A Comprehensive Review of Phytochemical Profile and Pharmacological Attributes of Epiphyllum Oxypetalum

The plant kingdom includes a wide variety of species, producing a diversity of bioactive molecules with different chemical scaffolds. Throughout the centuries, medicinal and aromatic plants have remained a significant part of everyday life, even with the advancements in modern medicine and the pharmaceutical industry. Moreover, plants serve as a rich and invaluable source of natural remedies, many of which have long been utilized in traditional healing systems. However, there are numerous plant species with unexplored medicinal potential, and many others yet to be discovered. Epiphyllum Oxypetalum is a species belongs to the family Cactaceae. Epiphyllum Oxypetalum, originally found in Central America and Mexico, has a long-standing history of being used for its medicinal properties. It is one of the native species with a long history of useful therapeutic properties. Brahma Kamal, Nishagandhi, and night blooming cereus are some of the common names for Epiphyllum Oxypetalum [2]. The Epiphyllum Oxypetalum plant can reach a height of 2 to 6 meters and features dark green foliage. The woody, densely branched trunks and stems of these plants can reach a diameter of 2 cm or more. The leaves of these plants are lancet-shaped and low sideways. Glossy green leaves on the underside and upper surface of sharply pointed leaves, thinning, wavy and serrated leaves, narrow leaves arranged in a line with 1.6 to 1.8 mm of interest on top, nocturnal (blooming at night), funnel-shaped and fragrant [3]. Epiphyllum Oxypetalum is primarily grown for ornamental purposes, but it is now also being investigated for its antibacterial and nutritional benefits [4]. This medicinal plant includes a wide range of chemicals with antioxidant and therapeutic qualities that could serve as a basis for the creation of new medications. The majority of medicinal plants contain substances called alkaloids, tannins saponins, terpenoids, flavonoids, and phenolics that are thought to have developed naturally in the plant as a means of defence against various pests and diseases [5].

Figure 1: Plant in flowering stage and plant with leaves showing structure of   Epipyllum Oxypetalum

Numerous researchers have studied Epipyllum Oxypetalum phytochemically, and several active chemical components have been identified upon isolation. Chemical components such as Alkaloids, saponins, terpenoids, protein, amino acids, steroids, flavonoids, tannins ^[6] and triterpenoids ^[7] were present in the flowers. Carbohydrates, phenols, sterols, glycosides, resins ^[8] were present in the leaves.

Table 4: Chemical constituents of Epiphyllum Oxypetalum

Phytochemical Analysis of flower extract:  

Qualitative chemical analysis was conducted on all extracts of Epiphyllum Oxypetalum flowers to identify the presence of various phytoconstituents, as detailed below:

1ml of the extract was stirred with 1ml of dil. Hcl and filtered. The filtrate was tested with various reagents as follows ^[9].

Qualitative chemical analysis was conducted on all extracts of Epiphyllum Oxypetalum leaves to identify the presence of various phytoconstituents, as detailed below ^[10]

Epiphyllum Oxypetalum plant species has shown various activities as that in picture:

Figure 2: Pharmacological activity of Epiphyllum Oxypetalum

Infectious microorganisms like bacteria, viruses, or fungi typically cause inflammation when they enter the body, settle in certain tissues, or move through the bloodstream. Additionally, events including tissue damage, cell death, malignancy, ischaemia, and degeneration can all cause inflammation. During various forms of inflammatory responses, a large number of inflammatory mediators are produced and released. Pro and anti-inflammatory mediators are the two primary categories into which inflammatory chemicals are typically separated ^[11].  The fresh plant of Epiphyllum Oxypetalum was collected and authenticated, then the fresh leaves were shed dried and coarsely powdered in a mechanical grinder. Successive, extraction was carried out using ethanol and water in Soxhlet apparatus. After complete solvent removal, the extract weighed and stored in a desiccator until further use. The Anti-inflammatory activity of Epiphyllum Oxypetalum ethanolic extract was evaluated on rats. Both sexes of adult Albino Wistar rats weighing 150–200g were chosen. The rats were given a week to get used to the lab environment before the treatment. The animals were kept in polypropylene cages with a maximum of four animals per cage under conventional laboratory settings, which included a temperature of 25ºC ± 2ºC, a relative humidity of 50±5%, and a photoperiod of 10 hours dark and 14 hours light. The use of animals in the experiments was approved by the institutional animal ethics committee. The department of animal ethics committee accepted the study's protocol, and the procedures were conducted in accordance with CPCSEA criteria.

