# Peelu (Salvadora oleoides Decne.): An Unexplored Medicinal Fruit with Minerals, Antioxidants, and Phytochemicals

Authors: Kashif Razzaq; Muhammad Muzzammal Sadiq; Hashir Ashraf; Ambreen Naz; Abid Hussain; Amir Maqbool; Muhammad Tanveer Altaf; Sami Ullah; Gulzar Akhtar; Hafiz Nazar Faried; Muhammad Amin; Ishtiaq Ahmad Rajwana; Ahmad Sattar Khan; Saleh Alfarraj; Mohammad Javed Ansari; Ammara Saleem
Journal: BioMed Research International
 (2022)

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## Abstract

The Peelu (Salvadora oleoides Decne.) fruit is well known for its nutritional and medicinal values. The current study analyzed the chemical composition of Salvadora oleoides fruit. Fresh Peelu fruits were harvested, and physicochemical properties, proximate composition, macro- and micronutrients, and phytochemical properties were determined. Moreover, ethanol and methanol fruit extract was analyzed for physicochemical properties. The Peelu fruit seemed to be a potential source of essential macro- ((nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg)) and micronutrients (zinc (Zn), manganese (Mn), iron (Fe), and copper (Cu)). The fruit had significant biochemical properties (total soluble solids (TSS), total acidity (TA), and TSS : TA ratio) with appreciable moisture, crude fiber, and ash contents. The fruit extracts demonstrated significantly higher antioxidants and phenolics, ascorbic acid contents, and carotenoids. Phytochemical screening of fruit revealed the presence of coumarins, flavonoids, phlobatannins, tannins, and terpenoids. Physicochemical and sensory evaluation of extracts indicated its potential for further in vivo study trials. The Peelu fruit was found to be a good source of mineral nutrients, proximate contents, vitamins (ascorbic acid and carotenoid), phytochemicals (total phenolic sand antioxidant contents), and pharmaceutically important metabolites that can be used as functional drink.


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## Body

## 1. Introduction

Peelu (Salvadora oleoides Decne.) is a member of the Salvedoraceae family comprising of three genera, i.e., Azima, Dobera, and Salvadora. Ten (10) species of this family are mainly distributed in tropical and subtropical regions of Africa and Asia [1]. In Pakistan, the family is represented by a single genus Salvadora, with two species, i.e., S. persica and S. oleoides [2]. Recently a new species S. alii has also been reported and described from Sindh province of the country [3, 4]. The Salvadora species can grow in saline, arid, and semiarid regions [5] and serve as an excellent source of food for camels and goats. Moreover, the effective plantations can be used as shelterbelts and windbreaks [6].It is considered a shrub or tree, 4-10 m in height with a twisted trunk. The plant had deep root system and usually known as xerophytes and facultative halophytes with high tolerance to salinity [2]. Leaves are simple, small, and high in number. It is a cross-pollinated plant [4] having greenish-white and greenish-yellow flowers born in leaves axil [6]. The fruit is a small, globose, smooth drupe with red yellow or purple color. The fruits ripen during May and June [6]. The fruit is sweet and peppery in taste with a pungent smell and eaten when fully ripe. Seed oil is pale green that is not fit for consumption [7]. Several therapeutic and industrial uses are well known about this plant which encourage the scientist in exploring more information about this medicinal plant [4]. The plant is well known for its antiulcer, antifungal, antiparasitic, antiviral, and antibacterial properties [8–10]. Leaves are used in the treatment of open wounds and act as a blood purifier and cooling agent. The stem is used to cure fever, asthma, cough, leprosy, rheumatism, and anthelmintic with diuretic properties [5]. Roots are effective against chest and teeth diseases [11, 12]. The oil can be used to manufacture candles, while oil cake is used as feed for animals [13].Apart from the medicinal and nutritional importance, Peelu is a wild ignored crop with a high commercial potential for consumption and value addition. The prevalence of this plant is decreasing swiftly which may cause its extinction in the near future. Therefore, this valuable species could be conserved through exploring and promoting its commercial value. The current study examined the nutritional status of Peelu fruits. The results of the study would improve the understanding and importance of the nutritional value of the plant.


## 2. Materials and Methods

The Peelu fruits were harvested from a commercial field (29°06′12.64″ N 70°19′30.14″ E) by considering size uniformity and free from disease infestation. The characterization of nutritional composition was performed at Postharvest Laboratory, MNS-University of Agriculture, Multan, Pakistan under 30±5°C temperature and 60-65% and relative humidity. Furthermore, proximate composition, mineral quantification, and phytochemical composition were explored. Hundred (100) grams of Peelu fruits were used as an experimental unit and replicated four times.


### 2.1. Compositional Analysis

#### 2.1.1. Moisture (%)

The moisture contents of the Peelu fruit were determined by method no. 44-15 according to AACC [14]. The sample was dried in a hot air oven at 105±5°C till constant weight. The mathematical expression to compute the moisture percentage is given in
(1)Moisture%=Freshweightg−OvendriedweightgFreshweightg×100.


#### 2.1.2. Crude Fat (%)

Crude fat was determined by the Soxhlet apparatus. The 2 g sample was placed in a paper thimble after removing moisture. Fat extraction was done with 75 to 100 mL diethyl ether or hexane. The extraction procedure was repeated four times with solvent. The heat was turned off after the completion of washing. The sample was placed in a hot air oven for the removal of solvent residues followed by cooling in a desiccator. The percentage of crude fat was computed according to(2)Crudefat%=Sampleweightwithfat−SampleweightwithoutfatgSampleweightg×100.


#### 2.1.3. Crude Protein (%)

The crude protein was estimated by the Kjeldahl method. A 5 g sample was added in a preweighed digestion flask to which 3 g digestion mixture containing copper sulfate (CuSO4), potassium sulfate (K2SO4), and ferrous sulfate (Fe2SO4) at the ratio of 5 : 94 : 1, and 25-30 mL sulfuric acid was added. The flask was placed on a burner and heated until 2-3 mL of the solution was left. The sample was then diluted up to 50 mL. Afterwards, 10 mL of the sample and 10 mL of NaOH (40%) were added in the reaction flask of the Kjeldahl apparatus. The 10 mL of boric acid (4%) was added to the receiving beaker of the Kjeldahl apparatus. The distillation continued until the volume of boric acid reached 20-22 mL and turned yellow. It was titrated with 0.1 N H2SO4 and N was computed according to equation (3). Form the determined N percentage, crude protein was calculated according to
(3)N%=0.0014×volumeof0.1Nsulfuricacid×VolumeofdilutionWeightofsample×Volumeofsampletaken×100,Crudeprotein%=N%×6.25.


