Chemical Profiling and Antioxidant,Anti-Inflammatory,Cytotoxic,Analgesic,and Antidiarrheal Activities from the Seeds of Commonly Available Red Grape ( Vitis vinifera L.)

Abstract

Objectives:

The current study aimed to conduct a phytochemical screening of commonly known fruit red grape (Vitis vinifera L.) seed methanolic extract through gas chromatography and mass spectrometry (GC-MS) to identify the bioactive compounds responsible for its health benefits and evaluate the pharmacological potentialities of the extract and its fractions against oxidation, inflammation, pain, and diarrhea.

Methods:

The in vitro antioxidant, anti-inflammatory, and cytotoxic characteristics of methanolic extracts and various solvent fractions of V. vinifera were evaluated using the DPPH free radical scavenging assay, membrane stabilizing, and brine shrimp lethality bioassay. Furthermore, the study assessed the effects of crude extracts (200, 400, and 600 mg/kg of body weight) on pain relief and reduction of diarrhea in animals using methods such as tail immersion, the acetic acid-induced writhing technique, and a diarrheal mouse model induced with castor oil.

Results:

A total of 73 phytoconstituents were predominantly found in the seed extract based on the GC-MS analysis. Among the identified compounds, 9-octadecenamide (13.7%), and (9E,11E)-octadeca-9,11-dienoate (11.07%) are most abundant. Several notable constituents, such as gamma-sitosterol, stigmasterol, paromomycin, 4,6-cholestadienol, gamma-tocotrienol, 24-Propylidenecholest-5-en-3beta-ol, and alpha-tocopherol acetate, are also present. The methanolic extract of V. vinifera seed and its different solvent fractions showed promising antioxidant properties (IC₅₀ = 1.19-17.42 µg/mL) compared to the standard antioxidant butylated hydroxytoluene (IC₅₀ = 20.46 µg/mL). Aqueous soluble fraction exerted inhibition of nearly 50% heat-induced hemolysis compared to the standard acetylsalicylic acid (42%). Besides, all the tested doses (200, 400, and 600 mg/kg bw) of the crude extract showed significant (P < .05) analgesic and antidiarrheal effects.

Conclusion:

The current findings endorsed the health benefits of V. vinifera by revealing potent antioxidant, anti-inflammatory, analgesic, and antidiarrheal effects. Nevertheless, further in-depth analysis of the plant’s chemical constituents and pharmacological effects on health is warranted for novel drug discovery from V. vinifera.

Keywords

Red grape seed GC-MS analysis antioxidant anti-inflammatory cytotoxicity analgesic antidiarrheal

Introduction

Fruit is classified as a botanical product originating from the reproductive structures of plants, constituting a vital component of the human and animal diet. Fruits serve as a primary energy source while offering essential nutrients and an array of bioactive compounds crucial for maintaining optimal health. Furthermore, fruits exhibit health-promoting properties, mitigating susceptibility to particular ailments and age-related functional degenerations.¹ A significant correlation between a diet rich in antioxidants and diminished susceptibility to chronic illnesses was established in various research.² Cereals, legumes, oilseeds, as well as fruits and vegetables, serve as primary sources of dietary polyphenols and numerous bioactive constituents.^3,4 These components play pivotal roles in enhancing the functional and nutraceutical qualities of the diet.³ Moreover, it is noteworthy that not only the consumable portions of fruits but also the residual byproducts amount contain substantial well-known for of phenolic compounds and bioactive phytochemicals. These compounds are their notable antioxidant, anti-inflammatory, anti-mutagenic, and anti-carcinogenic properties.^3,5,6

Food loss and waste have emerged worldwide, manifesting at different points along the food supply chain. In 2019, hunger or insufficient access to nutritious food affected around 2 billion people, equivalent to about a quarter (25.9%) of the global population. Conversely, each year, about one-third (1.3 billion metric tons) of the food produced for human consumption goes to waste throughout the food supply chain.⁷ This wastage also entails substantial nutritional losses, with foods rich in nutrients like fruits and vegetables being the most commonly discarded. Consequently, reducing or reallocating food waste has the potential to enhance food accessibility and simultaneously elevate nutritional and dietary standards.

The most common fruits consumed globally encompass apples, grapes, and exotic fruits indigenous to their respective cultivation regions. Grapes notably constitute the largest fruit crop worldwide, boasting an annual production exceeding 75 million tons globally.⁸ Within this framework, grapes and their derivatives, including wine, grape juice, and preserves, have significant economic importance and exert a substantial influence on waste generation. Residues such as marc, peels, and grape seeds, persist due to inadequate management practices, thereby posing a contamination risk.⁸ Conversely, grape byproducts serve as a valuable reservoir of essential nutrients, encompassing vitamins, minerals, lipids, proteins, carbohydrates, and polyphenolic compounds.⁶

The red grape (Vitis vinifera L.; Family: Vitaceae), which grows in the Mediterranean and Central Asia, is a commonly consumed fruit rich in nutrients, which offers many health advantages.⁹ However, the winemaking sector is renowned for producing significant quantities of byproducts, and managing their disposal presents economic and environmental challenges. This issue of food waste has drawn considerable public health concern. Hence, there is value in conducting research to transform food waste into a usable resource and tackle worldwide nutritional deficiencies.^10,11

In several experimental investigations, red grape (Vitis vinifera L.) seed extract, which is a byproduct of manufacturing wine and is high in pro-anthocyanidins, has demonstrated promise. Research indicating its effectiveness against hypertension, inflammation, peptic ulcers, microbiological infections, cardiovascular disease, cancer, and diabetes has confirmed its pharmacological activity and beneficial health effects. Extracts of grape seeds are poised to be turned into a potential therapeutic due to their wide variety of applications.¹² In animal experiments, grape seed extract has demonstrated preventive properties, including suppression of DNA breakage, lipid peroxidation, and reduction of cardiac infarct size. It has been shown to inhibit lineages of human cancer cells in vitro.¹³

The current investigation utilizes GC-MS analysis to identify bioactive components from the methanolic extract of red grape (V. vinifera) seeds. This research additionally explored the pharmacological potentialities of the fruit seeds against oxidation, inflammation, pain, and diarrhea. These outcomes of the research may be helpful in spreading the nutraceuticals and medicinal value of the V. vinifera seeds, which is commonly available in Bangladeshi market.

Methods and Materials

Collection of V. vinifera seeds and drying

The Vitis vinifera (Family: Vitaceae), locally known as red grapes, were purchased from the local market of West Dhanmondi, Dhaka, Bangladesh, in January 2022. The batch was collected from Yantai, Shandong province, China. Following the acquisition of the fruit from the market, the process entailed the manual collection of fresh seeds, which were subsequently subjected to washing with fresh water. The seeds underwent a drying phase through natural shedding, facilitated by exposure to open air for an extended period. During this drying regimen, stringent measures were implemented to consistently maintain an environmental temperature below 30°C. This precautionary measure was adopted to ensure the preservation of heat-sensitive compounds and to preempt any possibility of degradation. Supplemental Figure S1 illustrates the dried seeds of V. vinifera.

Chemicals and reagents

Reagents of Analytical-level and substances were utilized in this investigation. All the chemicals such as Tween 80, tert-butyl-1-hydroxytoluene (BHT), Folin-Ciocalteau reagent, and Gallic acid were collected through Merck (Germany). Sigma-Aldrich (USA) supplied 2,2-diphenyl-1-picrylhydrazyl (DPPH). BEXIMCO Pharmaceuticals Ltd., Dhaka, Bangladesh, delivered the glibenclamide, normal saline solution, and loperamide as gift samples.

Extraction

A high-capacity grinding apparatus was employed to transform the desiccated seeds into a coarse powder. The extraction process followed the solid-liquid extraction method. It is noted that 500 g of powdered seeds were placed inside a sanitized amber bottle with a 3.0 L capacity. Subsequently, the powder was immersed in 2.0 L of methanol and allowed to soak for a period of 15 days at room temperature, with intermittent shaking and stirring. After that, a fresh cotton pad and filter paper were used to filter the solvent mixture. In order to produce a concentrated crude extract, the methanol was subsequently separated using an “EYELA Rotavapor” rotary evaporator at smaller pressures and about 40 to 50°C. Ultimately, a quantity of 67 g of gummy exudate was acquired from V. vinifera, signifying a yielding rate of 13.4%.

Fractionation

For the solvent-solvent differentiation of the extract, S. Morris Kupchan’s partitions (1970) technique, modified by VanWagenenet et al¹⁴ as used. It was accomplished by extracting 5 g of crude extract with water in methanol (1:9), petroleum ether (PESF), dichloromethane (DCMSF), ethyl acetate (EASF), and water (AQSF) to produce 4 distinct fractions. Each fraction also continued to evaporate until it was completely dried. The yields for the fractions produced by petroleum ether, dichloromethane, ethyl acetate, and water solubility were 25%, 24%, 18%, and 27%, respectively.

