Preliminary Phytochemical Screening and Infra-Radiation Test of Crude the Leaves of Nothophanax scutellarium Merr., Moringa oleifera and Piper betle L.

 

Cut Sriyanti^{1, 2}, Tongku Nizwan Siregar³*, Mudatsir⁴, Azhari Gani⁵

¹Graduate School of Mathematics and Applied Sciences, Universitas Syiah Kuala, 23111,

Banda Aceh, Indonesia.

²Poltekkes Kemenkes Aceh, Darul Imarah, 23213, Aceh Besar, Aceh, Indonesia.

³Laboratory of Reproduction, Faculty of Veterinary Medicine, Universitas Syiah Kuala, 23111,

Banda Aceh, Indonesia.

⁴Department of Microbiology, Faculty of Medicine, Universitas Syiah Kuala, 23111, Banda Aceh, Indonesia.

⁵Department of Internal Medicine, Faculty of Medicine, Universitas Syiah Kuala, 23111, Banda Aceh, Indonesia

 

ABSTRACT:

Biodiversity in Indonesia is home to medicinal plants in the world, including Nothopanax scutellarium Merr., Moringa oliefera Lam., Piper betle L. Several researchers have examined the active compounds contained in these plants. This study focused on preliminary phytochemistry and FTIR analysis of leaf extracts of Nothopanax scutellarium Merr., Moringa oliefera Lam., Piper betle L. Phytochemicals were extracted with ethanol 70% by maceration. The results showed that the active compounds of flavonoids, steroids, tannins, phenols, saponins, and alkaloids were present in the three leaves. FTIR analysis observed the main peak at wavenumber 3623.44 cm^-1 in the mangkokan leaf extract, wavenumber 3690.64 cm^-1 in the moringa leaf extract, and wavenumber 3683.89 cm^-1 in the piper betle leaf extract. All of which showed the O-H phenol functional group.

 

 

 

INTRODUCTION:

Indonesia is one of the countries bestowed with biodiversity and is home to about 80% of medicinal plants globally. This rich diversity is because Indonesia is an archipelago and tropical region¹. As for some medicinal plants, namely mangkokan (Nothopanax scutellarium merr.), kelor (Moringa oliefera) and sirih (Piper betle), which usually grows in the yard. In daily life, Indonesian people also use mangkokan and moringa leaves for cooking as vegetables, while betel leaf is often used for betel nut and overcoming health problems.

 

Mangkokan belongs to the Araliaceae tribe, a shrub, grows upright, 1-1.5 m high, has woody stems, branches, small, round, long, straight. The leaves are single, stemmed, relatively thick, shaped like a bowl. Dark green leaf edges serrated and heart-shaped base. In Thailand, it is known as patios, saucer leaf, shell leaf, while in Indonesia, it is called mangkokan^2,3. In mangkokan plants commonly used as medicine are leaves, usually to treat mastitis, skin wounds, difficulty urinating, hair loss^2,3,4, fever, headaches⁵, and constipation⁶. Mangkokan leaves have alkaloid compounds, coumarin, saponins, flavonoids, and tannins in their content^7,8,9.

 

The magic tree is the name for moringa oleifera¹⁰, a family of Moringaceae^{11, 12}. Moringa is a small or medium-sized tree that proliferates, about 10-12 m high, has soft and brittle trunks, leaves as big as fingertips that are ovoid and arranged together. This plant blooms all year round and is white. The fruit contains a triangle with a 30 cm and whitish bark^10,12,13. In England, it is known as moringa, drumstick, ben-oil, tree, clarifier tree, Vietnam (chum nay), ma-rum (Thailand), malunggay (Philippines), and Indonesia is known as kelor. Moringa is a medicinal plant widely used for traditional medicine^11,14. Phytochemical screening showed that Moringa leaves contain alkaloid compounds, flavonoids, anthraquinones, tannins and phenols, glycosides^10,15,16. Various parts of the moringa plant are used to treat psychosis, eye diseases, fever, and an aphrodisiac¹⁰, wounds⁵. Traditionally the roots are applied as a plaster to reduce swelling and rheumatism. Roots, flowers, fruit, and leaves have analgesic and anti-inflammatory activity¹⁰.

