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Article

Evaluating the Phytochemical Composition and Antioxidant Activity of Leaves of Different Rose Varieties

by
Shivani Sharma
1,
Lipakshi Awasthi
1 and
Poonam Kumari
1,2,*
1
Division of Agrotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur 176 061, India
2
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201 002, India
*
Author to whom correspondence should be addressed.
Int. J. Plant Biol. 2023, 14(4), 1051-1063; https://doi.org/10.3390/ijpb14040076
Submission received: 30 August 2023 / Revised: 30 September 2023 / Accepted: 3 October 2023 / Published: 16 November 2023
(This article belongs to the Section Plant Physiology)
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Abstract

:
Rose is a commercially significant floricultural crop that has been used for various industrial as well as decoration purposes. Along with the beautification of rose flowers, their leaves are enriched with different biologically active compounds having various therapeutic uses. The current study was performed on the phytochemical and antioxidant activity of aqueous extracts of rose leaves. In our study, we found there are consequential variations observed in all the parameters, viz., total chlorophyll, carotenoids, total anthocyanin, total phenol, flavonoids, and antioxidant activity, among all the varieties. Among the ten varieties, Thelma Barlow revealed the maximum phenolic content (35.19 mg/g FW), flavonoids content (15.97 mg/g FW), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity of IC50 value (206.86 ± 0.49 µg/mL), and 2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) (301.62 ± 2.31 µg/mL). The variety Grand Amore presented the highest values for total chlorophyll (1.41 mg/g FW) and carotenoids (36.29 mg/g FW) content compared to other varieties. Also, a comparative correlation was studied amongst the phytochemicals such as anthocyanin content, total phenolic content, flavonoids, and antioxidant activities. Amongst the different rose varieties, Grand Amore and Thelma Barlow contain higher antioxidant potential, owing to their greater phytochemical activity. From our findings, we collectively concluded that fresh rose leaves contain potentially higher phenolic and flavonoid content, i.e., are responsible for higher antioxidant activity, which can be utilized for various pharmacological as well as food industries.

1. Introduction

Rose is an important ornamental plant grown worldwide and has been called the queen of flowers. It is a perennial woody plant belonging to the genus Rosa. Currently, there are 200 species, and more than 30,000 cultivars are known to be present throughout the Asian region, Middle East region, North America, and Europe [1,2,3]. Rose species easily hybridize from all over the world, giving rise to the various garden rose cultivars. Rosa also has 435 taxa, 308 species, 13 subspecies, 28 variations, 17 forms, and 71 natural hybrids. Because of its essential oils and volatile compounds, this is one of the largest and main aromatic and medicinal genera in this family, significant in traditional medicine, pharmacology, and commercial value [4]. It is used to cure several diseases like coughs, eczema, itching, and colds, and therefore it is considered a medicinal plant in Turkey. Moreover, previous studies on rose secondary metabolites confirmed that the leaves are a promising source of bioactive compounds [5]. Rose leaves have many medicinal properties, so they can be called medicinal herbs, which are very useful as a blood purifier. In ancient times, the plant leaves were effective in treating various health issues like intestinal ulcers, diarrhea, etc. [1]. In Ayurveda, whole plants have been used for useful purposes. The petals, leaves, hips, stems, and roots of a rose plant contain various phytochemical components and nutrients in the form of vitamins and minerals. Its leaves contain several compounds like phenols, flavonoids, tannins, fatty acid derivatives, enzymes, vitamins, minerals, folic acid, and terpenoids [4]. From previous rose studies, it is well known that flavonoids and other phenols found in plants are good sources of human diet and have a wide and extensive array of biological properties, such as anti-allergenic, anti-inflammatory, anti-microbial, anti-thrombotic, cardio-protective, and vasodilatory effects [5]. For instance, the liquid prepared from the fresh leaves is mostly given in doses of 5–30 drops. In small doses, it will calm a person but must be given with care; it may cause an upset stomach, drowsiness, and dizziness. Consequently, there is an increasing interest in edible plants with antioxidants and phytochemicals as potential therapeutic agents [6,7]. Numerous studies have found that chlorophyll content in the leaf is a predominant physiological as well as a morphological parameter that can serve nutritional status, plant stress, or senescence. Also, carotenoids have a vital role in the proper functioning of biochemical processes in animals and humans, including eyesight (pro-vitamin A), growth, and reproduction [8]. In a study, it was found that roses contain some compounds that may contribute to relaxation and anxiety reduction and are consumed as tea. A well-known species, Rosa indica, exhibits antimicrobial properties and is used to cure several diseases like diarrhea, asthma, leukoderma, and oral irritation. As described in the literature, the color value is one of the most important quality parameters and is a visual miracle that attracts customers. Therefore, the color of the food source is the first criterion followed by users to evaluate and is a crucial step for product acceptance [9]. Besides this, the Rosaceae family is of great significance because of their use in several food preparations and perfumery. Commercially, rose producers do not learn much about plant leaves and their therapeutic uses. Compared to fruits and flowers, leaves are not explored for much research [10]. In selected rose varieties, there is richness in secondary metabolites (anthocyanin, chlorophyll, and carotenoids) as well as antioxidant compounds. Therefore, the present investigation aimed to estimate the differences in pigment contents (chlorophylls and carotenoids), secondary metabolites (phenols and flavonoids), color measurements, and antioxidant activity of the leaves.

