Objective: The objective of this study was to evaluate the nature of chemical constituents, total phenolics, total flavonoids, and antioxidant activity of Cleome gynandra and their functional groups with the help of phytochemical, Fourier-transform infrared spectroscopy (FTIR) analysis, colorimetric assay, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Methods: C. gynandra of the Cleomaceae family is an annual herb. The dried leaves were powdered and extracted using Soxhlet apparatus by different solvents. Preliminary phytochemical analysis was carried out to identify the phytoconstituents present in the extract of C. gynandra, FTIR spectrum was scanned at the range of 4000-400 cm−1. The extracts were subjected to the colorimetric assay in triplicate manner to quantitative determination of total phenolic and total flavonoid content. Gallic acid and rutin used as standards to determine the total phenolic content and total flavonoid content. Antioxidant activity was evaluated using DPPH radical scavenging method. Results: Phytochemical analysis of the ethanolic extract of C. gynandra revealed the presence of alkaloids, phenolics, saponins, steroids, flavonoids, cardiac glycosides, and tannins. FTIR spectrum showed intense bands at 3679.18, 3616.63, 3317.34, 2943.67, 1634.01, 1360.20, 1036.71, and 778.04 cm−1 corresponding to N-H2, O-H stretch, aliphatic C-H stretch, C=O, C-H benzene, C-O stretch, and C-Cl. The total phenolic content was found to be 8.39 ± 0.0952 mg gallic acid equivalent/g and 66.76 ± 0.0333 mg rutin equivalent/g. The DPPH radical scavenging activity of ethanolic extract was showed more scavenging activity compared to ethyl acetate and n-hexane fractions. Conclusion: The present research work creates a platform to screen many bioactive chemical constituents present in C. gynandra to treat various diseases.

A new method was proposed using RP-UPLC for the determination of Gitingensine in bulk, which exhibits its power of stability output. Gitingensine is a natural product found in Kibatalia laurifolia belonging to Apocynaceae which is a steroid having the activities such as anti-inflammatory, anti-spasmodic and anti-cancer activity. Cevadine is used as an internal standard for chromatographic analysis. The elution was performed on BEH C18 (2.1 × 50 mm, 1.7 µm) column at 30 °C with a mobile phase distribution Acetonitrile: 0.1% orthophosphoric acid (60: 40) respectively. The flow rate was well- kept at 0.3 mL min-1. Retention times for Gitingensine and Cevadine were found to be 2.005 and 1. 395 min, respectively. The regression equation was found to be linear in the range of 12.5 – 75 µg/mL with a high correlation coefficient (0.999). Recovery of Gitingensine was obtained as 100.04%. Validation was done as claimed by ICH guidelines with respect to accuracy, sensitivity, robustness, and precision studies. From the insignificant variations in the analysis by changing the mobile phase, temperature, and flow rate, the robustness was studied. All the validation parameters were found to be within the specifications. Forced degradation studies revealed that when the influence of acid, alkali, peroxide, thermal, photolytic, and hydrolytic conditions were applied on the drug, it was stable. Hence, it can be concluded that the developed RP-UPLC method is economical, precise, and robust and can be adopted in regular Quality control analysis.

Antiparkinsonian activity of Cleome gynandra (CG) was evaluated and chemical constituents were isolated. The results of the present study showed that spontaneous motor activity, retention time and number of head drippings were decreased, whereas immobility time in Tail suspension (TST) and Forced swimming test (FST) were increased in MPTP treated animals, while they were significantly (P<0.001) increased and (P<0.01) decreased with various extracts of CG. Dopamine, serotonin (P<0.001), epinephrine (P<0.01) and glutathione levels were significantly decreased and lipid peroxidation was increased in MPTP treated groups, while their levels were significantly (P<0.001) increased and decreased with various extracts of CG in a dose dependant manner as compared to MPTP, L-dopamine and control groups. Ethanolic leaf extract of CG leads to isolation of triterpenoid, flavone and anthocyanidin - O- methylated flavonoids, which were characterized by using spectroscopic investigation methods like FTIR, NMR and MS.