 

Betel is cultivated in Sri Lanka, Indonesia, India, Malaysia, Philippine Islands, and East Africa. This plant belongs to the Piperaceae family¹⁷. Betel vine grows, up to 15 m high, depending on the medium for propagating. The stem is greenish-brown, rounded, and wrinkled, which is where the roots come out. The leaves are heart-shaped, tapered, grow alternately, stemmed, have a slightly rough texture when touched, and emit a pleasant (aromatic) smell when squeezed. Leaves are 6-17.5 cm in long and 3.5-10 cm wide, green to dark green. The roots of the riding are round in yellowish-brown¹⁸. The Arabs called betel with the names tanul or tanbul, Chinese (ju jiang, tu bi ta, tu wei teng), paan (India), betle, betle pepper, betle-vine (English), bakik serasa (Malaysia), and naagavallii (Nepal), Nagballi, Tamalapaku (Telugu), Ventrilai (Tamil), Nagarbael (Gujarati) and sirih (Indonesia)^17,18,19,20. Betel leaf contains alkaloid compounds, flavonoids, steroids, saponins, phenols, eugenol, carbohydrate, and amino acids^20,21,22. The leaves have been used in traditional medicine, among others for filariasis, obesity, coughing, tightness, indigestion, the release of breast milk, inflammatory swelling (orchitis, arthritis, and mastitis), bad breath, body odor, preventing tooth decay, relieving vaginal itching, stopping bleeding in the nose, curing eczema, lymphangitis, asthma, rheumatism, wounds, stomach medication²⁰, fever and headache⁵.

 

This study focuses on preliminary phytochemicals and FTIR tests of mangkokan, moringa, and betel leaves. This study is expected to provide evidence to support the use of traditional medicinal plants. The results can help clarify the comparison of the three leaves' active compound content and functional groups.

 

MATERIAL AND METHODS:

Studi Area:

The plant samples used were mangkokan leaves collected from Gampong Neusu Aceh, Baiturrahman, Banda Aceh with coordinates 5^o32’10” N and 95^o19’3” E. Moringa leaves were collected from Gampong Lampeuneurut Ujong Blang, Darul Imarah, Aceh Besar at coordinates 5^o30’43” N and 95^o19’38” E. Betel leaves were taken from Gampong Lambhuk, Ulee Kareng, Banda Aceh at coordinates 5^o33’4.72” N and 95^o20’10.04” E. We make sure the plants are healthy and not infected. The identification of herbarium samples was carried out at the Biology Laboratory of the Faculty of Mathematics and Natural Sciences, Syiah Kuala University, Banda Aceh.

 

Preparation of Leaf Extracts:

Samples of mangkokan leaves, moringa leaves, and fresh betel leaves were collected in April 2020, from each location of approximately 5 kg, then washed using running water to remove dirt and foreign particles, then air-dried and ground into powder. Each sample was weighed 300 g for maceration with 70% ethanol for seven days. Then filtered to obtain macerate, evaporated using a rotary evaporator until a thick extract was obtained, then phytochemical analysis and FT-IR Spectroscopic were performed.

 

Phytochemical Analysis:

All extracts were screened for preliminary phytochemicals following the procedure^{23, 24, 25, 26, 27, 28, 29} to test for the presence of flavonoids, steroids, terpenoids, tannins, phenols, saponins, and alkaloids.

 

Identification of Flavonoid compounds :

Three milliliters of the extract is added with Mg powder, then add 3-4 drops of concentrated HCl. The formation of red or orange color indicates the presence of flavonoids.

 

Identification of Steroid compounds :

Two milliliters of the extract solution was added ten milliliters of chloroform, then added one milliliter of anhydrous acetic acid and 2-3 drops of concentrated sulfuric acid. Green discoloration indicates the presence of steroids.

 

Identification of Terpenoid compounds :

The identification for terpenoids is the same as for the steroid test. The appearance of red or purple color indicates the presence of a terpenoid.

 

Identification of Tannin compounds :

1% gelatin solution was added to the extract. The formation of a white precipitate indicates the presence of tannins.

 

Identification of Phenol compounds :

One milliliter of the extract was dissolved with two milliliters of 5% FeCl₃ solution. The formation of a bluish-black color indicates the presence of phenol.

 

Identification of Saponin compounds :

One milliliter of the extract is added to nine milliliters of distilled water then shake vigorously for 15 seconds. The presence of saponins is indicated by the persistence of the resulting foam for ten minutes.