2. Materials and Methods

2.1. Sample Collection

The present study was carried out on the ten Rosa x hybrida varieties, viz., Thelma Barlow, Le Rouge et le Noir, Majestic Burgundy, Grand Amore, Swamy, Silver Shadow, Louis Estes, Dr. N.C. Sen, Whippet, and Ma Normandie (Table 1), collected from the Rose Garden of CSIR- Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh (Figure 1). Young rose leaves (4–5 leaves) were harvested in March 2022 and are used for further biochemical analysis.

2.2. Total Chlorophyll and Carotenoid Content

Two hundred milligrams of fresh rose leaves were weighed and ground in 10 mL of 80% acetone with the help of a pestle mortar; this was centrifuged at 10,000 rpm for 15 min. Over time, the supernatant was collected, and 10 mL of 80% acetone was added again. The extraction mixture was vortexed properly. Again, the supernatant was collected to a final volume of 20 mL. After this, the absorbance was recorded at different wavelengths of 663 nm, 645 nm, and 480 nm, respectively. The total chlorophyll and carotenoid content in leaf extract samples was estimated in mg/g [11,12].

2.3. Total Phenolic Estimation

The total phenol present in the leaf sample was estimated by using the colorimetric method (Folin–Ciocalteu) which relies on a redox reaction (oxidation and reduction). Different concentrations of trihydoxybenzoic acid (GAE) in methanol (10, 25, 50, 75, and 100 μg/mL) were used for phenol determination. In each concentration, 1 mL of GA was added to 5 mL of FC mixture (10%) and 4 mL of Na2CO3 (7%) to a final volume of 10 mL. The bluish-colored mixture was mixed thoroughly and kept for 30 min in a boiling water bath at 40 °C. The reading was measured at wavelength 760 nm alongside a methanol blank in a spectrophotometer. The experiment was carried out in three biological replicates. For the test sample, 1 mL of methanol extract of rose leaf dilutions was pipetted instead of gallic acid solutions. The total phenol content in rose leaf samples was estimated as mg/g [13].

2.4. Total Flavonoids Estimation

For the determination of flavonoids, quercetin is used as a standard. Twenty milligrams of quercetin in 100 mL of methanol (95%) was used for standard solution preparation. For working standard solutions, 30, 60, 90, 120, 150, and 180 micrograms per milliliter were prepared by diluting the standard stock. For total flavonoids estimation, 100 μL of sample extracts was incubated with 1.5 mL of 95% methanol, and 0.1 mL of aluminum chloride (10%), 0.1 mL of 1 M potassium acetate, and 2.8 mL of distilled water was added to each test tube. The reaction mixtures were thoroughly mixed and incubated for 30 min at room temperature. The absorbance was measured at 415 nm wavelength using a spectrophotometer. The blank was prepared by substituting the same amount of aluminum chloride (10%) with distilled water. The total flavonoid content in rose leaf sample extracts was expressed in mg/g [14].

2.5. Determination of Antioxidant Activity

2.5.1. DPPH (2,2-diphenyl-1-picrylhydrazyl) Free Radical Scavenging Assay

The antioxidant activity (DPPH) in leaf sample extracts was quantified using a method defined by [15]. 0.1 mL of ethanol extract was added to 3.9 mL of 0.0025 MDPPH (2,2-diphenyl-1-picrylhydrazyl). The sample mixture was thoroughly mixed and kept for 30 min without light at room temperature. The absorbance of the samples was measured using a spectrophotometer at wavelength 517 nm against blank, i.e., ethanol. The IC50 values of the standard and sample extracts were calculated using the following formula:
Percent inhibition (%) = [(Ao − Ae)/Ao] × 100
          (Ao = absorbance without extract; Ae = absorbance with extract).