#### 2.1.4. Crude Fiber (%)

Crude fiber was determined in four steps. Primarily, a fat-free sample was added with 200 mL of boiling sulfuric acid (1.25%) till simmering (70-80°C). Afterwards, the sample was filtered with a linen cloth in the fluted funnel. In the second step, the acid-treated sample was subjected to alkaline medium of NaOH (1.25%) followed by heating, simmering, and filtration. In the third step, the treated sample was subjected to moisture removal according to the standard protocol. The weight of the sample obtained after moisture removal was denoted asW1. Finally, ashing was carried out. The weight of the sample after ashing was designated as W2. The cured fiber was calculated according to
(4)Crudefiber%=W1−W2Weightofsample×100.


#### 2.1.5. Ash (%)

The ash (%) content was estimated by Muffle Furnace (MF-1/02, PCSIR, Pakistan). The 3 g dry sample was incinerated after charring in Muffle Furnace according to Method 08-01 AACC [14] at 550°C till grayish-white residue. The ash content was computed by using
(5)Ash%=WeightofashgWeightofsampleg×100.


### 2.2. Mineral Profiling

The fresh fruits were washed with distilled water and dried in a hot air oven at 105°C till constant weight. The dried fruits were ground proceeded by acid digestion. The resultant solution was used to determine various mineral nutrients as nitrogen (N) by the micro-Kjeldahl method, phosphorus (P) by vanadomolybdo analysis, and potassium (K) by flame photometer as elaborated by Ullah et al. [15]. Additionally, other elements as Zn, Ca, Mn, and Fe were estimated by an Atomic Absorption Spectrophotometer (2-8200 Series Polarized Zeeman, Hitachi, Japan) by using a hollow cathode lamp [15].


### 2.3. Phenolic and Antioxidants

The ascorbic acid was determined by following the protocol of Kumar et al. [4] using trichloroacetic acid as reference. Moreover, ascorbic acid contents were calculated using the standard curve of L-ascorbic acid and expressed as μg·100 g-1·DW. Total carotenoids were determined and expressed as μg·g-1 of β-carotene equivalent [16]. Total phenolic contents (TPC) were estimated by following Folin–Ciocalteu (FC) method as discussed by Razzaq et al. [17]. The concentration of TPC was showed as mg GAE·100 g-1. The antioxidant activity was measured through 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) by spectrophotometer in ELX800 Microplate Reader (Bio-Tek Instruments, Inc., Winooski, VT, USA) as outlined by Mimica-Dukić et al. [18] and was expressed as %. The inhibition percentage was computed according to
(6)Inhibition%=AB−AAAB×100,where AB is the absorption of a blank sample and AA is the absorbance of the tested sample.


### 2.4. Tannins, Coumarins, Phlobatannins, Flavonoids, and Terpenoids

The fruit pulp (50 mg) was boiled in distilled water (20 mL) followed by filtration. After boiling, FeCl3 (0.1%) was added to the filtrate and a color change was noticed as blue-black or brownish-green as evidence of tannin presence. For the determination of coumarins, a fruit sample (300 g) was added to a test tube covered with a filter paper soaked in NaOH (1 N). The sample containing the test tube was placed in a hot water bath. The filter paper was removed and the sample was observed under UV light. The appearance of a yellow color indicated the presence of coumarins. In a small test tube, 300 mg of fruit was covered with filter paper moistened with 1 N NaOH. The test tube was placed in a boiling water bath for few minutes. After removing the filter paper, it was examined under UV light, yellow fluorescence indicated the presence of coumarins. The Peelu fruit was boiled in diluted hydrochloric acid (1%). The resultant deposition of red-colored precipitates indicated the occurrence of phlobatannins. The fruit sample (50 mg) was added in 100 mL of distilled water to obtain a filtrate. In the prepared filtrate, 5 mL of dilute ammonia solution plus few drops of concentrated H2SO4. The flavonoids were confirmed by the appearance of the yellow color [19]. The determination of terpenoids was done by taking 5 mL of fruit pulp tissues with a dilution of 1 mg mL-1, mixed with 2 mL chloroform and 3 mL of concentrated H2SO4. The reddish-brown color indicated the presence of terpenoids as described by Saeed et al. [19].


### 2.5. Biochemical Analysis

The total soluble solids (TSS) were measured by a digital refractometer (ATAGO, RX5000, Atago Co. Ltd., Itabashi-ku, Tokyo, Japan) and expressed as °Brix. The total acidity (TA) was assessed via the acid-base titration method. The juice sample was titrated against NaOH (0.1 N) by using a phenolphthalein indicator. The pH of juice was estimated through a digital pH meter (Starter 3100 OHAUS Corporation, USA).


### 2.6. Functional/Nutraceutical Drink

Functional drink (Nutra-1) was developed by using various ingredients include citric acid, carboxymethylcellulose (CMC), aspartame, sodium benzoate, flavor, and food-grade color along with propolis extract of following treatments at the dose rate of 400 mg/500 mL of the functional drink. The treatment planning of drinks was designed asT0 (placebo), T1 (functional drink with ethanol extract of Peelu), and T2 (functional drink with methanol of extract of Peelu).


### 2.7. Functional Drink Analysis

The functional drinks were analyzed for their physicochemical attributes including TSS, TA, and pH. Sensory evaluation (color, flavor, sweetness, sourness, and overall acceptability) was done by using the 9-point hedonic scale (9 = extremely like, 1 = extremely dislike) as specified by Meilgaard et al. [20] during 10 days intervals of storage period (30 days).