 

Identification of Alkaloid compounds :

The three-milliliter extract solution was added with one milliliter of 2N HCl, then separated into four test tubes. Tube 1 as a comparison, tube 2 was added with dragendorff reagent, tube 3 was added with major reagent, and tube 4 was added with Wagner's reagent. Positive results if there is a red brick deposit with dragendorff reagent, beige or greenish with Mayer reagent, and brown with Wagner reagent.

 

FTIR Test :

Fourier Transform Infrared Spectroscopic (FTIR) is a tool for identifying the types of chemical bonds or functional groups present in phytochemistry. The light absorption wavelength is a characteristic that arises from the chemical bonds seen in the annotated spectrum. By interpreting infrared absorption, the chemical bonds in a compound can be determined^{30, 31, 32, 33, 34, 35, 36}.

 

Table 1: The results of initial phytochemical screening of 70% ethanol extract from leaves of Nothophanax scutellarium merr., Moringa oleifera, and Piper betle linn.

 

 

RESULTS AND DISCUSSION :

Preliminary Phychemical Analysis :

The results of the preliminary phytochemical screening showed that the 70% ethanol extract of the leaves of Nothophanax scutellarium merr., Moringa oleifera and Piper betle linn. contained flavonoids, steroids, tannins, phenol, saponins, and alkaloids, as can be seen in Table 1.

 

FTIR Analysis :

FTIR spectrum of Nothophanax scutellarium merr. leaf extract :

FTIR spectrum data of Nothophanax scutellarium merr. leaf extract shows phenol content, which is strengthened by the OH group's presence on the sharp band absorption shown in the wavenumber 3623.44 cm^-1 and the sharp band absorption with the wave number 3754.60 cm^-1. At wave number 2958.93 cm^-1 and the shape of the acute band, it shows an alkane group in the form of C-H, while at wave number 2378.33 cm^-1 with a sharp band absorption indicates the presence of a C=O ester compound. There is also a C=C alkene group with a sharp band absorption at wavenumbers 1700.32 cm^-1 and 1912.50 cm^-1. There is an aromatic group, namely nitro aromatic C-NO₂, shown in the absorption band that is sharp with a wavenumber 1457.89 cm cm^-1 and 1424.49 cm^-1. The C-O-C ether group shows the wavenumber at 1121.65 cm^-1, indicated by the sharp band absorption. At wave number 852.57 cm^-1 indicates the presence of an aliphatic (CH₂)n group, as shown in Fig. 1 and Table 2.

 

FTIR spectrum of Moringa oleifera leaf extract :

Based on FTIR spectrum data of Moringa oleifera leaf extract, there is an uptake in the area of ​​3690.64 cm^-1 and 3768.46 cm^-1 with a sharp band shape, indicating the presence of phenol content in the form of OH. At wave number 2966.68 cm^-1 with the shape of the sharp band, it shows an alkane group in the form of C-H, while at wave number 2369.04 cm^-1 with a sharp band absorption indicates the presence of a compound C=O ester.

 

Fig. 1: FTIR analysis of Nothopanax scutellarium merr. leaf extract

Table 2: The results of FTIR Nothophanax scutellarium merr. leaf Extract

 

Then, C=C alkene group with a sharp band absorption at wave number 1693.57 cm^-1. There is an aromatic group, namely nitro aromatic C-NO₂, which is shown in the sharp band absorption with a wavenumber of 1436.07 cm^-1 and 1518.04 cm^-1. The ether group C-O-C shows the wavenumber at 1135.16 cm^-1, which is indicated by sharp band absorption, while at the wavenumbers 1248.96 cm^-1 and 1294.29 cm^-1 with a sharp band shape. Furthermore, the wavenumbers 662.27 cm^-1, 747.45 cm^-1, 801.46 cm^-1, and 849.68 cm^-1 with a sharp band indicate the aliphatic presence (CH²)n groups, as shown in Fig. 2 and Table 3.

 

 

 

FTIR spectrum of Piper betle linn. leaf extract :

From the FTIR spectrum data of Piper betle linn. leaf extract, there is an uptake in 3683.89 cm^-1 and 3683.89 cm^-1 with a sharp band shape, indicating phenol content in the form of OH. At wave number 2960.86 cm^-1 with a sharp band, it indicates an alkane group in the form of C-H. At wave number 2374.47 cm^-1 with a sharp band absorption indicates a C=O ester compound, while at wave number 1693.57 cm^-1 with a sharp band absorption indicates the presence of C = C alkene groups. There is also an aromatic group, namely nitro aromatic C-NO₂, shown in the sharp absorption band with a wavenumber of 1452.46 cm^-1. The C-O-C ether group shows the wavenumber at 1126.48 cm^-1, which is indicated by a sharp band absorption.