2.5.2. ABTS (2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid)) Assay

The free radical scavenging activity of leaf sample extracts was examined by following the protocol described by [16]. The dilution of Trolox [mg/100 mL] was used as a positive control. The absorbance of sample extracts as well as the standard was performed by using the spectrometer at a wavelength of 734 nm.

2.6. Color Value

For color measurement, rose leaves were estimated by using a color spectrometer as represented in terms of “L” (lightness), “a” (redness), and “b” (blueness) values. Before the sample measurement, the spectrophotometer was calibrated using a white calibration plate. The average color values (L*, a*, b*) of color parameters were calculated for each sample. In the CIE L* a* b* color system, L* represents the range of dark-bright color from 0 to 100 (0 = black, 100 = white), whereas the* represents green to red color ranging from –60 to 60, respectively. Similarly, b* denoted the color range from blue to yellow (–60 to 60).

2.7. Statistical Analysis

All the experiments were executed thrice in biological replicates to minimize experimental error. The data were statistically analyzed by analysis of variance (ANOVA) at p-value < 0.05 using CRD design (WASP-Agri stat package 2.0).

3. Results and Discussion

3.1. Total Chlorophyll Content

Chlorophylls are Mg2+-holding tetrapyrrole molecules that contribute green color to plants. These are the main components of the photosynthetic system required for light absorption and energy transduction. Leaves and stems contain a considerable amount of chlorophyll and are responsible for photosynthesis. The chlorophyll content of plants diminishes as they mature and accumulate other pigments, including anthocyanin, carotenoids, and betalains [17,18,19]. A study shows that the regulation of Chl degradation is particularly mediated by the plant hormones signaled by ethylene, abscisic acid (ABA), and light [20].
Chlorophyll degradation prevents the production of ROS (reactive oxygen species), i.e., singlet oxygen, when protein is degraded. Chlorophyll cleavage began with the reduction of chlorophyll “a”, and “b” and resulted in the colorless linear tetrapyrrole [21]. In our case, we observed that chlorophyll “a” content in leaves of different rose varieties varied from 0.51 to 0.80 mg/g (Table 2). The maximum value noticed in the variety Grand Amore (0.80 mg/g) was found statistically significant to Le rouge et le Noir (0.79 mg/g). However, the rose variety Thelma Barlow (0.51 mg/g) exhibited a minimum value for chlorophyll “a”. Similarly, the mean value of chlorophyll “b” content and the range of variation were found in all the varieties, varying from 0.17 to 0.74 mg/g. The maximum value for chlorophyll “b” was found in the variety Grand Amore (0.74 mg/g), and the minimum value was noticed in the varieties Thelma Barlow and Silver Shadow (0.17 mg/g), respectively. The total chlorophyll revealed a significant variation of 0.74 to 1.41 mg/g in rose varieties (Table 2). Rose variety Grand Amore (1.41 mg/g) possesses the maximum total chlorophyll content. However, the minimum value for total chlorophyll was shown by Thelma Barlow (0.74 mg/g). A study by [8] reported total chlorophyll 2.9 mg/L, chlorophyll a 1.8 mg/L, and chlorophyll b 1.1 mg/L in fresh rose leaves. In another study, rose leaves showed chlorophyll “a”, chlorophyll “b,” and total chlorophyll to be 0.96 mg/g, 0.37 mg/g, and 1.34 mg/g, respectively [22]. Whereas another researcher [4] reported 0.35 mg/g chlorophyll “a”, 0.48 mg/g chlorophyll “b”, and 0.83 mg/g total chlorophyll content in fresh roseleaves. A similar finding was observed by [23] in fresh rose leaves, which is similar to our observations.

