In general, species in the Leea genus prefer moist, fertile, well-drained soil and flourish in full or partial sunlight. The stems of the perennial plant Leea macrophylla might get a little woody. From a tuberous rootstock, it frequently takes on the appearance of a shrub and sometimes a tree. For use locally as food, medicine, and color, this plant is collected from the wild. The Leea genus has been shown to have therapeutic applications in the treatment of sexual impairment, bodily aches, cancer, and diarrhea [1]. Lipid profiles, total protein, and CK were used to assess the hepatoprotective effect of Leea macrophylla leaf extract on rats.-MB [2].  Anti-inflammatory activity [3] Antidiabetic activity [4] Antimicrobial activity [5], wound healing properties  [6] gasrtopotective activities [7] of leaves extract were also reported.

The current study uses GC-MS techniques and preliminary phytochemical analysis of Leea macrophylla roots to analyze phytochemical diversity. In order to estimate the plant extract’s antibacterial activity, its inhibitory action against certain microorganisms was also examined. A case study was performed to evaluate the bone healing property of roots of Leea macrophylla.

Material and Methods

Collection of Plant material

Leea macrophylla roots were collected from the Melghat forest’s Semadoh area in the Indian state of Maharashtra’s District Amravati. After being transported to the lab, root tubers were allowed to dry in the shade. After drying, the tubers are ground into a powder and stored in an airtight bottle until they are needed again.

Extract Preparation

The Soxhlet extraction method was used to carry out the phytochemical extraction. A total of six solvents were employed for extraction: petroleum ether (PE), chloroform (Cl), ethyl acetate (EA), acetone (ACE), methanol (M), and water (WA). Each solvent was then circulated through the plant material for six hours, continuing until the solvent in the upper chamber became colorless. This process typically takes about twenty-four hours, and the extracted materials were then kept in a refrigerator at forty degrees Celsius until they were needed.

Preliminary Phytochemical Tests

Leea macrophylla underwent a preliminary phytochemical investigation. Alkaloids, flavonoids, terpenoids, phenol, tannin, anthocyanin, anthraquinones, lipids, carbohydrates, amino acids, saponins, sterols, and heart glycoside were all tested for.

GCMS Analysis

The research was done using Gas Chromatography-Mass Spectrometry (GCMS) to better understand the chemical makeup of the plant extracts.The Indian Institute of Technology in Powai, Mumbai, Maharashtra, India, conducted this thorough analysis.

Observation and Results

Prilimanry phytochemical analysis – Using petroleum ether, chloroform, ethyl acetate, acetone, methanol, and water extracts, a phytochemical analysis of the root tuber of Leea macrophylla revealed the presence of carbohydrates, protein and amino acids, fats, phenols, terpenoids, alkaloids, flavonoids, anthocyanins, and anthraquinones, with the exception of cardiac glycosides.

Chloroform and methanol extract of L. macrophylla root reported presence of carbohydrates. Petroleum ether extracts diagnose proteins and amino acids in all conducted tests. Methanol and aqueous extract indicate occurrence of fats. Only aqueous extract of L. macrophylla root was rich in saponins, whereas petroleum ether and chloroform fractions showed presence of terpenoids. Except for acetone and methanol all extracts tested positive for sterols in Salkowaski test. Only chloroform, acetone and methanol extract confirm presence of alkaloids in L. macrophylla roots.

Besides Ethyl acetate and chloroform extract all confirm phenols, however, acetone and methanol extract were found rich in tannins. Petroleum ether and ethyl acetate extract failed to report flavonoids. For anthocyanin confirmation only acetone and aqueous extract tested positive. Anthraquinones was reported from methanol extract only. Chloroform extract only tested positive for cardiac glycosides.  Acetone and methanol extracts of L. macrophylla roots tested positive for maximum phytoconstituents tests.

GC-MS analysis of Leea macrophylla:

Total seven  compounds were identified using GC-MS analysis of methanol extract of L. macrophylla tubers. The results are presented in Table 1.2 while Figure 1 illustrates the chromatogram and the retention times, compounds, peak areas, MWs, and molecular formulae.

       The phyto-compounds listed below were detected by GC-MS investigation of methanolic extract of L. macrophylla tubers viz. Among them are acetamidocinnamic acid; pentadecanoic acid, 14-methyl, methyl ester; cis-2-hexen-1-ol, picolinyloxydimethylsilyl ether; 2-propanol, 1-[1-methyl-2-(2-propenyloxy)ethoxy]; and fensulfothione oxon.

Case study: Effect of Oil extracted from Leea macrophylla Root tubers

A 58 year old male patient who complained of pain due to joint inflammation give the concent for the case study to evaluate the anti-inflammatory property of L.macrophylla. Consent was obtained from the patient and patient asked for a massage technique over the inflammatory area of his foot in the present study. The procedure was done locally for 20 minutes  for seven days then the area was asked to wipe with cotton cloth.

Preparation of oil:

Proportions of three components

Paste of roots: 1 part

Liquid(water): 16 part

Oil: 4 parts

Oil is prepared by crushing the roots in a mortal and pestle and used it afresh if they are taken dry ,a power is used. Paste is boiled in water in the mild fire not closing with lid and by constantly stirring the contents. Then oil is added and it is heated till the total water content is evaporated from the mix. 

Discussion

       Oil extracted from bone healing plants was tried topically to patients who suffering from bone related issues. Topical application of this oil twice a day with gentle massage revealed significant decline in bone pain, and inflammation.

The study justified the anti-inflammatory and wound healing property of Leea macrophylla. Patients treated with oil prepared from bone healing plants of Melghat forest reported positive feedback related to their bone related pains. It also found effective against many arthritis patients with major deformities in bones. Patients suffering from bone loss or brittleness were also showed positive results.

It has been found that the root tuber of L macrophylla contains carbohydrates, proteins, amino acids, lipids, sterols, terpenoids, alkaloids, phenols, tannins, flavonoids, anthocyanins, and anthraquinones. Alkaloids, phenolic chemicals, and steroids were detected in the leaves’ alcoholic extract. Several phytoconstituents, including phenolic chemicals, saponins, glycosides, and flavonoids, were found in the extracts of seeds and leaves after phytochemical screening [8-9]. Potential DPPH, superoxide, and NO free radical scavenging properties are present in L. macrophylla root extracts. The inclusion of oleanolic acid, its derivative 7 α, 28-olean diol, and stigmasterol may be the cause of the plant extracts’ antioxidant properties [10]. Anti-inflammatory, hepatoprotective, antidiabetic, antibacterial, antiurolithiatic, antinociceptive, wound-healing, cytotoxic, antithrombotic, neuroprotective, cardiotonic, and gastroprotective properties are all displayed by the different plant parts in different extracts. Therefore, additional scientific information is needed to investigate its chemical components in the treatment of illnesses and disorders in order to create novel therapeutic medications [11].

 Present results are in support use of L. macrophylla as a valuable source for therapeutic applications by tribal’s of the Melghat forest.

Conflict of Interest: The authors declare no conflict of interest.

References

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