### 2.8. Statistical Analysis

The experiment was carried out according to completely randomized design (CRD) with four replications. The data regarding different studied attributes were analyzed by descriptive statistics and presented. Statistix® statistical software was used for the data analysis [21]. The data regarding nutritional composition of functional drinks were analyzed by one-way analysis of variance (ANOVA). The normality in the data was tested indicating that the data had normal distribution. Therefore, the original data were used in the analysis. The least significant difference test at 95% probability was used to compare the means where ANOVA indicated significant differences.


## 2.1. Compositional Analysis

### 2.1.1. Moisture (%)

The moisture contents of the Peelu fruit were determined by method no. 44-15 according to AACC [14]. The sample was dried in a hot air oven at 105±5°C till constant weight. The mathematical expression to compute the moisture percentage is given in
(1)Moisture%=Freshweightg−OvendriedweightgFreshweightg×100.


### 2.1.2. Crude Fat (%)

Crude fat was determined by the Soxhlet apparatus. The 2 g sample was placed in a paper thimble after removing moisture. Fat extraction was done with 75 to 100 mL diethyl ether or hexane. The extraction procedure was repeated four times with solvent. The heat was turned off after the completion of washing. The sample was placed in a hot air oven for the removal of solvent residues followed by cooling in a desiccator. The percentage of crude fat was computed according to(2)Crudefat%=Sampleweightwithfat−SampleweightwithoutfatgSampleweightg×100.


### 2.1.3. Crude Protein (%)

The crude protein was estimated by the Kjeldahl method. A 5 g sample was added in a preweighed digestion flask to which 3 g digestion mixture containing copper sulfate (CuSO4), potassium sulfate (K2SO4), and ferrous sulfate (Fe2SO4) at the ratio of 5 : 94 : 1, and 25-30 mL sulfuric acid was added. The flask was placed on a burner and heated until 2-3 mL of the solution was left. The sample was then diluted up to 50 mL. Afterwards, 10 mL of the sample and 10 mL of NaOH (40%) were added in the reaction flask of the Kjeldahl apparatus. The 10 mL of boric acid (4%) was added to the receiving beaker of the Kjeldahl apparatus. The distillation continued until the volume of boric acid reached 20-22 mL and turned yellow. It was titrated with 0.1 N H2SO4 and N was computed according to equation (3). Form the determined N percentage, crude protein was calculated according to
(3)N%=0.0014×volumeof0.1Nsulfuricacid×VolumeofdilutionWeightofsample×Volumeofsampletaken×100,Crudeprotein%=N%×6.25.


### 2.1.4. Crude Fiber (%)

Crude fiber was determined in four steps. Primarily, a fat-free sample was added with 200 mL of boiling sulfuric acid (1.25%) till simmering (70-80°C). Afterwards, the sample was filtered with a linen cloth in the fluted funnel. In the second step, the acid-treated sample was subjected to alkaline medium of NaOH (1.25%) followed by heating, simmering, and filtration. In the third step, the treated sample was subjected to moisture removal according to the standard protocol. The weight of the sample obtained after moisture removal was denoted asW1. Finally, ashing was carried out. The weight of the sample after ashing was designated as W2. The cured fiber was calculated according to
(4)Crudefiber%=W1−W2Weightofsample×100.


### 2.1.5. Ash (%)

The ash (%) content was estimated by Muffle Furnace (MF-1/02, PCSIR, Pakistan). The 3 g dry sample was incinerated after charring in Muffle Furnace according to Method 08-01 AACC [14] at 550°C till grayish-white residue. The ash content was computed by using
(5)Ash%=WeightofashgWeightofsampleg×100.


## 2.1.1. Moisture (%)

The moisture contents of the Peelu fruit were determined by method no. 44-15 according to AACC [14]. The sample was dried in a hot air oven at 105±5°C till constant weight. The mathematical expression to compute the moisture percentage is given in
(1)Moisture%=Freshweightg−OvendriedweightgFreshweightg×100.


## 2.1.2. Crude Fat (%)

Crude fat was determined by the Soxhlet apparatus. The 2 g sample was placed in a paper thimble after removing moisture. Fat extraction was done with 75 to 100 mL diethyl ether or hexane. The extraction procedure was repeated four times with solvent. The heat was turned off after the completion of washing. The sample was placed in a hot air oven for the removal of solvent residues followed by cooling in a desiccator. The percentage of crude fat was computed according to(2)Crudefat%=Sampleweightwithfat−SampleweightwithoutfatgSampleweightg×100.


## 2.1.3. Crude Protein (%)

The crude protein was estimated by the Kjeldahl method. A 5 g sample was added in a preweighed digestion flask to which 3 g digestion mixture containing copper sulfate (CuSO4), potassium sulfate (K2SO4), and ferrous sulfate (Fe2SO4) at the ratio of 5 : 94 : 1, and 25-30 mL sulfuric acid was added. The flask was placed on a burner and heated until 2-3 mL of the solution was left. The sample was then diluted up to 50 mL. Afterwards, 10 mL of the sample and 10 mL of NaOH (40%) were added in the reaction flask of the Kjeldahl apparatus. The 10 mL of boric acid (4%) was added to the receiving beaker of the Kjeldahl apparatus. The distillation continued until the volume of boric acid reached 20-22 mL and turned yellow. It was titrated with 0.1 N H2SO4 and N was computed according to equation (3). Form the determined N percentage, crude protein was calculated according to
(3)N%=0.0014×volumeof0.1Nsulfuricacid×VolumeofdilutionWeightofsample×Volumeofsampletaken×100,Crudeprotein%=N%×6.25.


## 2.1.4. Crude Fiber (%)

Crude fiber was determined in four steps. Primarily, a fat-free sample was added with 200 mL of boiling sulfuric acid (1.25%) till simmering (70-80°C). Afterwards, the sample was filtered with a linen cloth in the fluted funnel. In the second step, the acid-treated sample was subjected to alkaline medium of NaOH (1.25%) followed by heating, simmering, and filtration. In the third step, the treated sample was subjected to moisture removal according to the standard protocol. The weight of the sample obtained after moisture removal was denoted asW1. Finally, ashing was carried out. The weight of the sample after ashing was designated as W2. The cured fiber was calculated according to
(4)Crudefiber%=W1−W2Weightofsample×100.