3.2. Total Carotenoid Content

Carotenoids are hydrogen- and carbon-containing compounds that belong to the tetraterpenoids (C40) class. Photosynthetic accessory pigments are the main components of plants that contribute color to various parts, mainly flowers, leaves, and fruits. They are also known as a nutritious component that is essential for human metabolism-regulating functions [24]. In our investigation, the total carotenoid content in rose varieties was evaluated, and pertinent data provided information on variation in total carotenoid (Table 3). Total carotenoids ranged from 17.76 mg/g to 36.29 mg/g. The value of total carotenoid was highest at 36.29 mg/g in the variety Grand Amore, while it was the lowest at 17.76 mg/g in the variety Thelma Barlow. A previous study by [25] observed 26.87 mg/g carotenoid content in fresh rose leaves, whereas [8] determined carotenes and xanthophylls at 57.4 mg/L in the rose leaf extract. However, in another study, the methanolic extracts of R. canina showed carotenoid content ranging from 19.76 mg/g to 25.79 mg/g in fresh leaves [4]. In the plant’s antioxidant defense system, carotene is one of the key components and is prone to oxidative damage. In extreme abiotic stress conditions, beta-carotene may be quickly degraded and no longer persist in the protection against oxidative damage [26]. In eggplants, carotenoid levels were found to increase with aging, whereas drought conditions show decreased levels of carotenoids with the excessive production of reactive oxygen species in the thylakoids [27,28].

3.3. Total Anthocyanin Content

Anthocyanins are water-soluble pigments, primarily contributing blue, purple, red, and orange color in plants. The interest in anthocyanin pigments has been raised not only in their use in food but also as beverage condiments to obtain an attractive natural red color [29]. In our study, a significant variation was observed in total anthocyanin content in rose leaves, showing variations ranging from 35.40 mg/100 g to 128.85 mg/100 g (Table 3). The maximum total anthocyanin was observed in the variety Thelma Barlow (128.85 mg/100 g), whereas the minimum value was found in the variety Grand Amore (35.40 mg/100 g). In a previous finding on fresh rose leaves, methanolic extract displayed a total anthocyanin content of 12.2 mg/100 g [5], which is similar to our results. Our findings are also similar to the results by [30], which describe total anthocyanin content ranging from 27.22 mg /100 g to 170.36 mg/100 g in fresh roseleaves. Quintinia serrata A. Cunn. shows that anthocyanin was found to be most abundant in the older leaves under canopies. They also suggested that anthocyanin is associated with photosynthesis and protects shade-adapted chloroplasts from high-intensity sunlight [31].

3.4. Total Phenol Content

Phenols is one of the main groups of secondary plant metabolites. It is chemically different from the other group of molecules that play a diverse role in plants, including a healthy defensive mechanism against various types of predators. On the other hand, plants with high phenolic content are not considered consumer-friendly because they result in poor nutritional quality, decreased digestibility, and decreased food consumption. Thus, their estimation in plant tissue is a main parameter. A significant variance was observed in phenolic content (Figure 2) in the leaves of different rose varieties. In our experiment, the data showed variations between 15.76 mg/g and 35.19 mg/g. Rose variety Thelma Barlow had a higher value (35.19 mg/g), and a lower value was observed in variety Grand Amore (15.76 mg/g). A previous study by [32] showed a total phenolic content of 17.49 mg GAE/g in the leaves of R. sempervirens, whereas [30] observed the maximum phenolic content of 73.38 mg GAE/100 g among the different rose varieties. Our results are homologous to the observation in [33], which found a total phenol of 17.20 mg GAE/g in the methanolic extract of rose leaves. Another researcher studied the biochemical properties of some wild and cultivated rose leaves and found 33.63 mg GAE/g total phenol in rose leaves [10]. Similarly, findings reported by [5] total polyphenols of 36.1 mg CAE/g to 46.8 mg CAE/g in the methanolic extract of fresh rose leaves.

3.5. Total Flavonoid Content

In phenolics, flavonoids are one of the naturally distributed secondary metabolites in plants. The flavonoids exhibit an anti-oxidative effect, which results in the inhibition of lipid peroxidation to scavenge free radicals and active oxygen, chelate iron ions, and inactivate lipoxygenase [34]. It has been found that the presence of higher phenols and flavonoids in plants contributes to their antioxidant potential. In our study, we found variation in the range from 4.10 mg/g to 15.97 mg/g. Rose variety Thelma Barlow leaves had a higher value (15.97 mg/g), and lower values were observed in Grand Amore leaves (4.10 mg/g) (Figure 2). In previous findings, it was found that flavonoid is the main quencher of singlet oxygen in the presence of very strong visible light. Due to the short dispersion distance of singlet oxygen, only flavonols occur in the cytoplasm, and they may protect cells by interacting with the destructive molecules. Chlorophyll breakdown may improve flavonol biosynthesis due to the leaves having less chlorophyll being more efficiently lit than densely colored leaves [35,36]. Our findings are similar to those of various researchers [32], who observed total flavonoids of 9.85 mg CE/g in rose leaves. The total flavonoid content in the methanolic extract of rose leaves was determined by [33], i.e., 10.21 mg QE/g. Similarly, another study reported 3.91 mg QAE/g to 8.04 mg QAE/g total flavonoid content in rose leaves [7]. A similar finding has been observed in [4], which reported 13.1 mg QE/g to 14.7 mg QE/g total flavonoid content in fresh rose leaves.

