxidant Potential of Differently Flavoured Poly Herbal Green Tea Formulations

Author: Samidha M. Pawaskar, Ketan Ranade

Category: Healthcare

Abstract:Introduction: Green tea is a drink relished throughout the world and also has positive health benefits. Its antioxidant properties can be increased using different combination of herbs. Aim: The aim of the research was to develop various flavoured poly herbal green tea formulations and assess their Invitroantioxidant potential. Method: Ingredients used in the base formulation of polyherbal green tea were leaves of Camellia sinensis (L) Kuntz (Tea), Cymbopogon citratus (DC) Stapf. (Lemon grass), Rosemarinus officinalis (Linn.)(rosemary)and petals of Rosa Centifolia(Linn.) and Hibiscus rosa-sinensis (Linn.). Three variants of the above-mentioned base formulations were made by flavouring them with leaves of Ocimum tenuiflorum (Linn.) (Holy Basil or Tulsi), Mentha piperita (Linn.) (Peppermint) and roots of Zingiber officinale (Rosc.) (ginger). All the three formulations were quantitatively assayed for their total antioxidant capacity, total phenolic capacity, reducing power and scavenging activities. Results: Our study indicated that the total antioxidant capacity, total phenolic content, reducing power and hydroxyl radical scavenging activity was found to be highest in Tulsi flavoured poly herbal green tea and lowest in peppermint flavoured tea. Total flavonoid content was found to be highest in Tulsi and lowest in ginger-flavoured poly herbal green tea. Nitric oxide scavenging activity was found more in peppermint poly herbal green tea than in the other two formulations until a certain concentration (7.5 mg/ml) where the scavenging activity was almost equal for all the three formulations. Above 7.5mg/ml concentration the nitric oxide scavenging activity was observed to be more in ginger flavoured poly herbal green tea followed by tulsi and then peppermint flavoured poly herbal green tea. Conclusion: Thus the natural poly herbal flavoured green tea formulations developed were proved to have good antioxidant potential.

Keywords: Camellia sinensis (L) Kuntz, Cymbopogon citratus (DC) Stapf., Rosemarinus officinalis (Linn.), Rosa Centifolia (Linn.), Hibiscus rosa-sinensis (Linn.), Ocimum tenuiflorum (Linn.), Zingiber officinale (Rosc.), Mentha piperita (Linn.), Total antioxidant capacity, Total phenolic content, Reducing power and scavenging activities

Full Text:

Introduction:

Antioxidants are compounds that protect cells against the damaging effects of free radicals and reactive oxygen species. Any imbalance between the antioxidants and reactive oxygen species results in oxidative stress. 1 In normal physiological conditions also free radicals and reactive oxygen species (ROS) are generated during metabolic processes leading to oxidative stress. However if they are not eliminated then the conditions could lead to complications.² Oxidative stress can cause oxidative damage to large biomolecules such as proteins, DNA and lipids, resulting in an increased risk of cancer, cardiovascular disease, neural disease, Alzheimer’s disease, Parkinson’s disease, ulcerative disease, aging and atherosclerosis.^2,3Antioxidant compounds such as carotenes, phenols, phytates, etc. present in food are important for neutralizing free radicals, thus preventing ageing process and also reducing the risk from various related diseases.^4,5

Synthetic antioxidants could reduce oxidative damages but could lead to carcinogenic negative effects on liver and lungs. Thus, studies of antioxidants are focused on naturally occurring substances, especially on plant phytochemicals.

Thus the present study aimed to formulate a naturally flavoured poly herbal green tea, using leaves of Camellia sinensis (L) Kuntz(Tea), Cymbopogon citratus (DC) Stapf (Lemon grass), Rosemarinus officinalis (Linn.) (rosemary), and petals of  Rosa Centifolia(Linn.) and Hibiscus rosa-sinensis (Linn.)and to assess the in-vitro antioxidant capacity for these formulations. Three variants of the above-mentioned base formulation were made by flavouring them with leaves of Ocimum tenuiflorum (Linn.) (Holy Basil or Tulsi), Mentha piperita (Linn.) (Peppermint) and roots of Zingiber officinale (Rosc.) (ginger).