## 2.1.5. Ash (%)

The ash (%) content was estimated by Muffle Furnace (MF-1/02, PCSIR, Pakistan). The 3 g dry sample was incinerated after charring in Muffle Furnace according to Method 08-01 AACC [14] at 550°C till grayish-white residue. The ash content was computed by using
(5)Ash%=WeightofashgWeightofsampleg×100.


## 2.2. Mineral Profiling

The fresh fruits were washed with distilled water and dried in a hot air oven at 105°C till constant weight. The dried fruits were ground proceeded by acid digestion. The resultant solution was used to determine various mineral nutrients as nitrogen (N) by the micro-Kjeldahl method, phosphorus (P) by vanadomolybdo analysis, and potassium (K) by flame photometer as elaborated by Ullah et al. [15]. Additionally, other elements as Zn, Ca, Mn, and Fe were estimated by an Atomic Absorption Spectrophotometer (2-8200 Series Polarized Zeeman, Hitachi, Japan) by using a hollow cathode lamp [15].


## 2.3. Phenolic and Antioxidants

The ascorbic acid was determined by following the protocol of Kumar et al. [4] using trichloroacetic acid as reference. Moreover, ascorbic acid contents were calculated using the standard curve of L-ascorbic acid and expressed as μg·100 g-1·DW. Total carotenoids were determined and expressed as μg·g-1 of β-carotene equivalent [16]. Total phenolic contents (TPC) were estimated by following Folin–Ciocalteu (FC) method as discussed by Razzaq et al. [17]. The concentration of TPC was showed as mg GAE·100 g-1. The antioxidant activity was measured through 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) by spectrophotometer in ELX800 Microplate Reader (Bio-Tek Instruments, Inc., Winooski, VT, USA) as outlined by Mimica-Dukić et al. [18] and was expressed as %. The inhibition percentage was computed according to
(6)Inhibition%=AB−AAAB×100,where AB is the absorption of a blank sample and AA is the absorbance of the tested sample.


## 2.4. Tannins, Coumarins, Phlobatannins, Flavonoids, and Terpenoids

The fruit pulp (50 mg) was boiled in distilled water (20 mL) followed by filtration. After boiling, FeCl3 (0.1%) was added to the filtrate and a color change was noticed as blue-black or brownish-green as evidence of tannin presence. For the determination of coumarins, a fruit sample (300 g) was added to a test tube covered with a filter paper soaked in NaOH (1 N). The sample containing the test tube was placed in a hot water bath. The filter paper was removed and the sample was observed under UV light. The appearance of a yellow color indicated the presence of coumarins. In a small test tube, 300 mg of fruit was covered with filter paper moistened with 1 N NaOH. The test tube was placed in a boiling water bath for few minutes. After removing the filter paper, it was examined under UV light, yellow fluorescence indicated the presence of coumarins. The Peelu fruit was boiled in diluted hydrochloric acid (1%). The resultant deposition of red-colored precipitates indicated the occurrence of phlobatannins. The fruit sample (50 mg) was added in 100 mL of distilled water to obtain a filtrate. In the prepared filtrate, 5 mL of dilute ammonia solution plus few drops of concentrated H2SO4. The flavonoids were confirmed by the appearance of the yellow color [19]. The determination of terpenoids was done by taking 5 mL of fruit pulp tissues with a dilution of 1 mg mL-1, mixed with 2 mL chloroform and 3 mL of concentrated H2SO4. The reddish-brown color indicated the presence of terpenoids as described by Saeed et al. [19].


## 2.5. Biochemical Analysis

The total soluble solids (TSS) were measured by a digital refractometer (ATAGO, RX5000, Atago Co. Ltd., Itabashi-ku, Tokyo, Japan) and expressed as °Brix. The total acidity (TA) was assessed via the acid-base titration method. The juice sample was titrated against NaOH (0.1 N) by using a phenolphthalein indicator. The pH of juice was estimated through a digital pH meter (Starter 3100 OHAUS Corporation, USA).


## 2.6. Functional/Nutraceutical Drink

Functional drink (Nutra-1) was developed by using various ingredients include citric acid, carboxymethylcellulose (CMC), aspartame, sodium benzoate, flavor, and food-grade color along with propolis extract of following treatments at the dose rate of 400 mg/500 mL of the functional drink. The treatment planning of drinks was designed asT0 (placebo), T1 (functional drink with ethanol extract of Peelu), and T2 (functional drink with methanol of extract of Peelu).


## 2.7. Functional Drink Analysis

The functional drinks were analyzed for their physicochemical attributes including TSS, TA, and pH. Sensory evaluation (color, flavor, sweetness, sourness, and overall acceptability) was done by using the 9-point hedonic scale (9 = extremely like, 1 = extremely dislike) as specified by Meilgaard et al. [20] during 10 days intervals of storage period (30 days).


## 2.8. Statistical Analysis

The experiment was carried out according to completely randomized design (CRD) with four replications. The data regarding different studied attributes were analyzed by descriptive statistics and presented. Statistix® statistical software was used for the data analysis [21]. The data regarding nutritional composition of functional drinks were analyzed by one-way analysis of variance (ANOVA). The normality in the data was tested indicating that the data had normal distribution. Therefore, the original data were used in the analysis. The least significant difference test at 95% probability was used to compare the means where ANOVA indicated significant differences.


## 3. Results and Discussion

### 3.1. Compositional Analysis

The compositional profiling of fruit elucidated that it contained significant moisture content (63.2%), crude fat (7.1%), crude protein (4.2%), crude fiber (8.7%), and ash (7.8%) as indicated in Table1. The findings of the current study are closely related to the outcomes of Kumari et al. [22] who documented that ripened fruit of Salvadora persica had significant amounts of moisture (73.8±5.0%), protein (5.9±0.6%), ash (9.3±0.6%), and fiber (10.4±1.9%) which was slightly higher than our results. Variation in proximate composition of the Peelu fruit may be due to different species, climatic conditions, soil type, or growing environment. The studied fruit was harvested from Southern Punjab, Pakistan, which had comparative lesser relative humidity which can be a major contributing factor in the low moisture content of fruit under investigation as compared to the results of Kumari et al. [22].Table 1
Compositional analysis (%) ofSalvadora oleoides fruit.
ParametersMinimumMaximumMean∗Moisture contents54.0771.2363.23±6.3Crude protein2.536.044.23±1.32Crude fat5.359.767.19±2.09Crude fiber4.9513.218.75±3.02Ash5.5312.678.78±2.57∗Mean±SD.