3.6. Antioxidant Activity

3.6.1. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Free Radical Scavenging Assay

The antioxidant activity of the plants can be estimated by multiple assays holding different modes of action, including hydrogen atom transfer (HAT), single electron transfer (ET), reducing power, metal chelation, etc. DPPH free radical scavenging is the most commonly used approach and is first adopted to determine the antioxidant activity. It generally relies on the ET-based method, where HAT acts as a minor reaction pathway [37]. The DPPH method is denoted by the IC50 value, which represents the inhibitory concentration of the antioxidant to decrease the initial concentration of DPPH by 50%. Although plant polyphenols tend to have the hydrogen-donating characteristic of their hydroxyl groups, they therefore function as reducing agents and antioxidants [38]. Xanthine oxidase is one of the flavoproteins that can catalyze the oxidation of hypoxanthine to xanthine and produce acid urate and superoxides. The leaf extract showed significant xanthine oxidase inhibition and DPPH free radical scavenging activity [39,40]. The variability of antioxidant activity was evaluated and shows a range from 111.30 ± 1.09 to 206.86 ± 0.49 (IC50 value) (Table 4). The highest value was reported in the variety Thelma Barlow (206.86 ± 0.49 µg/mL), and the lowest value was found in the variety Grand Amore (111.30 ± 1.09 µg/mL). Our study is homologous to the findings of [4], which reported IC50 values ranging from 140.7 to 266.9 µg/mL in fresh rose leaves. Another study examined the antioxidant activity of rose leaves by the DPPH method and reported an IC50 value of 56.05 µg/mL [32]. Similarly, Ref. [7] reported DPPH activity at 50.26 µg/mL in rose leaves, whereas [41] studied the DPPH free radical scavenging activity in an extract of 17 rose varieties with IC50 values ranging from 83.4 to 95.7µg/mL.

3.6.2. ABTS•+ (2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonic acid))

This assay is used to determine the ability of antioxidants to scavenge the blue-green chromophore, i.e., ABTS•+ (2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid)). It shows the highest absorption at a wavelength of 734 nm, which declines with the intensity of antioxidants. However, in the presence of powerful oxidizing agents, ABTS•+ can be generated. With the decreased intensity of antioxidants, the absorption at the wavelength of 734 nm increases. In our results, we found the ABTS values among rose varieties ranging from 177.58 ± 1.95 µg/mL to 301.62 ± 2.31 µg/mL in fresh leaves. The rose variety Thelma Barlow (301.62 µg/mL) possesses a maximum IC50 value, whereas the least IC50 value was determined in the variety Grand Amore (177.58 µg/mL). Our results are closely related to the findings of [32] reported IC50 values for the ABTS scavenging activity in rose leaves (201.5 µg/mL).