Experimental design:

The study was carried out by using six groups of six animals

percentage of paw oedema inhibition = (Vt-Vo) control - (Vt-Vo) treated X 100

(Vt-Vo) control

where Vt is the rat paw volume at time "t"

• Vo is the rat paw volume at the beginning (prior to carrageenan injection)
• (Vt-Vo) control is the oedema generated in the control group, and
• (Vt-Vo) treated is the oedema generated during treatment.

In-vitro evaluation of anti-inflammatory activity:

1. Membrane stabilisation method

• Preparation of human Red Blood Cells (HRBC) suspension 
• Heat induced haemolytic activity

2)  Inhibition of albumin denaturation

Anti-Oxidant Activity:

The radical scavengers which protect the human body against free radical that causes pathological condition is known as anti-oxidants. The present concept of toxicity is due to the participation of oxygen free radicals or reactive oxygen species. Therefore, it becomes essential to test the antioxidant potential of the plant material [13]. Natural anti-oxidants or phytochemical anti-oxidants are the secondary metabolites of plants [14]. Some of the antioxidants produced by the plants include carotenoids, flavonoids, tocopherols, Beta carotene, Lycopene, Sesamol, Anthocyanins, Catechins, Ellagic acid, Lutein, Resveratrol, cinnamic acids, benzoic acids, folic acid, ascorbic acid, tocotrienols etc.,[15]. The Epiphyllum Oxypetalum belongs to the cactaceae family have much attention towards the development of the ethno medicines with strong anti-oxidant properties. The fresh plant of Epiphyllum Oxypetalum was collected and authenticated, then the fresh leaves were shed dried and coarsely powdered in a mechanical grinder. Successive, extraction was carried out using ethanol in Soxhlet apparatus at 40-50?. the extraction process is carried out at 80?.

The Anti-inflammatory activity of Epiphyllum Oxypetalum ethanolic extract was evaluated by using two methods:

1) DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging assay method

2) Hydrogen peroxide scavenging assay method

1. DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging assay method:

A stable radical called DPPH was used to test the hydrogen-donating or radical-scavenging capabilities of alcohol and aqueous extracts as well as the standard vitamin C, L-ascorbic acid. A 0.1 mM DPPH solution in alcohol was made, shielded from light by keeping it dark, and folded with aluminium foil. 3ml of this solution were then added to 1ml of extracts at different concentrations (100–2000 μg/ml) or a standard solution of 10–100 μg/ml. After 30 minutes, absorbance was measured at 517 nm.

The percentage inhibition activity was calculated using the following formula:

% Inhibition = [(A control – A test)/A control] ×100

Were,

• A control- Absorbance of the control 
• A test - Absorbance of extract/standard taken as ascorbic acid

Assay: 

Table 9: DPPH scavenging activity of standard drug ascorbic acid

Figure 3: Graphical representation of DPPH radical scavenging assay of alcohol and aqueous drug

Figure 4: Graphical representation of DPPH radical scavenging assay of standard drug ascorbic acid

2. Hydrogen peroxide scavenging assay method:   

This assay method was used to determine the extract's capacity to scavenge hydrogen peroxide. 4ml of extract was prepared using distilled water at different concentrations (100 μg to 500 μg). The extracts were then combined with 0.6 millilitres of a 4 mM H2O2 solution made in phosphate buffer (0.1 M with pH 7.4) and incubated for ten minutes. The absorbance was measured at 230 nm against a blank solution that contained the plant extract without H2O2. Ascorbic acid was used as a standard reference.

The percentage inhibition activity was calculated using the following formula –

% Inhibition = [(A control – A test)/A control] ×100

Were, A control- Absorbance of the control 

A test - Absorbance of extract/standard taken as ascorbic acid

Assay: 

Table 11: Hydrogen Peroxide scavenging assay of standard drug ascorbic acid

Figure 5: Graphical representation of hydrogen peroxide scavenging assay of alcohol and aqueous extract of Epiphyllum Oxypetalum leaves

Figure 6: Graphical representation of hydrogen peroxide scavenging assay of standard drug ascorbic acid

ANTI-UROLITHIATIC ACTIVITY:

Urolithiasis is derived from the Greek words ouron (urine) and lithos (stone). Urolithiasis refers to the accretion of hard, solid and non-metallic minerals in the urinary tract. A hard, crystalline mineral substance that forms in the kidney or urinary tract is called a kidney stone. Renal calculi are another name for kidney stones. Nephrolithiasis is the name for kidney stone disease ^[16].   Around 12% of the world population is affected by the urolithiasis. The physiochemical process of calcified kidney stone production involves crystal nucleation, aggregation, and growth, and it is facilitated by a number of biological events, including increased calcium or sodium oxalate, urates, urine volume, and pH. According to their size, type, and location, 90% of patients with upper urinary tract stones are currently treated, with success rates ranging from 68 to 86%. Approximately 75% of all renal stones are caused by calcium phosphate or calcium oxalate, and eating more protein has been associated with an increased risk of developing renal stones ^[17].  The Epiphyllum Oxypetalum leaves was collected and authenticated. The leaves were cleaned with water, dried and grind into a fine powder using blender. Extracted with ethanol using Soxhlet extractor for 6 to 8 hours. Filter the extract using a muslin cloth, transfer it to 50 ml tubes and centrifuge it for 15 minutes at 25 °C at 4,000 rpm.

