International Research Journal of Plant Science

Gc-Ms analysis, in-vitro neuroprotective and antioxidant studies on ethanolic and acetone extract of oledenlandia corymbosa

Gomathi V¹, Babitha M.C^2*, C. Umamaheswara Reddy³, M.Kumar⁴ and Venkateswarlu B.S^{5 1}Professor, Department of Pharmacology, Vinayaka Mission’s College of Pharmacy, Vinayaka Mission Research Foundation (Deemed to be University), Ariyannur, Salem-636308, India
²Research Scholar, Vinayaka Mission’s college of Pharmacy, Vinayaka mission research foundation (Deemed to be university), Ariyannur, Salem 636308, India
³Deputy Registrar (Invigilation), Sri Ramachandra institute of Higher Education and Research (D.U), Porur, Chennai-600116, India
⁴Professor and Head, Department of Pharmaceutical chemistry, Vinayaka Mission’s College of Pharmacy, Vinayaka mission research foundation (Deemed to be University), Ariyannur, Salem-636308, India
⁵Principal, Vinayaka Mission’s College of Pharmacy, Vinayaka Mission Research Foundation (Deemed to be University), Ariyannur, Salem-636308, India
^*Corresponding Author:
Babitha M.C, Research Scholar, Vinayaka Mission’s college of Pharmacy, Vinayaka mission research foundation (Deemed to be university), Ariyannur, Salem 636308, India, Email: mcbabith@gmail.com

Received: 13-Apr-2022, Manuscript No. IRJPS-22-60606; Editor assigned: 15-Apr-2022, Pre QC No. IRJPS-22-60606(PQ); Reviewed: 29-Apr-2022, QC No. IRJPS-22-60606; Revised: 20-Jun-2022, Manuscript No. IRJPS-22-60606(R); Published: 27-Jun-2022, DOI: http:/dx.doi.org/10.14303/irjps.2022.018

Introduction

Oldenlandia corymbosa Linn.is a flowering plant belonging to the Rubiaceae family and is commonly found in India, tropical East Asia, Java Island and in Srilanka. It is commonly known as Parppatakapullu and is one of the chief ingredients in various ayurvedic preparations like Amritarishtam, Mahatiktaghrtam, Candanasavam, Jatyadi tailam (Das et al., 2019). Oldenlandia corymbosa reported to have immunoprotective activity and used in many traditional medicines to treat ulcers, bronchitis, pelvic and uterine inflammation. Aqueous extract of the plant contains arabinose, rhamnose, mannose, zylose glucose and galactose (Sivapraksam et al., 2014).

Materials and Methods

Plant Material

Whole plant of Oldenlandia corymbosa was collected from Kottayam, Kerala. The plant was identified and authenticated by Dr. M.U. Sharief, The Scientist ‘E’ & Head of office, Botanical survey of India, Southern regional center, Coimbatore. Identification No.: BSI/SRC/5/23/2020/ Tech/63.

Preparation of ethanol and acetone extract

Oldenlandia corymbosa plants (Whole plant) were washed several times with distilled water to remove complete impurities present in it. Then it is dried at room temperature and coarsely powdered and the powder was extracted with ethanol and acetone for 50 hours. Alcohol and acetone are completely removed under reduced pressure, and semisolid extracts were obtained (Redfern et al., 2014).

GC-MS analysis

Shimadzu GC-MS (Model Number: QP2010S) with GC –MS solutions software was used to carry out GC-MS profiling of Oldenlandia corymbosa extract (ethanol and acetone). Chromatographic conditions: Elite-5 MS column( fused silica) of 30 mm length,0.25mm internal diameter and 0.25μ thickness was used. The carrier gas used was Helium at a flow rate of 1 ml /min and the injection volume of the sample was 1.0 microlitre. The oven temperature is maintained at 280°C. The total time taken for GC running was 50 min. By comparing the average peak area to the total area relative percentage amount of each component was calculated.

Invitro Neuroprotective Effect

MTT assay method: In-vitro Neuroprotective potential of selected extract of Oldenlandia Corymbosa (ethanol& acetone) was assessed using IMR-32 Neuroblastoma cells (purchased from NCCS Pune was maintained in Dulbecco’s modified eagles media). After attaining sufficient growth of the cell line Lipopolysaccharide (1 μg/ml) was added to induce neuroinflammation and incubated for one hour, prepared extracts were added to the respective wells. The sample content in the well after 24 hours of incubation period were removed and MTT solution was added to all tests and cell control wells again incubated for 4 hours. After removing the supernatant MTT solubilization solution (DMSO) was added and absorbances were measured by microplate reader at a wavelength of 540 nm (Chang et al., 2001).