### 3.2. Mineral Profiling

The mineral profiling revealed the presence of a sufficient amount of macronutrients, i.e., nitrogen (N) 209.3 mg·100 g-1·DW, phosphorus (P) 85.0 mg·100 g-1·DW, potassium (K) 631.5 mg·100 g-1·DW, calcium (Ca) 507.5 mg·100 g-1·DW, and magnesium (Mg) 113.7 mg·100 g-1·DW (Table 2). Our results showed that Peelu fruits contain a comparable amount of K and Ca with Dialium guineense [23]. The K is beneficial for cardiovascular disorders by reducing blood pressure by maintaining appropriate K+ and Na+ ratio [24]. Calcium (Ca+2) is a crucial mineral of the human diet that facilitates various biological functions of the body as differentiation, neuronal activities, muscles contraction, and immune responses that leads to apoptosis [25]. The Peelu fruit is rich source of Ca than common fruits, i.e., orange, apple, banana, papaya, chiku, and medicinal plants [13, 26]. The Peelu fruit was found to be a good source of Na+ which is important for various biological processes [27]. The Mg prevents cardiovascular diseases. It maintains bone structure by retaining Ca and vitamin D into bones which prevents osteoporosis [28]. Our results reported optimum concentrations of Mg in the Peelu fruit which are high than nonconventional fruits [29–31]. Hence, the Peelu fruit may be considered for functional foods to meet the daily requirement of minerals.Table 2
Minerals (mg·100 g-1·DW) composition of Salvadora oleoides fruit.
ParametersMaximumMinimumMean∗Nitrogen (N)245.23182.43209.35±22.64Phosphorus (P)101.5665.7885.08±12.62Potassium (K)695.24536.89631.56±60.81Calcium (Ca)589.78422.76507.56±57.68Magnesium (Mg)127.56100.98113.79±11.30Iron (Fe)9123.676758.348068.12±938.53Manganese (Mn)678.9645.78656.84±12.15Zinc (Zn)674.21445.21532.05±97.24Copper (Cu)672.67450.67556.98±88.75∗Mean±SD.The Peelu fruits were rich in many micronutrients like Fe, Mn, Zn, and Cu as 8068.1, 656.2, 532.0, and 556.9 mg·100 g-1·DW, respectively (Table 2). These micronutrients were required for the survival of human health as involved in many biochemical, physiological, and metabolic processes like DNA synthesis, mitotic division, respiration, and healthy aging [32, 33]. Moreover, micronutrient contents of Peelu in this study were at par with Salvadora persica, Ziziphus jujube, and Capparis decidua [29, 30]. However, these were lower than Dialium guineense, Chrysophyllum albidum, and Spondias mombin [30, 31].


### 3.3. Phenolic and Antioxidant Contents

Phenolic substances of different origins and functions can be obtained from plants. Most of these substances have antiviral, anticancerous, and antibacterial properties [34]. Phenolic contents and antioxidant activities of nontraditional fruits plants have limited description in literature [35]. The total phenolic content of the Peelu fruit extract were 598.9±100.5 mg·GAE·100 g-1 (Table 3). These contents are higher from Ficus sycomorus, Diospyros mespiliformis, Parkia biglobosa, and Lannea microcarpa and almost at par with Sclerocarya birrea, Diospyros mespiliformis, and Saba senegalensis and lower than Tamarindus indica and Ziziphus mauritiana [36]. The total phenolic content of the Peelu fruit has been found higher than the red and green peppers and at par with sour cherry and blueberry [23].Table 3
Total phenolic, antioxidant, and ascorbic acid contents ofSalvadora oleoides fruit.
ParametersMinimumMaximumMean∗Total phenolic contents (mg·GAE·100 g-1)525.25675.9598.9±100.35Total antioxidants (% inhibition)60.879.5370.25±7.33Carotenoid (μg·100 g-1·DW)4.678.416.52±1.39Ascorbic acid contents (μg·100 g-1·DW)82.465.673.9±1.15∗Mean±SD.The antioxidants are becoming more important due to their role in maintaining good health and inhibiting disease through scavenging free radicals responsible for the multiplication of many diseases, including cancer, neurodegenerative, and AIDS disorder [22]. The total antioxidant capacity of the Peelu fruit was 70.2±7.3% inhibition (Table 4). Antioxidant contents of the Peelu fruit were higher than banana, water apple, guava, and star fruit [37]. Among the species, the antioxidant contents of S. oleoides were higher than S. persica [22].Table 4
Biochemical attributes ofSalvadora oleoides fruit.
ParametersMinimumMaximumMean∗Total soluble solids (°Brix)7.210.58.9±1.32Titratable acidity (%)0.3250.620.5±0.10Ripening index (ratio)11.232.319.7±7.32pH3.84.13.9±0.11∗Mean±SD.Carotenoids are a good source of antioxidants and considered fat-soluble pigments in plants. Moreover, color development is also dependent on carotenoids synthesis during maturity and ripening of the fruits. The carotenoid content in Peelu were6.5±1.4 μg·100 g-1 (Table 4). Carotenoids obtained from fruits and vegetables significantly reduce the risks of diseases and provide more health benefits [38, 39]; hence, Peelu could be a good choice of carotenoids for dietary intake. Ascorbic acid has antioxidant activity and maintains cellular membrane activity. Ascorbic acid inhibits the conversion of cancer-causing compound N-nitroso from nitrates and nitrides present in fruit and vegetables [40]. In the current study, ascorbic acid contents in the Peelu fruit were 73.9±1.1 μg·100 g-1 (Table 4). Higher ascorbic acid content in Peelu proved its great potential for promoting health benefits. Similarly, a higher amount of ascorbic (67.9 mg·100 g−1·DW) has been reported for S. persica fruit [22].


