**3.5. Qualitative phytochemical analysis of** *Opuntia elatior* **fruit extract**

The phytochemical constituents present in the methanolic extract of *Opuntia elatior* are phytosterols and alkaloid compounds. The presence of yellow color precipitates of test sample by Mayer's reagent shows the presence of alkaloids. The presence of golden yellow color of test sample by adding conc. H<sup>2</sup> SO4 shows the presence of phytosterols. Flavanoids, saponin, phenol and tannin are absent in methanolic extract of *Opuntia elatior*. Phytochemical constituents of the methanolic extract of *Opuntia elatior* were qualitatively tested for their presence as depicted in **Table 2**.

The presence of these phytochemicals in the plants extract enhances their pharmaceutical and therapeutic potentials. Alkaloid is reported to carry antimicrobial activities [22]. The presence of alkaloids in the investigated plants of the wild cucurbits indicates that they have medicinal values. Alkaloids have a powerful effect on the physiology of animals [23]. Sterols in modern clinical studies have shown that they play an important role as anti-inflammatory and analgesic agents [24].

**Figure 4.** Antioxidant activity of different extract of *Opuntia elatior* fruit.

extracts using DPPH method are shown in **Figure 4**. DPPH radical scavenging activity of methanolic extract of *Cantaloupe* (muskmelon) pulp shows 48.55% [18] (Ibrahim *et al.,* 2016), and *Opuntia shows* 57.37% activity. This shows that *Opuntia* has a good antioxidant potential. Itankar *et al*. reported that *Opuntia elatior* fruit has 64.14% of activity which is close to methanolic extract of *Opuntia elatior* [21]*. Opuntia* is a good source of vitamin C this helps increase antioxidant properties and reduces the risk of diseases such as atherosclerosis and cancer. Vitamin C is an electron donor. As an electron donor, it helps stabilize unpaired electrons in

**Nutrients Methanolic extract Hexanoic extract Distilled water extract**

Protein (%) 0.60 0.42 0.40 Carbohydrate (%) 1.02 0.66 0.91 Fat (%) 0.11 0.08 0.05 Vitamin C (mg/100 g) 63.29 31.64 45.21

Inflammation in the body acts as a defense mechanism. When a foreign substance enters in the body, it causes infection and injury. To protect our body from this, the infected area swells. The purpose of inflammation is to localize and eliminate the foreign substances so that the body can heal itself. Inflammation prevents excess blood flow to rich the site of damage. The anti-inflammatory activity of *Opuntia elatior* was studied using inhibition of protein denaturation method. Protein denaturation means the loss of biological properties of protein molecules. Denaturation of proteins is responsible for the cause of inflammation and its conditions like rheumatoid, arthritis, diabetes, cancer, and so on. Hence, prevention of protein

the body and reduces oxidative stress.

**Figure 3.** Extract yielded (%) of *Opuntia elatior* fruit.

132 Phytochemicals - Source of Antioxidants and Role in Disease Prevention

**Table 1.** Nutritional value of *Opuntia elatior* fruit extract.

**3.4. Anti-inflammatory properties of** *Opuntia elatior* **extract**

denaturation may also help in preventing inflammatory conditions.

**Figure 5.** Inhibition of protein denaturation, of different extract of *Opuntia elatior* fruit.


**Table 2.** Phytochemical constituents of *Opuntia elatior* fruit extract.

#### **3.6. Thin layer chromatography of** *Opuntia elatior* **fruit extract**

Methanolic extract obtained from *Opuntia elatior* fruit was carried out to thin layer chromatography to establish the purity and composition of materials. The component present in methanolic fruit extract was identified by the concentrated H2 SO4 test. After separation of methanolic fruit extract, one band was observed. The band observed under UV light is shown in figure–3.8. R*f* value of the band was measured that is 0.85. After application of H2 SO4 band, H2 SO4 was applied to the band to check the presence of compound. Brownish color of the band was observed after applying of H<sup>2</sup> SO4 . This brownish color of the band shows the presence of alkaloid in the methanolic extract of *Opuntia elatior.*

> gives quantitative assessment of a compound. Peak size of the area is nearly equal to the amount of the substance present in the mixture. For the identification of the chemical profile, the sample was subjected to further characterization and isolation of compound. In this study,

> GC-MS analysis of methanolic extract of *Opuntia elatior* is shown in **Figure 7**. The separation techniques coupled with GC-MS allowed separation of constituents as shown in the GCMS

total nine bands were obtained in the sample.