Tea (Camellia sinensis (L) Kuntz) is the most widely consumed beverage in the world⁶ and green tea is known to have several groups of polyphenols.^1,7 Thus combinations made of such natural drinks would be useful to deliver health benefits to the masses.

Materials and Methods:

Preparation of polyherbal flavoured green tea formulations:

Ingredients used in the preparation of base formulation of polyherbal green tea were Camellia sinensis (L) Kuntzleaves, Cymbopogon citratus (DC) Stapfleaves, Rosemarinus officinalis (Linn.) leaves, Rosa Centifolia (Linn.) petals and Hibiscus rosa-sinensis (Linn.) petals. Three variants of this base formulation was made by flavouring them with the leaves of Ocimum tenuiflorum (Linn.) and Mentha piperita (Linn.)and roots of Zingiber officinale (Rosc.). All the ingredients were procured from local market, washed thoroughly with water and were oven dried.

Extract Preparation:

All the samples were prepared by adding hot water to tea leaves (10 mg/ml), then the mixture was kept at room temperature for 10 minutes before starting the experiments.

The assays for total antioxidant capacity, total phenol content, total flavonoid Content (flavones and flavonols), reducing power, scavenging ability for hydroxyl radicals, nitric oxide radical scavenging, were performed using known and standardized methods.^{8 – 14}     

Statistical Analysis:

All data were expressed as the means ± SD of three independent experiments carried out in triplicate.

Results:

The results of the antioxidant potential of differently flavoured polyherbal tea formulations viz., total antioxidant capacity, total phenolic content and total flavonoid content has been represented in table 2, table 3 and table 4, respectively. Results of reducing power assay has been represented in table 5.  NO scavenging assay and OH scavenging assay has been represented in table 6 and graph 1, graph 2, and graph 3 respectively.

Discussion:

Polyherbal flavoured green tea formulations:

Dried ingredients were ground and mixed in exact quantity to make the base formulations of polyherbal green tea. Three different flavours were created. The exact composition is as shown in (Table 1). Cymbopogon citratus (DC) Stapf.(Lemon grass)has various phytoconstituents such as flavonoids and phenolic compounds, which consist of luteolin, isoorientin 2'-O-rhamnoside, quercetin, kaempferol and apigenin reported to have antioxidant effects.¹⁵Rosemarinus officinalis (Linn.) has been known as a versatile, aromatic herb known for its powerful antioxidant activity, antibacterial and antimutagenic properties as it contains high concentration of phenolic substances obtained from the leaves of the plant. The antioxidant activity of rosemary was found mainly due to phenolic compounds responsible for rosemary antioxidant activity like phenolic di-terpenes such as carnosol, carnosic acid, rosmanol, epirosmanol and isorosmanol.¹⁶ The rose flowers and leaf extracts of Hibiscus rosa-sinensis (Linn.)were also found to have high antioxidant activities.¹⁷

 In the present study, three variants of the above-mentioned base formulation were made by flavouring them with leaves of Ocimum tenuiflorum(Linn.) (Holy Basil or Tulsi), Mentha piperita (Linn.) (Peppermint) and roots of Zingiber officinale (Rosc.) (ginger).

Total antioxidant activity:

The total antioxidant activities of all three flavoured natural polyherbal green tea formulations were examined and expressed as gallic acid equivalents (Table 2)

The total antioxidant capacity of Ocimum tenuiflorum(Linn.), Zingiber officinale (Rosc.)and Mentha piperita (Linn.)flavoured poly herbal green tea in our study is reported in Table 2. We found that the total antioxidant capacity of Ocimum tenuiflorum(Linn.) flavoured poly herbal green tea was 30% more than Zingiber officinale (Rosc.)flavoured poly herbal green tea.(Table 2). Pandey et al, 2014 reported that Ocimum tenuiflorumof family Labiateae is a good source of linalool, eugenol, methyl charicol and cineole. Presence of eugenol contributes to its antioxidant property and is also thought to be responsible for inhibition of lipid peroxidation.⁶This could be the reason for very high antioxidant activity of Ocimum tenuiflorum(Linn.) found inthe present study.