Investigation of anti-urolithiatic activity test by in-vitro using a titration method:

In the laboratory, calcium oxalate kidney stones were produced by mixing an equimolar solution of sodium oxalate in 10 ml of 2N H₂SO₄ with dehydration of calcium chloride in distilled water. Both processes were given enough distilled water to react in a beaker, producing calcium oxalate precipitate. An ammonia solution is used to remove any last traces of sulfuric acid from the precipitate, which is then rinsed with distilled water and dried at 60°C.  1gm of calcium oxalate and 10 gm of extract were combined in an egg's semi-permeable membrane to measure the calcium oxalate dissolving percentage. This was suspended with 100 cc of 0.1M Tris buffer in a conical flask. The first group only had 1 mg of calcium oxalate, serving as a control. As a positive check, the second group had 10 mg of Cystone and 1 mg of calcium oxalate.  Both the third and fourth groups have one milligram of calcium oxalate and ethanol extracts. All group's conical flasks were heated to 37°C for two hours in incubator.  The contents of the semi-permeable membranes for each group should be placed in different test tubes. 2 ml of 1N sulfuric acid is added to each test tube with acid after titrating with 0.9494 N KMnO₄ until the end point is achieved and the colour turns deep pink.  To determine the total amount of various solvent extracts of dissolved calcium oxalate, subtract the amount of calcium oxalate that was still present after the experiment's first run.

Table 12: % calcium oxalate (CaOx) dissolution by Epiphyllum Oxypetalum leaves extracts

This study evaluates the effectiveness of ethanolic extract of Epiphyllum Oxypetalum on anti-urolithiatic activity. The maximum rate of calcium oxalate "CaOx" dissolution, 80% was seen in the ethanolic extract.  It was shown that ethanolic extracts of Epiphyllum Oxypetalum were more effective at dissolving calcium oxalate.

Figure 7: Urolithiasis percentage of ethanolic extract of Epiphyllum Oxypetalum according to their concentration

ANTHELMINTIC ACTIVITY:

Helminths is a Greek word meaning “worm”. Helminth infection is common parasitic infection affecting large number of populations, especially children. Helminths could be classified into three classes as Nematodea, Cestoidea and Trematodea. A wide range of worms are observed varying size from less than 1mm -1m. People living in Australia, south-east Asia, India, Mexico, Sri Lanka, and Thailand are most affected by this infection. Helminths mainly enter through the contaminated drinking water or raw meats from infected animals. They may also enter into the body through skin by insect bites or walking and swimming in the contaminated soil and water. helminths require a living host for survival, reproduction causing physical, nutritional, cognitive impairment in young children. Anthelmintic drugs are drugs which are used for treatment of helminthic infections caused by various worms. These are the drugs which act locally to emit the worms from GIT or may also act systematically to eliminate adult helminths and prevent tissue and organs from developmental forms ^[19]. The Epiphyllum Oxypetalum leaves was collected and authenticated. The leaves were cleaned with the fresh water and dried in the shade. The dried leaves were grinded into fine powder using blender. The powder was extracted with the ethanol using Soxhlet extraction process. After extraction the extract if filtered using muslin cloth and transferred it to 50ml tubes and centrifuged for 15mins at 4000rpm at 25 ^oC. Finally, the extract is analysed for photochemical constituents.

Assessment of In-vitro Anthelmintic Activity of Epiphyllum Oxypetalum:

Pheritima Posthuma, an adult Indian earthworm, was treated with anthelmintics. The standard medication albendazole was diluted with sterile saline and then added to Petri plates concentrations of 25, 50, and 100 milligrams per millilitre. Ethanolic extracts from leaves were mixed with ordinary saline to create concentrations of 25, 50, and 100 mg/ml. The saline solution served as the negative control. Consequently, the Petri plates were filled with each of these dilutions. For the experiment, earthworms were split up into six groups (n = 6). In every Petri dish, earthworms of roughly the same size were kept at the same temperature. The point at which there was no movement of any kind other than when the worms were severely disoriented is used to calculate the duration of impairment. Once it was determined that the worms did not move when severely shaken or when submerged in warm water at 50°C, the moment of death for each individual worm was recorded. Minutes were used to express both the paralysis and death times.