3.7. Colour Value

The CIE L*, a*, and b* values are currently one of the most popular color indicators for determining object color. The visibility of color in roses is influenced by various factors, including genetic, agronomic, and environmental, that might result in a specific type of flowering. The characteristics of plants and their genome, which determine growth and development, are the fundamental components that affect flower colors [42].
The colors of the flower, as well as the leaves in the plant, are controlled by various transcription factors and genetic pathways. The green color of the plant leaves is mainly due to the presence of high concentrations of chlorophyll. Carotenoids are one of the important accessory pigments imparting various colors to plants, such as orange, yellow, and red [43]. Besides this, anthocyanins are water-soluble pigments and are distributed in higher plants, with colors ranging from red to blue [44]. In general, variegated leaves contain green, white, and yellow sectors, which help study the biogenesis of chloroplasts and their color changes [45]. Moreover, they also show different characteristics, such as the structure of chloroplasts, the functioning of Photosystem I (PSI) and Photosystem II (PSII), chlorophyll biosynthesis, and how breakdown may result in leaf color changes [46]. A previous study showed that the biosynthesis of chloroplasts is altered by genes related to the chloroplast. The chloroplast biosynthesis mainly involves the hemA gene, which encodes glutamyl-tRNA reductase (GluTR), which is a key regulator of 5-aminolevulinic acid (5-ALA) [47,48].
In our study, we observed a significant difference in L*, a*, and b* values for both leaf surfaces, i.e., the adaxial and abaxial (Figure 3 and Figure 4). On adaxial surfaces, the variation in color value in different rose varieties was found to range from 38.13 to 14.67 for L*, 13.70 to 1.45 for a*, and 8.89 to 0.97 for b*, respectively. Rose variety Dr. N.C. Sen showed the highest value for L* (38.13), followed by Majestic Burgundy (35.54). Rose variety Swamy observed the lowest value for L* (14.67). However, the highest value for a* was observed in the variety Majestic Burgundy (13.70), which was statistically at par with Silver Shadow (12.39), and the lowest value was observed in the variety Grand Amore (1.45), followed by Dr. N.C. Sen (2.83), respectively. The variety rose Louis Estes (8.89) recorded the highest value for b*, followed by Grand Amore (8.64). Rose variety Le Rouge et le Noir exhibited the lowest b* value (0.97), which was statistically at par with Dr. N.C. Sen (2.69), Ma Normandie (2.97), Majestic Burgundy (2.99), Whippet (3.16), and variety Swamy (3.27).
We observed the leaf abaxial surface and found that the L*, a*, and b* values ranged from 36.51 to 25.27, 13.87 to 3.36, and 13.54 to 1.33, respectively (Figure 4). The highest value for L* was observed in the variety Majestic Burgundy (36.51), which was statistically on par with Ma Normandie (34.18) and Grand Amore (33.79). The lowest value was found in the rose variety Le Rouge et le Noir (25.27), followed by Thelma Barlow (27.30). Rose variety Louis Estes exhibited a higher value for a* (13.87), followed by Thelma Barlow (13.84) and Dr. N.C. Sen (13.81), respectively. The lower value for a* was recorded by the Majestic Burgundy variety (3.36), which was statistically at par with Grand Amore (7.86). However, the rose variety Silver Shadow showed the highest value for b* (13.54), and the rose Swamy showed the lowest value (1.33). Previously, similar results were observed for the color value [49,50,51].

3.8. Correlation between TAC, TPC, TFC, and Antioxidant Activities

A significant, positive correlation was found among the total anthocyanin, total phenol, total flavonoid, and antioxidant activity in rose leaves (Table 5). The correlation between these parameters shows that the high content of anthocyanin, phenolics, and flavonoids comprise an effective index for antioxidant activity in different rose varieties. Our results are in agreement with [4], which stated a strong correlation coefficient between the DPPH radical scavenging activity and the phenolic and flavonoid content in rose leaves. The research found phenolic and flavonoid content was significantly correlated with the DPPH IC50 value (r2 = 0.621 and 0.722) and ABTS IC50 value (r2 = 0.636 and 0.733) [32]. We also found that the ABTS IC50 value was highly correlated with that of DPPH (r2 = 0.998), and phenolic contents showed a good correlation with flavonoids (r2 = 0.990). In addition, we found that the phenolic compounds increased with increasing the polarity of the solvent. Therefore, the antioxidant activity of leaf extracts in the current study was directly proportional to the polyphenols.

4. Conclusions

Our present study suggests that rose leaves are a tremendous source of biologically active components with antioxidant properties. In summarizing the findings of the current study, we found differences in phytochemicals and antioxidant activity among ten rose varieties, and such differences were found to be due to genetic derivation as plants were grown using the same cultivation techniques and under the same climatic conditions. Furthermore, we observed that total flavonoid concentrations increased in proportion to total anthocyanin concentrations and that higher total phenolic concentrations resulted in higher total antioxidant activity. Anthocyanin, a natural edible pigment found in rose leaves and flower petals, is non-toxic, has high antioxidant activity, is high in phenolic compounds, and is believed to play a significant role as an antioxidant. It is also believed to have potential applications in health foods and functional cosmetics. However, the use of rose leaves in pharmaceutical industries is somewhat low in comparison to their potential due to their difficulty of storage, disease-free plant leaves, and difficulty in handling due to their softness. Therefore, we suggest that to overcome these limitations and increase the utilization of rose leaves, additional future studies on cultivation methods, storability, and production and distribution systems, as well as physiological studies on their effects on the human body, are needed.

Author Contributions

Conceptualization, P.K.; Data curation, S.S.; Formal analysis, S.S.; Investigation, P.K.; Methodology, S.S.; Data compilation, L.A.; Project administration, P.K.; Supervision, P.K.; Writing—original draft, S.S and L.A. All authors have read and agreed to the published version of the manuscript.