Gas Chromatography Mass Spectrometry (GC-MS) analysis

With an auto-sampler, Shimadzu, Japan, GC/MS-QP2010 ultra was used to analyze the phytoconstituents of the V. vinifera seeds’ methanolic extract. In a 5 MS/HP column (30 m, 0.25 mm, and 0.25 m), extremely pure helium was used as the mobile phase. The linear speed of the helium was 39 cm/s, and the helium circulation rate was 1.12 mL/min. The oven temperature was kept constant at an average rate of 10°C per minute, ranging from 110°C to 280°C. The needle temperature was adjusted to 250°C. The injection volume (50 µL) was entered in splitless mode (ratio of 10:1). The detector voltage was maintained at 0.94 kV. The ambient temperatures of the ion source, MS transfer line, and the ion source were all kept at 200 and 250°C, respectively. Full-scan mass spectra with a (m/z) range of 85 to 500 were captured at 10 000 u/s. Searching in the National Institute of Standards and Technology (NIST) collection yielded to identify peaks and chemical constituents.

Antioxidant properties

Total phenolic content (TPC)

The TPC of the seeds’ methanolic extract and fractions was determined using the Folin-Ciocalteu method.¹⁵ One milliliter of each fraction and extract (2 mg/mL) was combined with Na₂CO₃ (2.5 mL, 7.5% w/v) and a 10-fold diluted Folin-Ciocalteu reagent (2.5 mL). The solution was incubated in the dark at room temperature for 30 minutes. A UV-Vis spectrophotometer (760 nm) was used to test the sample’s absorbance. The calibration standard curve (Gallic acid) was created by placing the absorbance against various Gallic acid concentrations (100, 50, 25, 12.5, 6.25, 3.12, 1.56, 0.78, and 0.39 µg/mL) to calculate the TPC of the samples stated on the label as GAE mg (Gallic acid equivalent)/g of the study’s samples. The Gallic acid calibration curve formulations are as follows:

y = 0.0162 x + 0.0215 {, R}^{2} = 0.9985

DPPH scavenging assay

Antioxidant activity was qualitatively and quantitatively assessed using thin-layer chromatography (TLC) and the stable free radical 0.04% DPPH (1,1-diphenyl-2-picrylhydrazyl).¹⁶ To separate the polar and non-polar components of the extract, diluted stock solutions were applied to a dyed silica gel TLC plate, which was then placed in chambers containing solvents of varying polarities (polar, medium polar, and non-polar). The plate was subsequently sprayed with 0.02% DPPH in ethanol and incubated at room temperature. Following the DPPH treatment, color shifts (yellow on a purple background) were observed on the resolved bands for 10 minutes.¹⁶

The DPPH approach was employed to assess the antioxidant potential of MECE and various fractions.¹⁷ Initially, 3.0 mL of freshly made methanol and DPPH solution (20 µg/mL) was thoroughly combined with 2 mL of the samples’ various strengths (500, 250, 125, 62.5, 31.25, 15.62, 7.81, 3.9, 1.93, and 0.97 µg/mL) before being placed in a dark environment at room temperature for 30 minutes. The samples’ absorbance was then measured with a UV-Vis spectrophotometer calibrated to 517 nm. The inhibition percentage (I%) of the DPPH free radical was calculated using the following formula¹⁷:

I % = (A_{control} {- A}_{sample}) {/A}_{control} 100

A_control indicated the absorbance of the solution that had all reagents but lacked the test components. A_sample indicated the absorbance of the samples being examined or the standard (BHT) solution. Every test sample generated a graph showing the percentage of inhibition versus the concentrations of the tested substances, and the IC₅₀ (half concentration of the greatest inhibitory concentration) values were then calculated using a linear regression approach.

Anti-inflammatory effects

The effectiveness of V. vinifera seed extract and fractions to stabilize human erythrocyte membranes was investigated using hypotonic and heat-induced techniques.^18,19

Hypotonic solution-induced hemolysis

Red blood cells (RBCs) were given to a 70 kg adult with fair skin and no hidden disorders. The RBCs were put in a sterile container containing an anticoagulant, EDTA. A buffer solution with a pH of 7.4 was created using disodium phosphate and its conjugate acid, monosodium phosphate. 4.5045 g of NaCl was dissolved in sterile distilled water to make an isotonic solution (500 mL, 154 mM). 1.4625 g of NaCl was dissolved in sterile distilled water to make a hypotonic solution (500 mL, 50 mM). The blood was cleaned 3 times with sodium phosphate buffer and isotonic solution before centrifugation (3000 rpm, 10 minutes) to get the erythrocyte suspension. A stock RBC suspension (0.50 mL), a buffer of 10 mM sodium phosphate (4.5 mL), and a hypotonic solution (50 mM NaCl) were used as test samples, with different proportions of MECE (2.0 mg/mL) or standard ASA (0.10 mg/mL) as the reference standard. Before centrifugation (3000 rpm, 10 minutes), the resulting solutions were incubated at ambient temperature for 10 minutes. The supernatant’s absorbance (optical density (OD)) was then measured at 540 nm. The following calculation was used to compute the hemolysis inhibition percentage or membrane stabilization of the tested samples and standard ASA:

\begin{array}{l} % Hemolysis inhibition \\ (hypotonic solution induced) = 100 \times ({OD}_{1} {-OD}_{2}) {/OD}_{3} \end{array}

OD₁: hypotonic buffered saline solution optical density (Control)

OD₂: optical density of the examined sample in hypotonic solution

Heat-induced hemolysis

Two batches of centrifuge tubes containing 5 mL of isotonic buffer and 1.0 mg/mL of seed extract and various fractions were made. One tube with the same amount of material was used as a control. Each tube was filled with 30 µL of erythrocyte suspension and stirred slowly by inversion. A single set of tubes was submerged in a 54°C water bath for 20 minutes, whereas the additional set was maintained in a 0 to 5°C cold bath. Following incubation, the supernatant was turned to a centrifuge (1300 rpm, 3 minutes) and the optical density (OD) was measured at 540 nm. The percentage inhibition of hemolysis in the study was estimated using the following equation:

\begin{array}{l} % hemolysis \\ inhibition (heat-induced) = 100 \times (\begin{array}{l} 1 - {OD}_{2} - {OD}_{1} / \\ {OD}_{3} - {OD}_{1} \end{array}) \end{array}

Here, OD₁ = optical density (OD) of the test sample unheated, OD₂ = OD of test sample heated, and OD₃ = OD of control sample heated.

Cytotoxicity: brine shrimp lethality bioassay

The methanolic extract and its fractions of V. vinifera seeds were assessed for their cytotoxic effects through a brine shrimp lethality test described by Rashid et al,¹⁹ utilizing vincristine sulfate (VS) as a positive control. To create varying concentrations (ranging from 400.0 to 0.781 µg/mL) of the samples, a serial dilution of each 4 mg test sample was prepared in 99% DMSO (dimethyl sulfoxide). These solutions were then exposed to simulated seawater containing around 10 live brine shrimp nauplii. After a 24-hour period, the surviving nauplii were scrutinized using a magnifying glass. The toxicity level of the shrimp in response to each concentration of the sample was determined to estimate the LC₅₀ value. The LC₅₀ value, representing the concentration at which 50% of the shrimp were not viable, was calculated from a graph plotting the percentage of non-viable shrimp against the logarithmic concentration of the plant extract, utilizing the vincristine standard curve as reference.

Experimental design and sample size determination for in vivo studies

Swiss albino adult mice that were healthy, weighed between 25 and 30 g, and were between 4 and 5 weeks’ old were collected from the Animal Division of the International Center for Diarrheal Diseases and Research in Bangladesh (ICDDRB). The mice were kept in polypropylene cages under standard climatic settings, including a 25°C ambient temperature, a 55°% relative humidity level, and a 12-hour light/dark cycle before the animal study began. During this time, ICDDRB prepared food and water for them to consume. The study followed the rules established by the Federation of European Laboratory Animal Science Associations (FELASA) for the moral treatment of animals in investigations. The Animal Ethics Committee of State University of Bangladesh also carefully examined and confirmed the research’s ethical standards and procedures. In every test, mice were divided into 5 distinct groups; negative control, positive control, and 3 different (200, 400, and 600 mg/kg bw) sample groups I-III. Every group had 2 male and 2 female mice. Since we could not determine the standard deviation and effect size, we employed the “resource equation” method outlined by Mukta et al⁵ and Bulbul et al²⁰ to estimate the sample size. While working with a 1-way analysis of variance (ANOVA), we calculated the sample size for the degrees of freedom associated with the variability between subjects, also known as within-subject degrees of freedom, using the formula:

n = DF/ k + 1

In this formula, “n” represents the number of animals in each group, “DF” signifies the degrees of freedom, and “k” denotes the total number of groups. We considered the allowable range for degrees of freedom (DF) to establish the minimum and maximum number of animals per group. We obtained the respective minimum and maximum numbers of animals per group by using the minimum (10) and maximum (20) values for DF. In this case, we excluded the normal control group by setting ’k’ to 4, indicating 4 groups (comprising 3 test groups I-III with doses of 200, 400, and 600 mg/kg bw, and 1 positive control group with the standard drug). Therefore, the minimum and maximum numbers of animals per group should be 4 (10/k + 1 = 10/4 + 1) and 6 (20/k + 1 = 20/4 + 1), respectively. Following these ethical principles, we carried out this preliminary investigation with the minimal number of animals per group (n = 4). At the end of the study, according to the AVMA (American Veterinary Medical Association) guidelines for the Euthanasia of Animals, all mice were compassionately euthanized while under general anesthesia.²⁰

Central analgesic activity

The central analgesic effects of the MECE were evaluated using a heat approach and the tail immersion experiment.^21,22 The given morphine (15 mg/mL) was diluted with saline water to form the standard sample (subcutaneous, 2 mg/kg). The test medications were orally administered to the mice using a feeding syringe. The test included submerging a mouse’s tail in water heated to 55°C. At 0, 30, 60, and 90 minutes after giving the test samples to each mouse, the PRT or latency duration to move its tail in hot water was assessed.