**Figure 7.** GC-MS chromatogram of methanolic extracts of *Opuntia elatior.*

**3.8. GC-MS analysis of methanolic extract of** *Opuntia elatior* **fruit**

**Figure 6.** HPTLC profile and chromatogram of methanolic extract of *Opuntia elatior* fruit.

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#### **3.7. HPTLC analysis of methanolic extract of** *Opuntia elatior* **fruit**

For the HPTLC analysis the extract of *Opuntia elatior* was run in a mobile phase of methanol:chloroform (8:2). The plate was visualized at 254 nm. HPTLC profile of methanolic extract of *Opuntia elatior* shows three bands at 254 nm, it is shown in **Figure 6**. Maximum R*f* values were obtained by track 1. Different peak spectral comparison of methanolic extract is shown in **Figure 5**. This shows the different compounds present in the sample. Each compound has its own unique pattern. Determination of the peak height of chromatographic peak Evaluation of Nutritional and Medicinal Properties of *Opuntia elatior* Mill http://dx.doi.org/10.5772/intechopen.77081 135

**Figure 6.** HPTLC profile and chromatogram of methanolic extract of *Opuntia elatior* fruit.

**Figure 7.** GC-MS chromatogram of methanolic extracts of *Opuntia elatior.*

**3.6. Thin layer chromatography of** *Opuntia elatior* **fruit extract**

**Table 2.** Phytochemical constituents of *Opuntia elatior* fruit extract.

**Phytochemical constituent Color observed Methanolic extract Distilled water** 

**Figure 5.** Inhibition of protein denaturation, of different extract of *Opuntia elatior* fruit.

134 Phytochemicals - Source of Antioxidants and Role in Disease Prevention

Alkaloids Yellow color ppt + — + Flavanoids Purple to cherry red — — — Saponin Presence of foam — — — Phytosterols Golden yellow + + + Phenol Blue green — — — Tannin White ppt — — —

methanolic fruit extract was identified by the concentrated H2

ence of alkaloid in the methanolic extract of *Opuntia elatior.*

**3.7. HPTLC analysis of methanolic extract of** *Opuntia elatior* **fruit**

band was observed after applying of H<sup>2</sup>

H2 SO4

Methanolic extract obtained from *Opuntia elatior* fruit was carried out to thin layer chromatography to establish the purity and composition of materials. The component present in

methanolic fruit extract, one band was observed. The band observed under UV light is shown

For the HPTLC analysis the extract of *Opuntia elatior* was run in a mobile phase of methanol:chloroform (8:2). The plate was visualized at 254 nm. HPTLC profile of methanolic extract of *Opuntia elatior* shows three bands at 254 nm, it is shown in **Figure 6**. Maximum R*f* values were obtained by track 1. Different peak spectral comparison of methanolic extract is shown in **Figure 5**. This shows the different compounds present in the sample. Each compound has its own unique pattern. Determination of the peak height of chromatographic peak

was applied to the band to check the presence of compound. Brownish color of the

in figure–3.8. R*f* value of the band was measured that is 0.85. After application of H2

SO4

SO4

**extract**

. This brownish color of the band shows the pres-

test. After separation of

SO4

**Hexanoic extract**

band,

gives quantitative assessment of a compound. Peak size of the area is nearly equal to the amount of the substance present in the mixture. For the identification of the chemical profile, the sample was subjected to further characterization and isolation of compound. In this study, total nine bands were obtained in the sample.

#### **3.8. GC-MS analysis of methanolic extract of** *Opuntia elatior* **fruit**

GC-MS analysis of methanolic extract of *Opuntia elatior* is shown in **Figure 7**. The separation techniques coupled with GC-MS allowed separation of constituents as shown in the GCMS trace in **Figure 7**. Compounds are not separated properly because of its higher background value. Because of higher background peaks are not seen clearly. The peaks start forming at 70% and continued for minutes. The identifications of phytochemical compound were based on the peak area, retention time and molecular formula. The GC-MS data can be used to identify major bioactive, phytochemical constituent. The GC-MS analysis of the fruit sample revealed the presence of 19 compounds. Thus, the presence of unidentified compound initiated further investigation on the fruits of *Opuntia elatior*. Further analysis of the fruit extract is being done for the NMR studies for structural analysis of the compound. Thus, the presence of alkaloids and phytosterols present in the methanolic fruit extract of *Opuntia elatior* open ample scope for further investigations. Thus, chemical present of alkaloids and phytosterols present give activity of antioxidant and anti-inflammatory. The different author reported the ethanolic extracts present the 14 compounds compared to the methanolic extracts 19 chemical compound presents. This indication more support to the further analysis of the fruit extract is required for the NMR studies for structural analysis of the compound identification. It gives better idea of which chemical compound is responsible for the antioxidant and anti-inflammatory activity.

**Author details**

**References**

Krishna N. Patel and Kalpeshkumar B. Ishnava\*

Sciences, New Vallabh Vidyanagar, India

publicaccess.nih.gov

2015;**44**(10):1309-1321

Technology. 2012;**49**:530-536

365-370

Distributors, Dehradun; 1999. pp. 1173-1178

\*Address all correspondence to: ishnavakb203@yahoo.com

Ashok and Rita Patel Institute of Integrated Study and Research in Biotechnology and Allied

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