In-vitro antioxidant Assay

DPPH Radical Scavenging assays and ABTS assay were used to assess antioxidant potential of the extracts.

DPPH Radical Scavenging assay

Antioxidant activities of ethanol and acetone extracts were determined by DPPH assay. Samples of different concentrations (12.5 μg/mL to 200 μg/mL) mixed with DPPH and this reaction mixture incubated at room temperature in dark condition for 20 minutes, a control without test compound is also prepared. Absorbance was measured at 517 nm and as the positive control Ascorbic acid was used (Rajurkar & Hande, 2011).

ABTS assay

Antioxidant activities of ethanol and acetone were determined by ABTS assay. Extracts of different concentrations mixed with ABTS solution. Generated radical monocation was reduced in the presence of antioxidants present in the extract. Absorbance was measured at 734 nm. Ascorbic acid was used as standard (Massada, 1976).

Results and Discussion

Identification of Components

Detection of phytoconstituents was done using National Institute Standard and Technology (NISTII) and WILEY 8 library. For characterizing phytoconstituents, parameters such as comparison of retention time and peak, computer matching, and the characteristic fragmentation patterns of the mass spectra were used (Adams, 2007; Cazes, 2004; Bhardwaj et al., 2020).

Phytochemicals detected from the chromatogram are shown in Table 1 (ethanol extract) and (Tables 2 and 3) (acetone extract).The chromatograms (Figures 1 and 2) exhibited the presence of numerous biologically active compounds (Lozano et al., 2018).

Table 1. List of Chemical fractions identified in the Ethanol Extract of Oldenlandia Corymbosa.

Table 2. List of Chemical fractions identified in the Acetone Extract of Oldenlandia Corymbosa.

Table 3. Phytocompounds were proven for their antioxidant and anti-inflammatory activity.

In vitro neuroprotective Effects

The viability of cells was evaluated by direct observation of cells by Inverted phase contrast microscope followed by MTT assay. The data obtained are given in the table 4. Percentage viability against concentration graphically represented in figure 3.

Table 4. PercentageViability of cells by MTT assay.

DPPH Radical Scavenging Assay

It is one of most popularly used assay method to detect antioxidant potential of herbal extracts. Table 5 and figure 4 exhibited ethanol extract had got more antioxidant activity compared to ethanol extract of the plant. IC 50 Value from DPPH assay for ethanol extract is 130.56 μg/mL and acetone extract is 189.33 μg/mL.

Table 5. DPPH Assay of extracts of Oldenlandia Corymbosa.

ABTS Assay Method

From the ABTS study also it is clear that ethanol extract has got more antioxidant potential compared to acetone extract. Table 6 and Figure 5. IC 50 Value from ABTS assay for ethanol extract is 1328.38 μg/mL and acetone extract is 1462.17 μg/Ml.

Table 6.  ABTS Assay of extracts of Oldenlandia Corymbosa.

Conclusion

In the present work attempts were made to scientifically validate and standardize the antioxidant and neuroprotective potential of Oldenlandia corymbosa. GC-MS analysis revealed that the plant is rich in phytoconstituents having antioxidant and anti-inflammatory activity. The antioxidant activity against DPPH and ABTS free radicals showed ethanol extract had got more antioxidant activity. Invitro neuroprotective activity against IMR-32 cell lines exhibited by both plant extract this may be due to the presence of antiinflammatory fractions such as neophytadiene, squalene, stigmasterol and thunbergol. Further studies on these plants can result in the development of newer drug entities that can be most efficiently used in neuroinflammation mediated neurodegenerative diseases.

Acknowledgement

The authors thank Department of pharmacology, Vinayaka mission’s college of pharmacy, Yercad main road, Salem for providing the necessary facility to accomplish the work.

Conflict of Interest

The authors declare that there is no conflict of interests regarding the publication of this article.

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Citation: Babitha et al., (2022). Gc-Ms analysis, in-vitro neuroprotective and antioxidant studies on ethanolic and acetone extract of oledenlandia corymbosa. IRJPS. 13: 018.