Peripheral analgesic activity

The acetic acid-induced writhing method was employed to assess the peripheral analgesic effects of MECE.²² Each animal group received glacial acetic acid, a substance that induces pain. Tween 80 was used as a negative control, Diclofenac sodium (50 mg/kg) as a positive control, and MECE (200, 400, and 600 mg/kg) were given to the distinctive mouse groups. Acetic acid is in charge of writhing. The number of writhes was counted after delivering acetic acid intraperitoneally for 10 minutes. The following percentage of writhing inhibition was calculated:

Writhing inhibition % = (N_{Control} - N_{test}) {/N}_{Control} \times 100 %

where N = average amount of stomach writhing for every group.

Anti-diarrheal activity

A castor oil-induced diarrhea model in mice was implemented to evaluate the anti-diarrhea activity of MECE, as stated by earlier investigation.²³ Each of the 3 groups—the control, positive control, and test groups—contained 4 mice each. The control group received 1% Tween 80 in normal saline as a carrier solution and only orally at a specific dose (10 mL/kg). While different doses (200, 400, and 600 mg/kg) of MECE were provided to the 3 test groups with Tween 80 (1% in normal saline) as a carrier, loperamide (50 mg/kg) was administered orally to the positive control group. Each mouse was administered 1 mL of pure castor oil to induce diarrhea after 1 hour. Each mouse was housed in an individual cage placed on the floor, lined with blotting paper. The quantity of diarrheal stools for each mouse was recorded at the conclusion of every hour for up to 4 hours following the administration of castor oil. The blotting paper was replaced at the beginning of each hour. Observations from the test groups were compared with those from the negative and positive control groups to evaluate the potential of MECE as an antidiarrheal agent. The percentage reduction in diarrhea was calculated using the following formula to assess the antidiarrheal activity.²³

% inhibition of defecation = (D_{control} - D_{test}) {/D}_{control} \times 100 %

where D = Average diarrheal episode/number in every group.

Statistical analysis

MS Excel (version 10.0) was used for processing the graphs and data related to in vitro investigation. Because the in vitro data was derived from a single experiment, there was no statistical analysis conducted for comparing between groups; instead, a numerical comparison was carried out. Instead, statistical analysis was done on the data collected from the in vivo tests. We compared the treatment groups to the control (vehicle) group. The average values with their associated standard errors of the mean was expressed as mean ± SEM to summarize the in vivo results. To analyze the in vivo data, we used Statistical Package for Social Sciences (SPSS) Software (version 26.0), conducting a 1-way analysis of variance (ANOVA) on all variables, followed by a student’s t-test. Any P-values below .05 were considered statistically significant.

Results

Identification of phytoconstituents by GC-MS analysis

A total of 73 phytoconstituents were predominantly found in the MECE based on their GC-MS analysis. The spectrum obtained from GC-MS analysis is available as Figure 1. Among the isolated compounds, 9-Octadecenamide (13.7%), (9E,11E)-octadeca-9,11-dienoate (11.07%), 1,3-dihydroxypropan-2-yl(9Z,12Z)-octadeca-9,12-dienoate (9.06%), and Tris(hydroxymethyl)nitromethane (7.93%) were most abundant (Table 1), while squalene (0.08%), 13-Docosenoicacid, methyl ester (Z)- (0.1%), and Vitamin E (0.1%) were the least amount compounds (Table 1). Some bioactive plant steroids such as gamma-sitosterol, stigmasterol, Ergost-5-en-3-ol, (3beta, 24R)-, and 24-Propylidenecholest 5-en-3beta-ol were also detected from the MECE. In addition, some notable compounds are paromomycin, 4,6-cholestadienol, gamma-tocotrienol and alpha-tocopherol acetate. According to GC-MS analysis, all identified compounds’ names, retention time (RT), % area, molecular formula, molecular weight, PubChem CIDs, and chemical structures were tabulated in Table 1. In addition, the reported bioactivities of these identified compounds were also tabulated with their corresponding references (Table 1).

Table 1.

List of Detected Constituents by Gas Chromatography Mass Spectrometry (GC-MS) from Methanolic Extract of V. vinifera.