 

 Fig. 2: FTIR analysis of Moringa oleifera leaf extract                               

 Fig. 3: FTIR analysis of Piper betle linn. leaf extract

 

Table 3: The results of FTIR Moringa oleifera leaf Extract

Table 4: The results of FTIR Piper betle linn. leaf Extract

 

In addition, the wavenumbers 561.31 cm^-1, 743.59 cm^-1, and 806.28 cm^-1 with the sharp band shape indicated the presence of aliphatic (CH2) n groups. As shown in Fig. 3 and Table 4.

 

DISCUSSION :

Preliminary phytochemical tests were carried out on the ethanol extract of Nothophanax scutellarium merr. leaves, and it is found the presence of flavonoid compounds, steroids, tannins, phenols, saponins, and alkaloids. These findings follow⁹, but Eden reported no alkaloids and phenols in methanol extract, chloroform fraction, n-hexane, and water fraction⁷. However, they agree with flavonoid and saponin extracts. Herawati also reported the absence of phenols in the leaf extract of Nothophanax scutellarium merr⁸. Then in the initial phytochemical test on the ethanol extract of Moringa oleifera leaves, it was found that there were flavonoids, steroids, tannins, phenols, saponins, and alkaloids. These results are in line with^37,38,39, while Abubakar and Usman reported no steroid content in methanol and water extracts¹⁵. However, they agree with the alkaloids, tannins, flavonoids, saponins, and phenols in Moringa oleifera leaf extract. Furthermore, secondary metabolites were examined with the results that there were flavonoids, steroids, tannins, phenols, saponins, and alkaloids in the ethanol extract of Piper betle linn. leaves. This study is following^40,41,42, while according to the reports of Avijit and Kaveti no flavonoids were found in Piper betle linn. leaf extract^19,43.

 

The chemical content of each plant species varies according to ecology, soil, and climate⁴⁴ and the maturity at which leaf samples are collected⁴⁵. It should be noted that the phytochemical components are important factors that determine the antimicrobial properties of leaf extracts. It explains why medicinal plants are used as antimicrobial drugs. Some authors have linked bioactive compounds with the antimicrobial properties of plant extracts⁴⁶.

 

FTIR spectroscopy studies revealed that the ethanol extract of mangkokan leaves showed the presence of functional groups OH (phenol), CH (alkane), C=O (ester), C=C (alkene), C-NO₂ (aromatic nitro), COC (ether) and (CH₂)n (aliphatic), while research conducted by Syafrinal⁴⁷ using n-hexane, ethyl acetate and methanol solvents reported the functional groups O-H, C-H, and C=C. The FTIR results of the ethanol extract of Moringa leaves confirmed the functional groups O-H, C-H, C = O, C = C, C-NO₂, C-O-C, and (CH₂)n. This result is in line with the research of Cruz-Espinoza⁴⁸, except for C=O, C-NO₂, C-O-C, and C-H. The functional groups in betel leaf extract were O-H, C-H, C=O, C=C, C-NO₂, C-O-C, and (CH₂)n. This study is in line with⁴³, except for C-O-C, CH₃, and C-NO₂. The functional groups in the extracts of mangkokan, Moringa, and betel leaves are based on the absorption bands obtained at specific wavenumbers by FTIR spectroscopic analysis. Several differences were observed in the extracted spectrum of the three extracts.

 

CONCLUSION :

From this study, it is concluded that there is a correlation with previous reports on phytochemical compounds in the ethanol extract of the leaves of Nothophanax scutellarium merr., Moringa oleifera and Piper betle Linn. When the extract was passed to FTIR, the functional groups of the components were separated based on their peak ratio, as for the FTIR analysis results of the ethanol extract of the leaves of Nothophanax scutellarium Merr., Moringa oleifera and Piper betle L. confirm the presence of functional groups O-H, C-H, C = O, C = C, C-NO₂, C-O-C, and (CH₂)n. Thus in this study, a valuable active component can be used for various therapeutic purposes.

 

ACKNOWLEDGEMENT:

All authors would like to thank all parties for their assistance in this research.

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

 

 

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Accepted on 29.08.2021               © RJPT All right reserved

Research J. Pharm.and Tech 2022; 15(4):1805-1810.