### 3.4. Biochemical Analysis

Data regarding biochemical profiling of the Peelu fruit indicated that TSS in fruit juice were8.9±1.3°Brix (Table 4). The percent TA was 0.5±0.1 (%) with a pH value of 3.9±0.1. Additionally, the ripening index ratio was 19.7±7.3. These results showed that biochemical attributes of Peelu were at par with other ignored fruits like Averrhoa carambola, Morus alba, and Syzygium cumini [41, 42], comparable with Grewia subinaequalis, and lesser than Prunus persica and Prunus armeniaca [43] suggesting the usage of this rare fruit in different value-added products.


### 3.5. Physicochemical Analysis of Drink

Data regarding various physicochemical attributes (pH and acidity) showed a significant effect of treatments and storage duration on functional drinks prepared from Peelu. However, the interaction between treatments and storage duration was nonsignificant for pH and acidity of the functional drinks. Moreover, a nonsignificant effect of treatment and storage period, as well as their interaction, was recorded for TSS (Figure1).Figure 1
Impact of treatments (T0: placebo; T1: ethanol extract containing functional drink; T2: methanol extract containing functional drink) on overall acceptability of drink during storage period. Vertical bars indicate ±standard error of means. n=3, LSD (P≤0.05) for drinks.The highest concentration of TSS (1.6°Brix) was observed in functional drink with ethanol extract of Peelu as compared to placebo and functional drink with methanol extract of Peelu (1.58°Brix and 1.58°Brix). As far as the storage period was concerned, an increasing trend was observed for TSS for the storage period. The highest concentration of TA (1.67%) was obtained after 30 days of storage period as compared to 0 days (1.5%) (Figure2(a)). An increasing trend has been observed for acidity with prolonging storage period in all treatments. This increase in acidity (%) was more in methanol extract of Peelu when compared with other treatments. After storage of 30 days drink prepared in methanol extract showed 1.03- and 1.06-fold more TA (Figure 2(b)). Among various treatments, functional dinks prepared in methanol extract exhibited the highest (3.48) value for pH. A significant linear decreasing trend (1.13-fold) was observed for the pH of functional drinks with the increase in storage period from day-0 to day-30 (Figure 2(c)).Figure 2
Impact of treatments (T0: placebo; T1: ethanol extract containing functional drink; T2: methanol extract containing functional drink) on total soluble solids (a), acidity (b), and pH (c) of functional drink during the storage period. Vertical bars indicate ±standard error of means. n=3, LSD (P≤0.05) for drinks.Previously, the average TSS documented was about 10.4°Brix in a diet drink of apple prepared with artificial sweetener. Similarly, the results of the current investigation are in corroboration with González-Molina et al. [44], who reported a nonmomentous effect of storage period on TSS of apple and pomegranate juices and their blending. The result of the present study for pH and acidity of functional drinks is similar to Ahmed [45], who observed that a negative correlation exists between acidity and pH of juice during storage. Moreover, these increase and decrease in acidity and pH are closely linked with each other as reported by Klimczak et al. [46]. Similarly, an increase in acidity and a sharp decline in pH was monitored in fruit drink during 90 days of storage [47]. The inverse relationship between pH and acidity has also been observed in yogurt-like beverages including cereals [48]. This increase in acidity may be due to the breakdown of artificial sweeteners as well as citric acid in replacement of sugar for making therapeutic drinks [49]. Additionally, a decrease in pH and increase in acidity may be due to the degradation of polysaccharides and oxidation of reducing sugars that results in the production of acidic components. Furthermore, the development of uronic acid due to the breakdown of pectin may also be the reason for a change in the abovementioned parameters [50].


### 3.6. Sensory Evaluation

Treatments, storage period, and interaction were nonsignificant for sensory parameters of functional drinks. Color of the product/physical appearance is considered a prime factor for selection/rejection of the product by the consumer. With the increase in storage duration color score of the functional drinks was significantly reduced. The color score of the drinks was more at 0-day (6.6), while rescued to 6.1 after 30 days of storage. Among the treatment, the highest color score was obtained fromT2 (6.51±0.07) as compared to T1 and T0 (6.45 and 6.23, respectively) (Figure 3(a)). The flavor of drinks was found higher (7.1±0.06) at 0-day, while the lowest value (6.96) was recorded after 30 days of storage (Figure 3(b)). As far as various treatments are concerned, drink prepared with ethanol extract (T1) showed a better flavor score (7.14), as compared to drinks prepared with methanol extract (7.09) and drink without extract (6.94). Average values documented for sweetness and sourness at 0-day were 7.20 and 7.68, 7.60 and 7.79, and 7.36 and 7.5, respectively, whereas mean values for sweetness and sourness after 30 days of storage were 7.08 and 7.38, 7.22 and 7.5, and 7.18 and 7.39 for T0, T1, and T2 treated drink accordingly. The overall acceptability means a score of functional drinks was recorded about 7.77 during storage.Figure 3
Impact of treatments (T0: placebo; T1: ethanol extract containing functional drink; T2: methanol extract containing functional drink) on color (a), flavor (b), sweetness (c), and sourness (d) of functional drink during storage period. Vertical bars indicate ±standard error of means. n=3, LSD (P≤0.05) for drinks.Our results were in line with Ahmed [45] who reported that sensory evaluation of functional drinks prepared with polyphenol extracts declined in various sensory attributes including color, flavor, sweetness, sourness, and overall acceptability. This may be due to the reason that polyphenols induce color variation because of color imparting bodies linked with the plant polyphenols [51].


## 3.1. Compositional Analysis

The compositional profiling of fruit elucidated that it contained significant moisture content (63.2%), crude fat (7.1%), crude protein (4.2%), crude fiber (8.7%), and ash (7.8%) as indicated in Table1. The findings of the current study are closely related to the outcomes of Kumari et al. [22] who documented that ripened fruit of Salvadora persica had significant amounts of moisture (73.8±5.0%), protein (5.9±0.6%), ash (9.3±0.6%), and fiber (10.4±1.9%) which was slightly higher than our results. Variation in proximate composition of the Peelu fruit may be due to different species, climatic conditions, soil type, or growing environment. The studied fruit was harvested from Southern Punjab, Pakistan, which had comparative lesser relative humidity which can be a major contributing factor in the low moisture content of fruit under investigation as compared to the results of Kumari et al. [22].Table 1
Compositional analysis (%) ofSalvadora oleoides fruit.
ParametersMinimumMaximumMean∗Moisture contents54.0771.2363.23±6.3Crude protein2.536.044.23±1.32Crude fat5.359.767.19±2.09Crude fiber4.9513.218.75±3.02Ash5.5312.678.78±2.57∗Mean±SD.