Sl. no.	Compound	RT	Area %	Mol. weight (g/mol)	Molecular formula	PubChem CID	Biological activity	Ref
1	Pentanedioic acid	5.027	0.39	132.11	C₅H₈O₄	743	Anti-neoplastic activity	Goswami et al²⁴
2	Decyl tetradecyl ester carbonic acid	5.105	0.32	272.423	C₂₅H₅₀O₃	91693142	Antioxidant, Antibacterial activity	Gopal Pandit et al²⁵
3	2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one	5.365	0.82	144.12	C₆H₈O₄	538757	Anti-oxidative	Cerny²⁶
4	Tranylcypromine	5.41	0.78	133.19	C₉H₁₁N	5530	MAO inhibitor	Tranylcypromine²⁷
5	1-(furan-2-yl)-2-methylprop-2-en-1-one	5.532	4.56	136.15	C₈H₈O₂	3007408	Anti-fungal activity	Ogata et al²⁸
7	Limonene	5.817	1.46	136.23	C₁₀H₁₆	22311	Anti-bacterial activity	Han et al²⁹
8	Maple lactone	5.91	0.55	112.12	C₆H₈O₂	6660	Anti-oxidant activity	Kim et al³⁰
10	2-amino octanoic acid	6.09	1.36	159.22	C₈H₁₇NO₂	69522	Antibiotic activity	Almahboub et al³¹
11	Furaneol	6.251	1.46	128.13	C₆H₈O₃	19309	Anti-microbial & Anti-fungal activity	Sung et al³²
12	1,3-Propanediol, 2-methyl-2-propyl	6.435	0.26	370.38	C₁₁H₂₂N₄O₈S	117593	Sedative, anticonvulsant and muscle relaxant effect	Yale et al³³
13	Thymine	6.489	1.19	126.11	C₅H₆N₂O₂	1135	Anti-cancer & Anti-microbial activity	Kumar et al³⁴
14	2-methoxy phenol	6.617	0.6	124.13	C₇H₈O₂	460	Antibacterial and antioxidant activity	Rubab et al³⁵
15	Nonane,5-(1-methylpropyl)	6.683	0.21	184.36	C₁₃H₂₈	43943	Antioxidant and Antimicrobial activity	Ashraf et al³⁶
16	Diazene, bis(1,1-dimethylethyl)-	6.725	0.61	142.24	C₈H₁₈N₂	70227	Anti-phytopathogenic	Singh et al³⁷
19	1,2,4-Benzenetriol	6.94	0.22	126.11	C₆H₆O₃	10787	Anti-oxidant	Zhang et al³⁸
21	2-Propanamine, N-methyl-N-nitroso	7.141	0.48	102.14	C₄H₁₀N₂O	92271	Anti-bacterial	Kadiri et al³⁹
22	Limonene oxide	7.197	0.3	152.23	C₁₀H₁₆O	8029780	Bactericidal, anticancer,	Junior and Pastore⁴⁰
23	2,3-Dihydro-3,5-dihydroxy-6-methyl-4h-pyran-4-one	7.282	2.56	144.12	C₆H₈O₄	119838	Anti-oxidant and mutagenic	⁴¹
24	4 chloroanisole	7.459	0.49	142.583	C₇H₇ClO	11137567	Antibacterial activity	Boubakri et al⁴²
25	5-methyl-4H-1,2,4-triazol-3-amine	7.545	0.2	98.11	C₃H₆N₄	234610	Antiviral & Anti-infective activity	⁴³
28	Catechol	7.81	2.88	110.11	C₆H₆O₂	289	Anti-microbial, Anti-fungal activity	Kocaçalışkan et al⁴⁴
29	2-methyle-5(1-methylethyl)-cyclohexanone	7.957	0.63	154.25	C₁₀H₁₈O	10362	Weak fungicidal activity	Xia et al⁴⁵
30	(1R,5R)−2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol	8.09	1.86	152.23	C₁₀H₁₆O	11084068	Anti-bacterial & Anti-fungal activity	Aberchane et al⁴⁶
31	5-(hydroxymethyl)furan-2-carbaldehyde	8.152	0.86	126.11	C₆H₆O₃	237332	Anti-oxidant & Anti-proliferative	Qiu et al⁴⁷
32	Carveol	8.23	0.36	152.23	C₁₀H₁₆O	7438	Anti-oxidant, Anti-cancer & vasorelaxation	Lacerda-Neto et al⁴⁸
35	cis-Limonene oxide	8.638	1.05	152.23	C₁₀H₁₆O	6452061	Anti-microbial & insecticidal activity	He et al⁴⁹, Aggarwal et al⁵⁰
36	4-Methyl catechol	8.807	0.88		C₇H₈O₂	9958	Antioxidant	Ito et al⁵¹
37	o-Tolunitrile	8.91	0.13	117.15	C₈H₇N	10721	NA
38	Cyclopropane	8.94	0.16	42.08	C₃H₆	6351	NA
39	Decyl 2-chloroacetate	8.975	0.18	234.76	C₁₂H₂₃ClO₂	229381
40	2-Methoxy 4-vinylphenol	9.046	0.29	150.17	C₉H₁₀O₂	332	Antimicrobial, antioxidant, anti-inflammatory, analgesic, anti-germination	Ibibia et al⁵²
41	3-(Methylthio)propyl acetate	9.165	0.13	148.23	C₆H₁₂O₂S	85519	Flavor activity	Aoki and Uchida⁵³
43	1-Methoxy-6,6-dimethylcyclohex-1-ene	9.24	0.18	140.22	C₉H₁₆O	580527	NA
44	9,12,15-Octadecatrienoic acid, methyl ester, (9Z,12Z,15Z)-	9.329	0.15	292.5	C₁₉H₃₂O₂	9316	Antioxidant, anti-carcinogenic, anti-inflammatory, and anti-obese	Yuan et al⁵⁴
45	1,2,3-Benzenetriol	9.646	7.39	126.11	C₆H₆O₃ or C₆H₃(OH)₃	1057	antibacterial and antioxidant	Cynthia et al⁵⁵
46	Paromomycin	10.2	0.22	615.6	C₂₃H₄₅N₅O₁₄	441375	Antibiotic activity	Davidson et al⁵⁶
48	Tris(hydroxymethyl)nitromethane	10.575	7.93	151.118	C₄H₉NO₅	31337	Preservative, Disinfectant & Bactericidal activity	Izzat and Bennett⁵⁷
49	Anhydro-d-mannosan	10.839	0.65	162.14	C₆H₁₀O₅	11947765	Muscle-relaxing and pain relieving	Yellu and Bhukya⁵⁸
50	2′-Hexyl-1,1”-bicyclopropane-2-octanoic acid methyl ester	10.93	0.34	322.5	C₂₁H₃₈O₂	552098	Anti-microbial activity	Srivastava et al⁵⁹
52	4-Propylresorcinol	11.842	1.73	152.19	C₉H₁₂O₂	87874	Anti-tyrosine activity	Matsubara et al⁶⁰
53	2-(hydroxymethyl)-6-octylsulfanyloxane-3,4,5-triol	12.04	0.4	308.44	C₁₄H₂₈O₅S	363418	NA
54	Ethylalpha-d-glucopyranoside	12.105	0.69	208.21	C₈H₁₆O₆	91694274	Preservative	Rajalakshmi and Mohan⁶¹
55	3-Deoxyhexonic acid	12.19	0.25	180.16	C₆H₁₂O₆	10350	NA
56	Dihydroconiferyl alcohol	12.256	0.39	182.22	C₁₀H₁₄O₃	16822	Stimulating cell growth	Lee et al⁶²
57	2-Hydroxy-5-methylisophthalaldehyde	14.311	0.28	164.16	C₉H₈O₃	81744	NA
58	(9E,11E)-octadeca-9,11-dienoate	18.337	11.07	279.4	C₁₈H₃₁O₂		Metabolic regulator, hypocholesterolemia, anti-atherogenic, anti-carcinogenic, antioxidant,	Fagali and Catalá⁶³
59	(E)-methyl octadec-11-enoate	18.436	1.95	296.5	C₁₉H₃₆O₂	74738	Anti-diarrheal activity
60	13-Docosenoicacid, methyl ester (Z)-	24.971	0.1	352.6	C₂₃H₄₄O₂	5363109	Anti-cancer activity	Paudel and Pant⁶⁴
61	3,3-dihydroxypropyl palmitate	25.108	2.24	689.1	C₄₀H₈₀O₈	24698	Anti-cancer	Zhu et al⁶⁵
62	1,3-dihydroxypropan-2-yl (9Z,12Z)-octadeca-9,12-dienoate	27.855	9.06	354.5	C₂₁H₃₈O₄	5365676	Analgesic, anti-inflammatory andAnti-ulcerogenic	Mohammed et al⁶⁶
63	2,3-Dihydroxypropyl 12-hydroxyoctadecanoate	28.288	0.55	374.6	C₂₁H₄₂O₅	95408	NA
64	9-Octadecenamide	29.195	13.7	281.5	C₁₈H₃₅NO	1930	Antioxidative and hypolipidemic properties	Cheng et al⁶⁷
65	Squalene	29.654	0.08	410.7	C₃₀H₅₀	638072	anticancer, antioxidant, drug carrier, detoxifier, skin hydrating, and emollient	Kim and Karadeniz⁶⁸
66	4,6-cholestadienol	33.363	0.13	384.6	C₂₇H₄₄O	14795191	Broad spectrum antimicrobial activity	Zhu et al⁶⁹
67	Vitamin E	34.093	0.1	430.7	C₂₉H₅₀O₂	14985	Antioxidant activity	Stanner and Weichselbaum⁷⁰
68	Gamma-Tocotrienol	34.698	0.13	410.63	C₂₈H₄₂O₂	5282349	Antitumor activity	Suman et al⁷¹
69	Ergost-5-en-3-ol, (3beta,24R)-	35.798	0.31	400.7	C₂₈H₄₈O	6428659	Antioxidant, Anti-cancerous properties	Kang et al⁷²
70	Alpha-tocopherol acetate	36	0.18	472.7	C₃₁H₅₂O₃	86472	antioxidant	Tucker and Townsend⁷³
71	Stigmasterol	36.234	0.68	412.7	C₂₉H₄₈O	5280794	Antimicrobial, anticancer,diuretic, anti-inflammatory,antioxidant	Khalid et al⁷⁴
72	Gamma sitosterol	37.372	3.54	414.7	C₂₉H₅₀O	457801	Anti-hyperglycemic	Sirikhansaeng et al⁷⁵
73	24-Propylidenecholest-5-en-3beta-ol	37.647	0.29	426.7	C₃₀H₅₀O	193212	Anti-bacterial	Kavita et al⁷⁶

Figure 1.

GC-MS spectrum of the crude extract obtained from V. vinifera L.

Assessment of antioxidant properties

Total phenolic content (TPC)

The findings revealed that the MECE expressed the most TPC (223.31 mg of GAE/g of dried extract), then the EASF and DCMSF fractions at 190.25 mg of GAE/g and 135.25 mg of GAE/g, respectively. In contrast, the PESF fraction indicated the least TPC, with 11.00 mg of GAE/g (Figure 2). These results suggest that the phenolic profiles of the various fractions of the seed extract are diverse, which may have consequences for their prospective bioactivity and use.

Figure 2.

Total phenolic content of the various fractions and methanolic extract of V. vinifera seeds. (Here, MECE = methanolic crude extract of V. vinifera, PESF = petroleum ether soluble fraction, DCMSF = dichloromethane soluble fraction, EASF = ethyl acetate soluble fraction, and AQ = aqueous or water-soluble fraction).

DPPH scavenging activity

In this study, the antioxidant potency of methanolic extract of V. vinifera seed and its different solvent fractions showed promising antioxidant properties (IC₅₀ = 1.19-17.42 µg/mL) compared to that of the standard antioxidant butylated hydroxytoluene (BHT; IC₅₀ = 20.46 µg/mL; Figure 3). The EASF has the highest free radical scavenging potency (IC₅₀ = 1.19 µg/mL), followed by MECE (IC₅₀ = 2.94 µg/mL), AQSF (IC₅₀ = 6.55 µg/mL), PESF (IC₅₀ = 16.81 µg/mL), and DCMSF (IC₅₀ = 17.42 µg/mL).

Figure 3.

DPPH free radical scavenging activity (IC₅₀, μg/mL) of various solvent fractions and methanolic extract of V. vinifera seeds. (Here, MECE = methanolic crude extract of V. vinifera, PESF = petroleum ether soluble fraction, DCMSF = dichloromethane soluble fraction, EASF = ethyl acetate soluble fraction, and AQ = aqueous or water-soluble fraction).

Anti-inflammatory effects

Hypnotic medium-induced hemolysis has been diminished as below: AQSF (22.62%), DCMSF (13.62%), PESF (10.39%), MESF (6.53%), and EASF (7.10%). The standard acetylsalicylic acid (ASA) inhibition rate was significant (61.90%; Figure 4). The most efficacious inhibitor of heat-induced hemolysis was AQSF (44.9%), followed by DCMSF (31.15%), PESF (21.65%), MESF (13.43%), and EASF (13.50%). The inhibition of the standard ASA was 42.00% (Figure 4).

Figure 4.

Membrane stabilizing impact of MECE and various fractions on hypotonic medium hemolysis and heat-induced hemolysis. (Here, MECE = methanolic crude extract of V. vinifera, PESF = petroleum ether soluble fraction, DCMSF = dichloromethane soluble fraction, EASF = ethyl acetate soluble fraction, and AQ = aqueous or water-soluble fraction).