Total Phenolic Content and Flavanoid content:

Total phenolic content of all three flavoured natural polyherbal green tea formulations was examined and expressed gallic acid equivalents. Standard graph of gallic acid for total phenolic content is represented in Table 3 and Total flavonoid content was examined and expressed as mg of Quercetin equivalents (QE)/g of the plant material. (standard graph of quercetin for total flavanoid content is represented in (Table 4)

The total phenolic content of Ocimum tenuiflorum (Linn.), Zingiber officinale( Rosc.) and Mentha piperita (Linn.)flavoured poly herbal green tea was found to be 482.4  +  22 , of 360.0  +  24  and 280.0 +18mgs of gallic acid equivalents (GAE)/g of the plant material respectively.  It was found that phenolic content of Ocimum tenuiflorum(Linn.) flavoured tea was 34% higher than the Zingiber officinale (Rosc.) flavoured poly herbal green tea. Our results are nearly similar to the results of Somia Lessed et al who reported total phenolic content of 394.66+  22.67 mgs of gallic acid equivalents (GAE)/g of the plant material.¹⁸ Kaurinovic B et al. also reported significant amount of total phenolic content in the water extracts of Ocimum basilicum.¹⁹

The total flavonoid content of Ocimum tenuiflorum(Linn.), Zingiber officinale (Rosc.)and Mentha piperita (Linn.) flavoured poly herbal green tea was found to be 32.60   +  1.60, 20.69    +  0.60 and 31.661   +0.98mgs of Quercetin equivalents (QE)/g of the plant material. One such study reported the total flavonoid content of 5.04 +  0.59 mgs of Quercetin equivalents (QE)/g of the plant material.¹⁸ However the medium used for the extraction by them was chloroform, whereas we had used plain water for the said extraction. Mckay et al. reported that the Mentha piperita (Linn.)leaves contain phenolic constituents including rosmarinic acid and several flavonoids, primarily eriocitrin, luteolin and hesperidin and show strong antioxidant potential.²⁰

Reducing Power Assay:

The results in this study showed that the reducing power was observed to be increasing with the concentration of the extract (Figure1) (Table5). The reducing capacity was found to be increasing in the order Mentha piperita (Linn.) flavoured poly herbal green tea <Zingiber officinale (Rosc.) flavoured poly herbal green tea <Ocimum tenuiflorum (Linn.) flavoured poly herbal green tea < Ascorbic oxide standard. Significant association of Reducing power and the antioxidant activity has also been documented.²¹ Compounds with reducing power indicate that the electron-donating property can reduce the oxidized intermediates of lipid peroxidation processes and thus they can act as primary and secondary antioxidants.²²

Nitric oxide radical scavenging (NO) assay:

Nitric oxide scavenging activity was found to be more in Mentha piperita (Linn.)flavoured poly herbal green tea than in the other two formulations until a certain concentration (30 mg/ml) where the scavenging activity was almost equal for all three formulations (Figure 2) (Table 6). Above 30 mg/ml concentration the nitric oxide scavenging activity was observed to be more in Zingiber officinale (Rosc.)flavoured poly herbal green tea followed by Ocimum tenuiflorum (Linn.) flavoured poly herbal green tea and then the Mentha piperita (Linn.) flavoured poly herbal green tea. The nitric oxide scavenging activity of the standard ascorbic oxide was found to be more compared to all three formulations, at any given concentrations.

Hydroxyl radical scavenging (HO) assay:

Hydroxyl radical is one of the potent reactive oxygen species present in the biological system. These ions damage cells by interacting with the phospholipids of the cell membrane.²³ Hydroxyl radical scavenging activity was observed to be decreasing in the order standard ascorbic acid >Ocimum tenuiflorum(Linn.) flavoured poly herbal green tea >Zingiber officinale (Rosc.)flavoured poly herbal green tea >Mentha piperita (Linn.) flavoured poly herbal green tea. It increased as the concertation of standard and the extracts increased (Figure 3) (Table 6).

Conclusion:

Natural polyherbal green tea formulations in three flavours were developed. All three flavours were studied for their total antioxidant capacity, total phenolic content and scavenging activities. All three flavours of natural polyherbal green tea formulation developed were proved to have good antioxidant potential and can have potential application as therapeutics.

Acknowledgement: Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.

Source of funding: University of Mumbai

Conflicts of interest: The authors declare no competing interests.

Authors’ Contribution: All the authors of this paper equally contributed towards the experimental work as well as the writing of this paper.

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