Funding

The study was financially supported by the Council of Scientific and Industrial Research (CSIR), grant no. HCP-0037 under the Floriculture Mission. This manuscript represents CSIR-IHBT communication number: 5429.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data are contained within the article.

Acknowledgments

Authors acknowledge the Director, CSIR-Institute of Himalayan Bioresource Technology, Palampur for supporting and providing all the necessary facilities.

Conflicts of Interest

The authors declare no conflict of interest.

Abbreviations

DPPH2,2-Diphenyl-1-picrylhydrazyl
ABTS2,2′-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid)
CSIRCouncil of Scientific and Industrial Research
FCFolin–Ciocalteu
Na2CO3Sodium carbonate
GAEGallic acid equivalents
QEQuercitin equivalents
ANOVAAnalysis of variance
CRDCompletely randomized design
HATHydrogen atom transfer
ETElectron transfer
IC50Inhibitory concentration
RHSRoyal Horticultural Society

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Ijpb 14 00076 g001
Figure 1. Fully expanded detached rose leaves: (A) Louis Estes; (B) Thelma Barlow; (C) Le Rouge et le Noir; (D) Whippet; (E) Silver Shadow; (F) Dr. N.C. Sen; (G) Swamy; (H) Ma Normandie; (I) Majestic Burgundy; and (J) Grand Amore.
Figure 1. Fully expanded detached rose leaves: (A) Louis Estes; (B) Thelma Barlow; (C) Le Rouge et le Noir; (D) Whippet; (E) Silver Shadow; (F) Dr. N.C. Sen; (G) Swamy; (H) Ma Normandie; (I) Majestic Burgundy; and (J) Grand Amore.
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Figure 2. Variation in the total phenolic and flavonoid content in rose varieties.
Figure 2. Variation in the total phenolic and flavonoid content in rose varieties.
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Figure 3. Variation in L*, a*, and b* color values (adaxial surface) in different rose varieties.
Figure 3. Variation in L*, a*, and b* color values (adaxial surface) in different rose varieties.
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Figure 4. Variation in L*, a*, and b* color values (abaxial surface) in different rose varieties.
Figure 4. Variation in L*, a*, and b* color values (abaxial surface) in different rose varieties.
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Table 1. Color of leaves (RHS color chart) of different rose varieties used in study.
Table 1. Color of leaves (RHS color chart) of different rose varieties used in study.
VarietiesAdaxial Surface (Upper)Abaxial Surface (Lower)
Thelma BarlowGreyed purple group 187 AGreyed purple group 187 B
Le Rouge et le NoirGreyed purple group 187 AGreyed purple group 184 B
Majestic BurgundyGreyed purple group 187 ABrown group 200 C
Grand AmoreBrown group 200 AGreyed purple group 187 C
SwamyGreyed purple group 187 AGreyed purple group 187 C
Silver ShadowGreyed purple group 187 AGreyed purple group 187 C
Louis EstesGreyed purple group 183 AGreyed purple group 183 B
Dr. N.C. SenGreyed purple group 187 AGreyed purple group 184 A
WhippetGreyed purple group 187 AGreyed purple group 187 C
Ma NormandieBrown group 200 AGreyed purple group 184 A
Table 2. Variation in chlorophyll “a”, chlorophyll “b”, and total chlorophyll content in rose varieties.
Table 2. Variation in chlorophyll “a”, chlorophyll “b”, and total chlorophyll content in rose varieties.
Rose VarietiesChlorophyll “a”
mg/g		Chlorophyll “b”
mg/g		Total Chlorophyll
mg/g
Thelma Barlow0.51 ± 0.08 f0.17 ± 0.97 f0.74 ± 0.75 f
Le Rouge et le Noir0.79 ± 0.09 ab0.66 ± 0.84 b0.83 ± 0.25 e
Majestic Burgundy0.74 ± 0.32 c0.24 ± 0.08 d0.96 ± 0.84 d
Grand Amore0.80 ± 0.45 a0.74 ± 0.37 a1.41 ± 0.08 a
Swamy0.73 ± 0.08 c0.19 ± 0.09 e0.96 ± 0.98 d