Cytotoxicity

Each sample that underwent testing displayed noteworthy lethality against brine shrimp larvae, as evidenced by their LC₅₀ values (15.95-83.24 µg/mL), which were comparable to the established benchmark set by vincristine sulfate with an LC₅₀ of 0.45 µg/mL (Figure 5).

Figure 5.

Cytotoxicity of methanolic extract and its fractions of V. vinifera seeds in terms of LC₅₀ (µg/mL). (Here, MECE = methanolic crude extract of V. vinifera, PESF = petroleum ether soluble fraction, DCMSF = dichloromethane soluble fraction, EASF = ethyl acetate soluble fraction, and AQ = aqueous or water-soluble fraction).

Central analgesic activity

As stated in Figure 6, all the doses of the tested sample expressed significant (P < .05) dose- and time-dependent analgesic properties compared to standard morphine. The groups of ingesting MECE at various doses demonstrated a rise in pain reaction latency compared to the control, whereas the negative control possessed no analgesic impact.

Figure 6.

Central analgesic properties of crude methanolic extract of V. vinifera seeds. Values are expressed as Mean ± SEM (n = 4). ***P < .001, *P < .05 compared to negative control group. Positive control (morphine at 2 mg/kg b.w.), Groups I, II, and III = 200, 400, and 600 mg/kg bw = Methanol extract V. vinifera seed, respectively.

Peripheral analgesic activity

The study found that MECE showed noticeable and dose-dependent activity in peripheral analgesic assessment compared to the negative control group. The outcomes demonstrated that the seeds extracted from groups I to II exhibited a time- and dose-dependent reduction in writhing activity in mice (Figure 7). The seeds extract at group II and group III exerted 36.78% and 43.68% writhing inhibition in mice, around half of the standard diclofenac sodium (77.01% inhibition; Supplemental Figure S2). All the sequels were statistically significant with P < .05, indicating that the MECE might have a peripheral analgesic effect in mice in response to acetic acid-induced stomach writhing.

Figure 7.

Peripheral analgesic properties of crude methanolic extract of V. vinifera seeds. Values are expressed as Mean ± SEM (n = 4). ***P < .001, *P < .05 compared to negative control group. Positive control (Morphine at 2 mg/kg b.w.), Groups I, II, and III = 200, 400, and 600 mg/kg bw = Methanol extract V. vinifera seeds, respectively.

Anti-diarrheal activity

The results of the MECE indicated a statistically significant reduction in the total amount of diarrheal feces at all doses of MECE (Figure 8). Notably, the 400 mg/kg dose of MECE demonstrated the topmost degree of activity (80.43%) compared to the positive control Loperamide (78.26%) for the reduction of diarrhea (Supplemental Figure S3).

Figure 8.

Effects crude methanolic extract on the number of diarrheal feces (mean ± SEM) of mice after 4h of administration of castor oil. “***” means P < .001 compared to negative control group. Positive control (loperamide at 50 mg/kg b.w.), Groups I, II, and III = 200, 400, and 600 mg/kg bw = Methanol extract V. vinifera seeds, respectively.

Discussion

Herbal remedies, including parts like fruit, seeds, bark, fruit peel, and leaves, are frequently viewed as valuable sources of bioactive phytochemicals for addressing various health issues such as oxidative stress, diabetes, pain, fever, cancer, hypertension, and many more human illness.⁷⁷ The present research examined the chemical components within the seed extract of V. vinifera. This analysis revealed a range of potentially valuable bioactive elements. These elements could play a crucial role in driving the diverse bioactive effects observed in the red grape seed extract and its different solvent-based fractions. The GC-MS analysis of the MECE expressed the presence of several phytoconstituents, such as organic acids (pentanedioic acid, 2-amino octanoic acid), esters (Decyl tetradecyl ester carbonic acid), aliphatic hydrocarbons (Limonene, Nonane,5-(1-methylpropyl), Cyclopropane, Squalene), ketones (2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one, 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one, 1-(furan-2-yl)-2-methylprop-2-en-1-one, Maple lactone), alcohols (1,3-Propanediol,2-methyl-2-propyl, carveol), amine (Tranylcypromine), phenols (2-methoxy phenol, 1,2,4-Benzenetriol, Catechol, 4-Methyl catechol), steroids (gamma sitesterol, 24-Propylidenecholest-5-en-3beta-ol, stigmasterol, ergost-5-en-3-ol, (3beta, 24R)-), and tocopherols (alpha-tocopherol, gamma-tocotrienol; Table 1). These 73 compounds from the MECE may contribute to the medicinal activities of the fruit. From the previous literature study, most of the detected compounds in this study have anti-inflammatory, antioxidant, antibacterial, and anticancer effects (Table 1). The availability of cyclic unsaturated compound especially which has an aromatic ring such as catechol and 1,2,4-Benzenetriol may play a role for antioxidant properties of the MECE.^38,44 Another molecule squalene was also identified from this plant which have potential anticancer and antioxidant activities that are matched with the grape fruits bioproperties.⁶⁸

Free radicals and oxidants are harmful to the body because they are produced by both natural cellular processes and external causes such as emissions, smoking, medicines and radiation. The buildup of free radicals exceeds the body’s ability to remove them, resulting in oxidative stress. This mechanism has a crucial involvement in the onset of degenerative and chronic illnesses such as cardiovascular disease, autoimmune disorders, carcinoma, rheumatoid arthritis, cataracts, aging, and neurological diseases. The human body uses a variety of strategies to resist oxidative stress, including the creation of antioxidants. These antioxidants can be produced naturally by the body or obtained from food sources or supplementation.⁷⁷ In this study, the MECE showed the highest total phenolic content and EASF showed the highest scavenging capacity against DPPH freer radical. Zeghad et al⁷⁸ showed that whole fruits with seeds had tremendous antioxidant activity. Another study revealed that V. vinifera is the most abundant source of various flavonoids, polyphenols, and caffeic acid derivatives, while V. vinifera showed positive outcomes in the DPPH and ferric reducing antioxidant power (FRAP) assays.⁷⁹

Due to the harmful nature of a crude medicinal substance which is a significant worry, a low-cost and dependable method, the brine shrimp lethality bioassay using Artemia salina, was performed to initially screen for cellular toxicity in plant extracts. A study conducted by Lagartoparra et al⁸⁰ discovered a strong connection (with a correlation coefficient of .85 and a significance level of P < .05) between the 50% lethal concentration (LC₅₀) derived from the brine shrimp test and the 50% lethal dose (LD₅₀) from animal trials. This indicates that the brine shrimp test could serve as an alternative model. Meyer et al⁸¹ suggested that a bioactive plant compound would generally have an LC₅₀ value of <1000 μg/mL. The present research revealed that the LC₅₀ values obtained from the brine shrimp bioassay were all below 1000 μg/mL. As demonstrated by previous research (LC₅₀ >10 μg/mL), none of the plant’s crude extracts or fractions should be deemed extremely toxic or deadly.⁸²

The current research has also exerted a promising anti-inflammatory effect in heat-induced membrane stabilizing assays. Previous research has shown that V. vinifera seed extract has considerable anti-inflammatory effects by significantly reducing gene expression as well as protein secretion of inflammatory factors like tumor necrosis factor (TNF-α), interleukin-6 (IL-6), the inducible isoform of nitric oxide synthase (iNOS), and nitric oxide (NO).⁸³ Through histological examination, another study using mice found that seed extracts successfully diminished the levels of inflammatory molecules such as TNF-α, NF-K, IKK-a, IL-6, and IL-1.⁸⁴ However, the findings of this current study imply that the AQSF with heat-induced hemolysis showed a similar significance anti-inflammatory effect compared to the standard drug (44.99% versus 42.00%, respectively). In the GC-MS analysis, some anti-inflammatory compounds such as 2-Methoxy 4-vinylphenol, 9,12,15-Octadecatrienoic acid and 1,3-dihydroxypropan-2-yl (9Z,12Z)-octadeca-9,12-dienoate, and stigmasterol were detected, which may responsible for the anti-inflammatory properties of the MECE. Furthermore, large antioxidant molecules detection may also be responsible for the anti-inflammatory action of the fruit. The inflammation is mainly brought on by oxidative stress.⁸⁵ Histamine, serotonin, proinflammatory cytokines (including interleukin-1B and tumor necrosis factor-α), and inflammatory cells like leukotrienes and macrophages are the mediators involved in the complicated inflammation process.⁸⁵ Thromboxane A2, prostaglandins, and leukotrienes are examples of arachidonic acid metabolic products that also play a role in this process. Finally, these inflammations cause pain and diarrhea.⁸⁶ As a result, the presence of antioxidant molecules in V. vinifera is advantageous since they aid in controlling inflammation by neutralizing damaging ROS. These may help prevent or treat inflammatory-related disorders and positively impact general health.