## 3.2. Mineral Profiling

The mineral profiling revealed the presence of a sufficient amount of macronutrients, i.e., nitrogen (N) 209.3 mg·100 g-1·DW, phosphorus (P) 85.0 mg·100 g-1·DW, potassium (K) 631.5 mg·100 g-1·DW, calcium (Ca) 507.5 mg·100 g-1·DW, and magnesium (Mg) 113.7 mg·100 g-1·DW (Table 2). Our results showed that Peelu fruits contain a comparable amount of K and Ca with Dialium guineense [23]. The K is beneficial for cardiovascular disorders by reducing blood pressure by maintaining appropriate K+ and Na+ ratio [24]. Calcium (Ca+2) is a crucial mineral of the human diet that facilitates various biological functions of the body as differentiation, neuronal activities, muscles contraction, and immune responses that leads to apoptosis [25]. The Peelu fruit is rich source of Ca than common fruits, i.e., orange, apple, banana, papaya, chiku, and medicinal plants [13, 26]. The Peelu fruit was found to be a good source of Na+ which is important for various biological processes [27]. The Mg prevents cardiovascular diseases. It maintains bone structure by retaining Ca and vitamin D into bones which prevents osteoporosis [28]. Our results reported optimum concentrations of Mg in the Peelu fruit which are high than nonconventional fruits [29–31]. Hence, the Peelu fruit may be considered for functional foods to meet the daily requirement of minerals.Table 2
Minerals (mg·100 g-1·DW) composition of Salvadora oleoides fruit.
ParametersMaximumMinimumMean∗Nitrogen (N)245.23182.43209.35±22.64Phosphorus (P)101.5665.7885.08±12.62Potassium (K)695.24536.89631.56±60.81Calcium (Ca)589.78422.76507.56±57.68Magnesium (Mg)127.56100.98113.79±11.30Iron (Fe)9123.676758.348068.12±938.53Manganese (Mn)678.9645.78656.84±12.15Zinc (Zn)674.21445.21532.05±97.24Copper (Cu)672.67450.67556.98±88.75∗Mean±SD.The Peelu fruits were rich in many micronutrients like Fe, Mn, Zn, and Cu as 8068.1, 656.2, 532.0, and 556.9 mg·100 g-1·DW, respectively (Table 2). These micronutrients were required for the survival of human health as involved in many biochemical, physiological, and metabolic processes like DNA synthesis, mitotic division, respiration, and healthy aging [32, 33]. Moreover, micronutrient contents of Peelu in this study were at par with Salvadora persica, Ziziphus jujube, and Capparis decidua [29, 30]. However, these were lower than Dialium guineense, Chrysophyllum albidum, and Spondias mombin [30, 31].


## 3.3. Phenolic and Antioxidant Contents

Phenolic substances of different origins and functions can be obtained from plants. Most of these substances have antiviral, anticancerous, and antibacterial properties [34]. Phenolic contents and antioxidant activities of nontraditional fruits plants have limited description in literature [35]. The total phenolic content of the Peelu fruit extract were 598.9±100.5 mg·GAE·100 g-1 (Table 3). These contents are higher from Ficus sycomorus, Diospyros mespiliformis, Parkia biglobosa, and Lannea microcarpa and almost at par with Sclerocarya birrea, Diospyros mespiliformis, and Saba senegalensis and lower than Tamarindus indica and Ziziphus mauritiana [36]. The total phenolic content of the Peelu fruit has been found higher than the red and green peppers and at par with sour cherry and blueberry [23].Table 3
Total phenolic, antioxidant, and ascorbic acid contents ofSalvadora oleoides fruit.
ParametersMinimumMaximumMean∗Total phenolic contents (mg·GAE·100 g-1)525.25675.9598.9±100.35Total antioxidants (% inhibition)60.879.5370.25±7.33Carotenoid (μg·100 g-1·DW)4.678.416.52±1.39Ascorbic acid contents (μg·100 g-1·DW)82.465.673.9±1.15∗Mean±SD.The antioxidants are becoming more important due to their role in maintaining good health and inhibiting disease through scavenging free radicals responsible for the multiplication of many diseases, including cancer, neurodegenerative, and AIDS disorder [22]. The total antioxidant capacity of the Peelu fruit was 70.2±7.3% inhibition (Table 4). Antioxidant contents of the Peelu fruit were higher than banana, water apple, guava, and star fruit [37]. Among the species, the antioxidant contents of S. oleoides were higher than S. persica [22].Table 4
Biochemical attributes ofSalvadora oleoides fruit.
ParametersMinimumMaximumMean∗Total soluble solids (°Brix)7.210.58.9±1.32Titratable acidity (%)0.3250.620.5±0.10Ripening index (ratio)11.232.319.7±7.32pH3.84.13.9±0.11∗Mean±SD.Carotenoids are a good source of antioxidants and considered fat-soluble pigments in plants. Moreover, color development is also dependent on carotenoids synthesis during maturity and ripening of the fruits. The carotenoid content in Peelu were6.5±1.4 μg·100 g-1 (Table 4). Carotenoids obtained from fruits and vegetables significantly reduce the risks of diseases and provide more health benefits [38, 39]; hence, Peelu could be a good choice of carotenoids for dietary intake. Ascorbic acid has antioxidant activity and maintains cellular membrane activity. Ascorbic acid inhibits the conversion of cancer-causing compound N-nitroso from nitrates and nitrides present in fruit and vegetables [40]. In the current study, ascorbic acid contents in the Peelu fruit were 73.9±1.1 μg·100 g-1 (Table 4). Higher ascorbic acid content in Peelu proved its great potential for promoting health benefits. Similarly, a higher amount of ascorbic (67.9 mg·100 g−1·DW) has been reported for S. persica fruit [22].