Silver Shadow0.55 ± 0.98 e0.17 ± 0.86 f1.27 ± 1.32 b
Louis Estes0.76 ± 0.07 bc0.26 ± 1.23 c0.95 ± 0.97 d
Dr. N.C. Sen0.63 ± 0.08 d0.18 ± 0.08 f1.13 ± 0.42 c
Whippet0.63 ± 0.09 d0.18 ± 0.08 ef0.96 ± 0.06 d
Ma Normandie0.76 ± 0.78 bc0.28 ± 0.92 c0.84 ± 0.48 e
CD (0.05)0.0320.0170.055
Means followed by the same letter within a column were not significantly different at a 5% level of significance by t-test. The data show three replicates of each of the ten varieties.
Table 3. Variation in carotenoids and total anthocyanin content of rose varieties.
Table 3. Variation in carotenoids and total anthocyanin content of rose varieties.
Rose VarietiesCarotenoids
mg/g		Total Anthocyanin
mg/100 g
Thelma Barlow12.93 ± 1.36 h128.85 ± 0.40 a
Le Rouge et le Noir34.74 ± 0.94 b45.90 ± 0.08 i
Majestic Burgundy27.17 ± 0.87 d73.90 ± 0.08 f
Grand Amore36.29 ± 1.21 a35.40 ± 1.25 j
Swamy24.80 ± 1.66 e79.95 ± 1.38 e
Silver Shadow17.76 ± 0.93 g112.6 ± 0.98 b
Louis Estes28.12 ± 0.91 d59.25 ± 0.31 g
Dr. N.C. Sen18.28 ± 0.85 g94.20 ± 0.71 c
Whippet22.53 ± 0.89 f87.50 ± 0.53 d
Ma Normandie30.81 ± 0.08 c52.35 ± 0.58 h
CD (0.05)1.5141.459
Means followed by the same letter within a column were not significantly different at a 5% level of significance by t-test. The data show three replicates of each of the ten varieties.
Table 4. Variation in antioxidant activity (DPPH and ABTS) in rose varieties.
Table 4. Variation in antioxidant activity (DPPH and ABTS) in rose varieties.
Rose VarietiesDPPH IC50 (µg/mL)ABTS IC50 (µg/mL)
Thelma Barlow206.86 ± 0.49 a301.62 ± 2.31 a
Le Rouge et le Noir119.30 ± 0.95 i217.42 ± 2.02 h
Majestic Burgundy156.91 ± 0.79 f240.06 ± 2.33 f
Grand Amore111.30 ± 1.09 j177.58 ± 1.95 i
Swamy168.82 ± 0.19 e262.41 ± 2.60 e
Silver Shadow193.06 ± 0.44 b291.08 ± 2.68 b
Louis Estes149.00 ± 0.08 g223.14 ± 3.10 g
Dr. N.C. Sen181.19 ± 1.02 c282.73 ± 2.84 c
Whippet171.78 ± 2.04 d266.37 ± 2.63 d
Ma Normandie127.34 ± 0.08 h219.38 ± 0.69 h
CD (0.05)1.7544.584
Means followed by the same letter within a column were not significantly different at a 5% level of significance by t-test. The data show three replicates of each of the ten varieties.
Table 5. Correlation coefficient (r) between total anthocyanin content, total phenolic content, total flavonoid content, and antioxidant activity (DPPH, ABTS) in leaves of 10 rose varieties.
Table 5. Correlation coefficient (r) between total anthocyanin content, total phenolic content, total flavonoid content, and antioxidant activity (DPPH, ABTS) in leaves of 10 rose varieties.
ParametersTACTPCTFCDPPHABTS
TAC10.952 *0.969 *0.984 *0.968 *
TPC**10.975 *0.907 *0.871 *
TFC****10.943 *0.936 *
DPPH******10.969 *
ABTS********1
** = no correlation, * = positive correlation at a 5% level of significance by the t-test.
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Sharma, S.; Awasthi, L.; Kumari, P. Evaluating the Phytochemical Composition and Antioxidant Activity of Leaves of Different Rose Varieties. Int. J. Plant Biol. 2023, 14, 1051-1063. https://doi.org/10.3390/ijpb14040076

AMA Style

Sharma S, Awasthi L, Kumari P. Evaluating the Phytochemical Composition and Antioxidant Activity of Leaves of Different Rose Varieties. International Journal of Plant Biology. 2023; 14(4):1051-1063. https://doi.org/10.3390/ijpb14040076

Chicago/Turabian Style

Sharma, Shivani, Lipakshi Awasthi, and Poonam Kumari. 2023. "Evaluating the Phytochemical Composition and Antioxidant Activity of Leaves of Different Rose Varieties" International Journal of Plant Biology 14, no. 4: 1051-1063. https://doi.org/10.3390/ijpb14040076

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