Natural remedies for pain relief are being sought as replacements to manmade medications since they are associated with side effects.⁸⁷ In this study, we found that the seed extract of V. vinifera significantly decreased both central and peripheral pain experience in mice. Several analgesic compounds such as 2-methoxy 4-vinylphenol, Anhydro-d-mannosan, 1,3-dihydroxypropan-2-yl (9Z,12Z)-octadeca-9,12-dienoate were also recorded from the presence study. Moreover, the predominant antioxidant molecules also conclude the potential analgesic properties of MECE. Nadia et al showed that V. vinifera has dose-dependent central analgesic activity.⁸⁸ Aouey et al indicated in their study that some compounds mainly caffeoyltartaric acid and flavonoids derivatives were detected from the V. vinifera extract that may interfere the prostaglandins pathways.⁸⁹ Antioxidant molecules effectively inhibit the production of the prostaglandins and COX-2 production, which are responsible for the pain reaction.⁹⁰ In addition, endogenous fatty acid amides have been shown in previous research to exhibit significant bioactivity both centrally and peripherally.⁸⁷

The current study also found significant antidiarrheal effects of the MECE. Castor oil induces diarrhea primarily through 3 mechanisms: producing nitric oxide, increasing gastrointestinal membrane calcium permeability, and triggering prostaglandin production, resulting in increased fluid and electrolytes in the intestine, and stimulating peristalsis.⁹¹ The key ingredient in castor oil, ricinoleic acid, is alleged to upset the gut wall by generating prostaglandins and inducing peristaltic movement that might lead to diarrhea.^92-94 Antioxidants molecules have significant prostaglandin inhibition activity.⁹⁵ In this current study, although, no significant anti-diarrhea compounds were identified, several antioxidant compounds were detected from this GC-MS analysis, which may be responsible for the inhibition of castor oil-induced gut inflammation diarrhea effects.

Limitations and future research

The study presents certain areas for improvement, notably the need for comprehensive chromatographic isolation, purification, and spectroscopic characterization of the phytoconstituents from the investigated sample. Future research can be done to isolate pure compounds from the fruit’s seed extract and evaluate their pharmacological potential via in vitro, in vivo, and in silico analyses against diverse therapeutic targets.

Conclusion

The current study identified 73 phytoconstituents, including 9-octadecenamide, gamma-sitosterol, stigmasterol, paromomycin, 4,6-cholestadienol, gamma-tocotrienol, 24-Propylidenecholest-5-en-3beta-ol, and alpha-tocopherol acetate, from the seed extract of V. vinifera. The study also evaluated its pharmacological properties, focusing on antioxidant, anti-inflammatory, cytotoxicity, analgesic, and antidiarrheal activities. The methanolic seed extract and its various solvent fractions exerted promising antioxidant and anti-inflammatory properties, and the 3 tested doses (200, 400, and 600 mg/kg bw) of the crude extract showed significant in vivo effects against pain and diarrhea. However, further investigation is required to completely comprehend the precise processes and to discover the bioactive substances that regulate the actions of V. vinifera via in vitro, in vivo, and in silico approaches. Nevertheless, the antioxidant ability of this plant species provides hope for reducing inflammation and enhancing health.

Supplemental Material

sj-docx-1-nmi-10.1177_11786388241275100 – Supplemental material for Chemical Profiling and Antioxidant, Anti-Inflammatory, Cytotoxic, Analgesic, and Antidiarrheal Activities from the Seeds of Commonly Available Red Grape (Vitis vinifera L.)

Supplemental material, sj-docx-1-nmi-10.1177_11786388241275100 for Chemical Profiling and Antioxidant, Anti-Inflammatory, Cytotoxic, Analgesic, and Antidiarrheal Activities from the Seeds of Commonly Available Red Grape (Vitis vinifera L.) by Md. Jamal Hossain, Khadija Rahman Lema, Md Abdus Samadd, Rumi Aktar, Mohammad A. Rashid and Muhammad Abdullah Al-Mansur in Nutrition and Metabolic Insights

Footnotes

Acknowledgements

We would like to express our sincere appreciation to the Department of Pharmacy, State University of Bangladesh, for their kind assistance in assisting the research by providing laboratory facilities.

Author Contributions

JH: Conceptualization, Writing – original draft, Writing – review & editing, Visualization, Validation, Supervision, Software, Resources, Project administration, Methodology, Investigation, Formal analysis, Data curation. KRL: Visualization, Validation, Software, Resources, Methodology, Formal analysis, Data curation. AS: Writing – original draft, Visualization, Validation, Software, Resources, Methodology, Investigation, Formal analysis, Data curation. RA: Visualization, Validation, Software, Resources, Methodology, Formal analysis, Data curation. MAR: Writing – review & editing, Visualization, Validation, Project administration. MAA: Writing – review & editing, Visualization, Validation, Software, Resources, Data curation.

Declaration Of Conflicting Interests:

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding:

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Consent for Publication

Not applicable.

Data Availability Statement

The article contains all necessary information to support the conclusions. Contacting the corresponding author with a fair request will get you further raw data.

Ethical Approval

The work adhered to the standards for the ethical treatment of animals in research set forth by the Federation of European Laboratory Animal Science Associations (FELASA). The Animal Ethics Committee of the State University of Bangladesh thoroughly reviewed and approved the research’s ethical standards and practices.

ORCID iD

Md Jamal Hossain

Supplemental Material

Supplemental material for this article is available online.

References

Kumoro

Alhanif

Wardhani

DH.

A critical review on tropical fruits seeds as prospective sources of nutritional and bioactive compounds for functional foods development: a case of Indonesian exotic fruits. Int J Food Sci. 2020;2020:4051475.

Kaparapu

Pragada

Geddada

MN.

Fruits and vegetables and its nutritional benefits. In: Egbuna

Dable-Tupas

, eds. Functional Foods and Nutraceuticals: Bioactive Components, Formulations and Innovations. Springer, 2020;241-260.

Allaqaband

Dar

Patel

, et al Utilization of fruit seed-based bioactive compounds for formulating the nutraceuticals and functional food: A review. Front Nutr. 2022;9:902554.

Mitra

Lami

Uddin

, et al Prospective multifunctional roles and pharmacological potential of dietary flavonoid narirutin. Biomed Pharmacother. 2022;150:112932.

Mukta

Hossain

Akter

, et al Cardioprotection of water spinach (Ipomoea aquatica), wood apple (Limonia acidissima) and linseed (Linum usitatissimum L.) on doxorubicin-induced cardiotoxicity and oxidative stress in rat model. Nutr Metab Insights. 2023;16:11786388231212116.

Sarwar

Hossain

Irfan

, et al Renoprotection of selected antioxidant-rich foods (water spinach and red grape) and probiotics in gentamicin-induced nephrotoxicity and oxidative stress in rats. Life. 2022;12:60.

Brennan

Browne

Food waste and nutrition quality in the context of public health: A scoping review. Int J Environ Res Public Health. 2021;18:5379.

Fierascu

Sieniawska

Ortan

Fierascu

Xiao

Fruits By-Products – A source of valuable active principles. A short review. Front Bioeng Biotechnol. 2020;8:319.

Hussain

Naseer

Qadri

Fatima

Bhat

TA.

Grapes (Vitis vinifera)—Morphology, taxonomy, composition and Health Benefits. Fruits Grown in Highland Regions of the Himalayas: Nutritional and Health Benefits. Hussain

et al., eds. Springer International Publishing; 2021;103-115.

10.

Radulescu

Claudia Buruleanu

Lucian Olteanu

, et al Grape by-Products: potential sources of phenolic Compounds for novel functional foods. In: Bronze

, ed. Food Science and Nutrition. IntechOpen; 2023.

11.

Rojas

Castro-López

Sánchez-Alejo

Niño-Medina

Martínez-ávila

GCG

. Phenolic compound recovery from grape fruit and By- products: an overview of extraction methods. In: Grape and Wine Biotechnology. InTechOpen; 2016.

12.

Gupta

Dey

Marbaniang

, et al Grape seed extract: having a potential health benefits. J Food Sci Technol. 2020;57:1205-1215.

13.

Topalović

Knežević

Bajagić

, et al Chapter 20 - grape (Vitis vinifera L.): health benefits and effects of growing conditions on quality parameters. In: Ozturk

Egamberdieva

Pešić

, eds. Biodiversity and Biomedicine. Academic Press; 2020;385-401.

14.

VanWagenen

Larsen

Cardellina

, et al Ulosantoin, a potent insecticide from the sponge Ulosa ruetzleri. J Org Chem. 1993;58:335-337.

15.

Harborne

AJ.

Phytochemical Methods a Guide to Modern Techniques of Plant Analysis. springer science & business media; 1998.

16.

Sultana

Azad

Rahman

Muhit

Rahman

SA.

Phytochemical and biological investigation of Stevia rebaudiana (Bert.) leaves grown in Bangladesh. Dhaka U J Pharm Sci. 2020;19:191-197.

17.

Hoque

Khan

Rashid

, et al Antimicrobial, antioxidant, and cytotoxic properties of endophytic fungi isolated from Thysanolaena maxima Roxb., Dracaena spicata Roxb. and Aglaonema hookerianum Schott. BMC Complement Med Ther. 2023;23:347.

18.

Salve

Vinchurkar

Raut

, et al. An evaluation of antimicrobial, anticancer, anti-Inflammatory and antioxidant activities of silver nanoparticles synthesized from leaf extract of Madhuca longifolia utilizing quantitative and qualitative methods. Molecules. 2022;27(19):6404.