## 3.4. Biochemical Analysis

Data regarding biochemical profiling of the Peelu fruit indicated that TSS in fruit juice were8.9±1.3°Brix (Table 4). The percent TA was 0.5±0.1 (%) with a pH value of 3.9±0.1. Additionally, the ripening index ratio was 19.7±7.3. These results showed that biochemical attributes of Peelu were at par with other ignored fruits like Averrhoa carambola, Morus alba, and Syzygium cumini [41, 42], comparable with Grewia subinaequalis, and lesser than Prunus persica and Prunus armeniaca [43] suggesting the usage of this rare fruit in different value-added products.


## 3.5. Physicochemical Analysis of Drink

Data regarding various physicochemical attributes (pH and acidity) showed a significant effect of treatments and storage duration on functional drinks prepared from Peelu. However, the interaction between treatments and storage duration was nonsignificant for pH and acidity of the functional drinks. Moreover, a nonsignificant effect of treatment and storage period, as well as their interaction, was recorded for TSS (Figure1).Figure 1
Impact of treatments (T0: placebo; T1: ethanol extract containing functional drink; T2: methanol extract containing functional drink) on overall acceptability of drink during storage period. Vertical bars indicate ±standard error of means. n=3, LSD (P≤0.05) for drinks.The highest concentration of TSS (1.6°Brix) was observed in functional drink with ethanol extract of Peelu as compared to placebo and functional drink with methanol extract of Peelu (1.58°Brix and 1.58°Brix). As far as the storage period was concerned, an increasing trend was observed for TSS for the storage period. The highest concentration of TA (1.67%) was obtained after 30 days of storage period as compared to 0 days (1.5%) (Figure2(a)). An increasing trend has been observed for acidity with prolonging storage period in all treatments. This increase in acidity (%) was more in methanol extract of Peelu when compared with other treatments. After storage of 30 days drink prepared in methanol extract showed 1.03- and 1.06-fold more TA (Figure 2(b)). Among various treatments, functional dinks prepared in methanol extract exhibited the highest (3.48) value for pH. A significant linear decreasing trend (1.13-fold) was observed for the pH of functional drinks with the increase in storage period from day-0 to day-30 (Figure 2(c)).Figure 2
Impact of treatments (T0: placebo; T1: ethanol extract containing functional drink; T2: methanol extract containing functional drink) on total soluble solids (a), acidity (b), and pH (c) of functional drink during the storage period. Vertical bars indicate ±standard error of means. n=3, LSD (P≤0.05) for drinks.Previously, the average TSS documented was about 10.4°Brix in a diet drink of apple prepared with artificial sweetener. Similarly, the results of the current investigation are in corroboration with González-Molina et al. [44], who reported a nonmomentous effect of storage period on TSS of apple and pomegranate juices and their blending. The result of the present study for pH and acidity of functional drinks is similar to Ahmed [45], who observed that a negative correlation exists between acidity and pH of juice during storage. Moreover, these increase and decrease in acidity and pH are closely linked with each other as reported by Klimczak et al. [46]. Similarly, an increase in acidity and a sharp decline in pH was monitored in fruit drink during 90 days of storage [47]. The inverse relationship between pH and acidity has also been observed in yogurt-like beverages including cereals [48]. This increase in acidity may be due to the breakdown of artificial sweeteners as well as citric acid in replacement of sugar for making therapeutic drinks [49]. Additionally, a decrease in pH and increase in acidity may be due to the degradation of polysaccharides and oxidation of reducing sugars that results in the production of acidic components. Furthermore, the development of uronic acid due to the breakdown of pectin may also be the reason for a change in the abovementioned parameters [50].


## 3.6. Sensory Evaluation

Treatments, storage period, and interaction were nonsignificant for sensory parameters of functional drinks. Color of the product/physical appearance is considered a prime factor for selection/rejection of the product by the consumer. With the increase in storage duration color score of the functional drinks was significantly reduced. The color score of the drinks was more at 0-day (6.6), while rescued to 6.1 after 30 days of storage. Among the treatment, the highest color score was obtained fromT2 (6.51±0.07) as compared to T1 and T0 (6.45 and 6.23, respectively) (Figure 3(a)). The flavor of drinks was found higher (7.1±0.06) at 0-day, while the lowest value (6.96) was recorded after 30 days of storage (Figure 3(b)). As far as various treatments are concerned, drink prepared with ethanol extract (T1) showed a better flavor score (7.14), as compared to drinks prepared with methanol extract (7.09) and drink without extract (6.94). Average values documented for sweetness and sourness at 0-day were 7.20 and 7.68, 7.60 and 7.79, and 7.36 and 7.5, respectively, whereas mean values for sweetness and sourness after 30 days of storage were 7.08 and 7.38, 7.22 and 7.5, and 7.18 and 7.39 for T0, T1, and T2 treated drink accordingly. The overall acceptability means a score of functional drinks was recorded about 7.77 during storage.Figure 3
Impact of treatments (T0: placebo; T1: ethanol extract containing functional drink; T2: methanol extract containing functional drink) on color (a), flavor (b), sweetness (c), and sourness (d) of functional drink during storage period. Vertical bars indicate ±standard error of means. n=3, LSD (P≤0.05) for drinks.Our results were in line with Ahmed [45] who reported that sensory evaluation of functional drinks prepared with polyphenol extracts declined in various sensory attributes including color, flavor, sweetness, sourness, and overall acceptability. This may be due to the reason that polyphenols induce color variation because of color imparting bodies linked with the plant polyphenols [51].


## 4. Conclusion

The Peelu fruit is a good source of macro- and micronutrients and has a sufficient quantity of biochemical contents (SSC, TA, SSC : TA ratio, and pH). Like other fruits, the juice of the Peelu fruit is a rich source of phenolic and antioxidants, including ascorbic acid and caroteniods. However, the sensory attributes of the functional drinks were in corroboration with control representing its potential for furtherin vivo study. Conclusively, Peelu exhibit high nutritional potential and could be included in the food chain with value addition.



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*Source: 5707953-2022-10-14.xml*