19.

Rashid

Hossain

Zahan

, et al Chemico-pharmacological and computational studies of Ophiorrhiza fasciculata D. Don and Psychotria silhetensis Hook. f. focusing cytotoxic, thrombolytic, anti-inflammatory, antioxidant, and antibacterial properties. Heliyon. 2023;9:e20100.

20.

Bulbul

Hossain

Haque

, et al Two rare flavonoid glycosides from Litsea glutinosa (Lour.) C. B. Rob.: experimental and computational approaches endorse antidiabetic potentiality. BMC Complement Med Ther. 2024;24:69.

21.

Ezeja

Omeh

Ezeigbo

Ekechukwu

Evaluation of the analgesic activity of the methanolic stem bark extract of Dialium guineense (Wild). Ann Med Health Sci Res. 2011;1:55-62.

22.

Rahman

Soma

Sultana

, et al Exploring therapeutic potential of Woodfordia fruticosa (L.) Kurz leaf and bark focusing on antioxidant, antithrombotic, antimicrobial, anti-inflammatory, analgesic, and antidiarrheal properties. Heal Sci Rep. 2023;6:e1654.

23.

Farzana

Hossain

El-Shehawi

, et al Phenolic constituents from Wendlandia tinctoria var. grandis (Roxb.) DC. Stem deciphering pharmacological potentials against oxidation, hyperglycemia, and diarrhea: phyto-pharmacological and computational approaches. Molecules. 2022;27:5957.

24.

Goswami

Sen

Possible antineoplastic agents, part 15: synthesis, biological activity and quantitative structure activity relationship of substituted-2-(4’-methoxybenzenesulphonamido) glutaric acid analogs against Ehrlich ascites carcinoma. Pharm. 2001;56:366-371.

25.

Gopal Pandit

Honganoor Puttananjaiah

Harohally

Appasaheb Dhale

Functional attributes of a new molecule-2-hydroxymethyl-benzoic acid 2’-hydroxy-tetradecyl ester isolated from Talaromyces purpureogenus CFRM02. Food Chem. 2018;255:89-96.

26.

Cerny

The aroma side of the Maillard reaction. Ann N Y Acad Sci. 2008;1126:66-71.

27.

Tranylcypromine. In: LiverTox: Clinical and Research Information on Drug-Induced Liver Injury. National Institute of Diabetes and Digestive and Kidney Diseases; 2012. Accessed April 17, 2023. http://www.ncbi.nlm.nih.gov/books/NBK548572/

28.

Ogata

Matsumoto

Kida

, et al Synthesis and antifungal activity of a series of novel 1,2-disubstituted propenones. J Med Chem. 1987;30:1497-1502.

29.

Han

Sun

Chen

Antimicrobial susceptibility and antibacterial mechanism of limonene against Listeria monocytogenes. Molecules. 2019;25:33.

30.

Kim

Zou

Kim

, et al Selective peroxynitrite scavenging activity of 3-methyl-1,2-cyclopentanedione from coffee extract. J Pharm Pharmacol. 2002;54:1385-1392.

31.

Almahboub

Narancic

Devocelle

, et al Biosynthesis of 2-aminooctanoic acid and its use to terminally modify a lactoferricin B peptide derivative for improved antimicrobial activity. Appl Microbiol Biotechnol. 2018;102:789-799.

32.

Sung

Jung

Lee

Kim

Lee

Antimicrobial Effect of Furaneol Against Human Pathogenic Bacteria and Fungi. J Microbiol Biotechnol. Published online 2006. Accessed April 17, 2023.https://scholar.google.com/scholar_lookup?title=Antimicrobial+Effect+of+Furaneol+Against+Human+Pathogenic+Bacteria+and+Fungi&author=Sung%2C+W.S.+%28Kyungpook+National+University%2C+Daegu%2C+Republic+of+Korea%29&publication_year=2006

33.

Yale

Pribyl

Braker

Bernstein

Lott

WA.

Muscle-relaxing compounds similar to 3-(o-toloxy)-1,2-propanediol. II. Substituted alkanediols. J Am Chem Soc. 1950;72:3716-3718.

34.

Kumar

Koh

Kim

Gupta

Dutta

PK.

A new chitosan-thymine conjugate: synthesis, characterization and biological activity. Int J Biol Macromol. 2012;50:493-502.

35.

Rubab

Chelliah

Saravanakumar

, et al Bioactive potential of 2-methoxy-4-vinylphenol and benzofuran from Brassica oleracea L. var. capitate f, rubra (red cabbage) on oxidative and microbiological stability of beef meat. Foods. 2020;9:568.

36.

Ashraf

Zubair

Rizwan

, et al Chemical composition, antioxidant and antimicrobial potential of essential oils from different parts of Daphne mucronata Royle. Chem Cent J. 2018;12:135.

37.

Singh

Mansoori

Jiwani

, et al Antioxidant and antimicrobial study of Schefflera vinosa leaves crude extracts against rice pathogens. Arab J Chem. 2021;14:103243.

38.

Zhang

Robertson

Kolachana

Davison

Smith

MT.

Benzene metabolite, 1,2,4-benzenetriol, induces micronuclei and oxidative DNA damage in human lymphocytes and HL60 cells. Environ Mol Mutagen. 1993;21:339-348.

39.

Kadiri

Sevugapperumal

Nallusamy

, et al Pan-genome analysis and molecular docking unveil the biocontrol potential of Bacillus velezensis VB7 against Phytophthora infestans. Microbiol Res. 2023;268:127277.

40.

Junior

Pastore

Limonene and its oxyfunctionalized compounds: biotransformation by microorganisms and their role as functional bioactive compounds. Food Sci Biotechnol. 2009;18:833-841.

41.

Hiramoto

Nasuhara

Michikoshi

Kato

Kikugawa

DNA strand-breaking activity and mutagenicity of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP), a Maillard reaction product of glucose and glycine. Mutat Res Toxicol Env Mutagen. 1997;395:47-56.

42.

Boubakri

Al-Ayed

Mansour

, et al Bioactive NHC-derived palladium complexes: synthesis, catalytic activity for the Suzuki-Miyaura coupling of aryl chlorides and bromides and their antibacterial activities. J Coord Chem. 2019;72:2688-2704.

43.

Vijesh

Isloor

Shetty

Sundershan

Fun

HK.

New pyrazole derivatives containing 1,2,4-triazoles and benzoxazoles as potent antimicrobial and analgesic agents. Eur J Med Chem. 2013;62:410-415.

44.

Kocaçalışkan

Talan

Terzi

Antimicrobial activity of catechol and pyrogallol as allelochemicals. Z Civilistische -forsch C. 2006;61:639-642.

45.

Xia

Cao

Tan

, et al Chemical composition and fungicidal activity of Murraya microphylla Essential oOil against Colletotrichum gloeosporioides. J Essent Oil Bearing Plants. 2020;23:678-685.

46.

Aberchane

Satrani

Fechtal

Chaouch

Effet de l’infection du bois de Cèdre de l’Atlas par Trametes pini et Ungulina officinalis sur la composition chimique et l’activité antibactérienne et antifongique des huiles essentielles. Acta Bot Gallica. 2003;150:223-229.

47.

Qiu

Lin

Chen

Zhang

5-Hydroxymethylfurfural exerts negative effects on gastric mucosal epithelial cells by inducing oxidative sStress, apoptosis, and tight junction disruption. J Agric Food Chem. 2022;70:3852-3861.

48.

Lacerda-Neto

Barbosa

Quintans-Junior

Coutinho

da Cunha

FA.

The complex pharmacology of natural products. Future Med Chem. 2019;11:797-799.

49.

Wang

Zhu

Larvicidal activity of Zanthoxylum acanthopodium essential oil against the malaria mosquitoes, Anopheles anthropophagus and Anopheles sinensis. Malar J. 2018;17:194.

50.

Aggarwal

Khanuja

SPS

Ahmad

, et al Antimicrobial activity profiles of the two enantiomers of limonene and carvone isolated from the oils of Mentha spicata and Anethum sowa. Flavour Fragrance J. 2002;17:59-63.

51.

Ito

Hirose

Imaida

Antioxidants: Carcinogenic and Chemopreventive Properties. In: Bertino

, ed. Encyclopedia of Cancer. 2nd ed. Academic Press; 2002;89-101.

52.

Ibibia

Olabisi

Oluwagbemiga

Gas chromatography-mass spectrometric analysis of methanolic leaf extracts of Lannea kerstingii and Nauclea diderrichii, two medicinal plants used for the treatment of gastrointestinal tract infections. Gas. 2016;9:179-182.

53.

Aoki

Uchida

Enhanced Formation of 3-(methyithio)-1-propanol in a salt-tolerant yeast, Zygosaccharomyces rouxii, due to deficiency of S-A denosylmethionine synthase. Agric Biol Chem. 1991;55:2113-2116.

54.

Yuan

Chen

Conjugated linolenic acids and their bioactivities: a review. Food Funct. 2014;5:1360-1368.

55.

Cynthia

Hery

Akhmad

Antibacterial and antioxidant activities of pyrogallol and synthetic pyrogallol dimer. Res J Chem Env. 2018;22:39-47.

56.

Davidson

den Boer

Ritmeijer

Paromomycin. Trans R Soc Trop Med Hyg. 2009;103:653-660.

57.

Izzat

Bennett

EO.

The Potentiation of the Antimicrobial Activities of Cutting Fluid Preservatives by EDTA. Univ. of Houston, TX; 1978.

58.

Yellu

Bhukya

Evaluation of anticancer activity of methanolic extract of Hiptage benghalensis (L.) Kurz on cancer cell lines. Pharmacogn Res. 2018;10:309.

59.

Srivastava

Mukerjee

Verma

GC-MS analysis of phytocomponents in, pet ether fraction of Wrightia tinctoria seed. Pharmacogn J. 2015;7:249-253. Published online January 1, 2015. doi:10.5530/pj.2015.4.7

60.

Matsubara

Kinoshita

Koyama

, et al Anti-tyrosinase activity of lichen metabolites and their synthetic analogues. J Hattori Bot Lab. 1997;83:179-185.

61.

Rajalakshmi

Mohan

GC-MS Analysis of Bioactive Components of Myxopyrum serratulum A.W. Hill (Oleaceae). Int J Pharm Sci. 2016;38:30-35.

62.

Lee

Purse

Pryce

Horgan

Wareing

PF.

Dihydroconiferyl alcohol - a cell division factor from Acer species. Planta. 1981;152:571-577.

63.

Fagali

Catalá

Antioxidant activity of conjugated linoleic acid isomers, linoleic acid and its methyl ester determined by photoemission and DPPH techniques. Biophys Chem. 2008;137:56-62.

64.

Paudel

Pant

Cytotoxic activity of crude extracts of Dendrobium amoenum and detection of bioactive compounds by GC-MS. Bot Orient J Plant Sci. 2017;11:38-42.

65.

Zhu

Jiao

, et al Palmitic acid inhibits prostate cancer cell proliferation and metastasis by suppressing the PI3K/Akt pathway. Life Sci. 2021;286:120046.

66.

Mohammed

Ghaidaa

Imad

HH.

Analysis of bioactive chemical compounds of Nigella sativa using gas chromatography-mass spectrometry. J Pharmacogn Phytother. 2016;8:8-24.

67.

Cheng

Ker

, et al Chemical synthesis of 9(Z)-octadecenamide and its hypolipidemic effect: A bioactive agent found in the essential oil of mountain celery seeds. J Agric Food Chem. 2010;58:1502-1508.

68.

Kim

Karadeniz

Chapter 14 - biological importance and applications of squalene and squalane. In: Kim

ed. Advances in Food and Nutrition Research. Vol. 65. Marine Medicinal Foods. Academic Press; 2012;223-233.

69.

Zhu

Liu

Wang

, et al Anti-BACE1 and antimicrobial activities of steroidal compounds isolated from marine urechis unicinctus. Mar Drugs. 2018;16:94.

70.

Stanner

Weichselbaum

Antioxidants. In: Caballero

, ed. Encyclopedia of Human Nutrition. 3rd ed. Academic Press; 2013;88-99.

71.

Suman

Datta

Chakraborty

, et al Gamma tocotrienol, a potent radioprotector, preferentially upregulates expression of anti-apoptotic genes to promote intestinal cell survival. Food Chem Toxicol. 2013;60:488-496.

72.

Kang

Jang

Mishra

, et al Ergosterol peroxide from Chaga mushroom (Inonotus obliquus) exhibits anti-cancer activity by down-regulation of the β-catenin pathway in colorectal cancer. J Ethnopharmacol. 2015;173:303-312.

73.

Tucker

Townsend

DM.

Alpha-tocopherol: roles in prevention and therapy of human disease. Biomed Pharmacother Bioméd Pharmacother. 2005;59:380-387.

74.

Khalid

Algarni

Homeida

, et al Phytochemical, cytotoxic, and antimicrobial evaluation of Tribulus terrestris L., Typha domingensis Pers., and Ricinus communis L.: scientific evidences for folkloric uses. Evid-Based Complement Altern Med ECAM. 2022;2022:6519712.

75.

Sirikhansaeng

Tanee

Sudmoon

Chaveerach

Major phytochemical as γ-sitosterol disclosing and toxicity testing in Lagerstroemia species. Evid Based Complement Alternat Med. 2017;2017:7209851.

76.

Kavita

Singh

Jha

24-branched Δ5 sterols from Laurencia papillosa red seaweed with antibacterial activity against human pathogenic bacteria. Microbiol Res. 2014;169:301-306.

77.

Alam

Dhar

Hasan

, et al Antidiabetic potential of commonly available fruit plants in Bangladesh: updates on prospective phytochemicals and their reported MoAs. Molecules. 2022;27:8709.

78.

Zeghad

Ahmed

Belkhiri

Heyden

Demeyer

Antioxidant activity of Vitis vinifera, Punica granatum, Citrus aurantium and Opuntia ficus indica fruits cultivated in Algeria. Heliyon. 2019;5:e01575.

79.

Moldovan

Carpa

Fizeșan

, et al Phytochemical profile and biological activities of tendrils and leaves extracts from a variety of Vitis vinifera L. Antioxidants. 2020;9:373.

80.

Lagartoparra

Yhebra

Sardiñas

Buela

LI.

Comparative study of the assay of and the estimate of the medium lethal dose (LD50 value) in mice, to determine oral acute toxicity of plant extracts. Phytomedicine. 2001;8:395-400.

81.

Meyer

Ferrigni

Putnam

, et al Brine shrimp: a convenient general bioassay for active plant constituents. Planta Med. 1982;45:31-34.

82.

Jannat

Hossain

El-Shehawi

, et al Chemical and pharmacological profiling of Wrightia coccinea (roxb. Ex hornem.) sims focusing antioxidant, cytotoxic, antidiarrheal, hypoglycemic, and analgesic properties. Molecules. 2022;27:4024.

83.

Harbeoui

Hichami

Wannes

, et al Anti-inflammatory effect of grape (Vitis vinifera L.) seed extract through the downregulation of NF-κB and MAPK pathways in LPS-induced RAW264.7 macrophages. S Afr J Bot. 2019;125:1-8.

84.

Giribabu

Karim

Kilari

Kassim

Salleh

Anti-inflammatory, antiapoptotic and proproliferative effects of Vitis vinifera seed ethanolic extract in the liver of streptozotocin-nicotinamide-induced type 2 diabetes in male rats. Can J Diabetes. 2018;42:138-149.

85.

Geronikaki

Gavalas

AM.

Antioxidants and inflammatory disease: synthetic and natural antioxidants with anti-inflammatory activity. Comb Chem High Throughput Screen. 2006;9:425-442.

86.

Schirbel

Reichert

Roll

, et al Impact of pain on health-related quality of life in patients with inflammatory bowel disease. World J Gastroenterol. 2010;16:3168-3177.

87.

Amee

KNS

Hossain

Rohoman

, et al Phytochemical and pharmacological profiling of extracts of Pterygota alata (Roxb.) R. Br. leaves deciphered therapeutic potentialities against pain, hyperglycemia and diarrhea via in vivo approaches. Pharmacol Res Prod. 2024;4:100060.

88.

Nadia

Aicha

Sihem

Abdelmalik

In vivo analgesic activities and safety assessment of Vitis vinifera L and Punica granatum L fruits extracts. Trop J Pharm Res. 2017;16:553.

89.

Aouey

Samet

Fetoui

Simmonds

MSJ

Bouaziz

Anti-oxidant, anti-inflammatory, analgesic and antipyretic activities of grapevine leaf extract (Vitis vinifera) in mice and identification of its active constituents by LC-MS/MS analyses. Biomed Pharmacother. 2016;84:1088-1098.

90.

Laube

Kniess

Pietzsch

Development of antioxidant COX-2 inhibitors as radioprotective agents for radiation therapy-a hypothesis-driven review. Antioxidants. 2016;5:14.

91.

Kaur

Singh

Kumar

Comparative antidiarrheal and antiulcer effect of the aqueous and ethanolic stem bark extracts of Tinospora cordifolia in rats. J Adv Pharm Technol Res. 2014;5:122-128.

92.

Agunu

Yusuf

Andrew

Zezi

Abdurahman

EM.

Evaluation of five medicinal plants used in diarrhoea treatment in Nigeria. J Ethnopharmacol. 2005;101:27-30.

93.

Gao

Ling

Liu

CX.

Antidiarrhoeal and intestinal modulatory activities of Wei-chang-an-wan extract. J Ethnopharmacol. 2009;125:450-455.

94.

Zewdie

Bhoumik

Wondafrash

Tuem

KB.

Evaluation of in-vivo antidiarrhoeal and in-vitro antibacterial activities of the root extract of Brucea antidysenterica J. F. Mill (Simaroubaceae). BMC Complement Med Ther. 2020;20:201-211.

95.

Panganamala

Miller

Gwebu

Sharma

Cornwell

DG.

Differential inhibitory effects of vitamin E and other antioxidants on prostaglandin synthetase, platelet aggregation and lipoxidase. Prostaglandins. 1977;14:261-271.

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