Reversed Phase High-Performance Liquid Chromatographic Ultra-violet (Photo Diode Array) Quantification of Oleanolic Acid and its Isomer Ursolic Acid for Phytochemical Comparison and Pharmacological Evaluation of Four Leucas Species Used in Ayurveda.

CONTENT: Different Leucas species are well known as "Dronpushpi," a well-known herb of Ayurveda, used in the treatment of various ailments. OBJECTIVE: Evaluation of four industrially important Leucas species for their in vitro antidiabetic potential and radical scavenging effect along with high-performance liquid chromatographic quantification of the bioactive triterpenes. MATERIALS AND METHODS: The quantification of triterpenes was carried out on C-18 column with acetonitrile and water (90:10) as the solvent system at a detection wavelength of 210 nm. In vitro antidiabetic activity was evaluated by α-amylase inhibition assay based on starch-iodine and 3,5 dinitrosalicylic acid (DNS) method. Antioxidant activity was calculated by five different models, namely total phenolic and total flavonoid content, free radical scavenging activity by 1-1-diphenyl-2-pic-rylhydrazyl (DPPH), ferric-reducing power assay, and the total antioxidant capacity. RESULTS: Maximum concentration of oleanolic acid was found in Leucas cristata, followed by Leucas mollissima, Leucas Aspera, and Leucas biflora. Ursolic acid was highest in L. mollissima and then in L. biflora, L. cristata, and L. aspera, respectively. In in vitro antidiabetic activity, IC50 of L. aspera (1.56 ± 0.01 mg/ml) and L. mollissima (0.75 ± 0.005 mg/ml) were found to be highest in DNS and iodine starch assay. IC50 in DPPH assay ranges from 0.6 ± 0.011 to 1.68 ± 0.011 mg/ml. Antioxidant capacity follows the order; L. aspera > L. mollissima > L. biflora > L. cristata. CONCLUSION: Promising activities were observed in targeted species, thus L. mollissima, L. biflora, and L. cristata can be used alternatively as a substitute to L. aspera. SUMMARY: Physicochemical parameters are within the limit as per the Ayurvedic Pharmacopoeia of IndiaMaximum concentration of oleanolic acid was found in Leucas cristata; however, ursolic acid was highest in Leucas mollissima In vitro antidiabetic activity of Leucas aspera and L. mollissima was found to be heighest as compared to other species. However, antioxidant capacity is almost similar in targeted species.Promising activities were observed in all the species, thus L. mollissima, Leucas biflora, and L. cristata can be used alternatively as a substitute to L. aspera.

High-performance thin-layer chromatography densitometric method for the simultaneous determination of three phenolic acids in Syzygium aromaticum (L.) Merr. & Perry.

A simple, precise, and rapid high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous determination of 3 phenolic acids, i.e., gallic acid, caffeic acid, and syringic acid, in the dried buds of Syzygium aromaticum, commonly known as clove. HPTLC was performed on silica gel 60F254 plates with toluene-ethyl acetate-formic acid (8 + 2 + 1) mobile phase and densitometric scanning at 280 nm. The method was validated for selectivity, linearity, precision, and repeatability. Instrumental precision coefficient of variation (CV) was 0.88, 0.93, and 0.98% and repeatability of the method (CV) was 0.76, 0.64, and 0.69% for gallic acid, caffeic acid, and syringic acid, respectively. The linear concentration ranges were 400-3200 ng/spot with a correlation coefficient of 0.993 for gallic acid, 440-3520 ng/spot with a correlation coefficient of 0.994 for caffeic acid, and 400-4000 ng/spot with a correlation coefficient of 0.993 for syringic acid. The average recoveries of gallic acid, caffeic acid, and syringic acid were 96.3, 95.7, and 92.4%, respectively. Gallic acid, caffeic acid, and syringic acid were present at levels of 1.58, 0.06, and 0.05% (w/w), respectively, in S. aromaticum. This method is simple, accurate, precise, and economical and can be used for routine quality control.

Optimization of a high-performance liquid chromatography method for the separation and identification of six different classes of phenolics.

Flavonoids are chemical moieties widely distributed in certain plants that are important biologically active constituents of a daily human diet, with significant pharmacological potential (anti-hepatotoxic, anti-allergic, anti-inflammatory, anti-osteoporotic, and anti-tumor activities). Thus keeping in view the importance of this class of compounds, a rapid method for the separation and identification of fifteen phenols belonging to six different types of phenolics in a sole analysis has been developed and validated using selectivity, precision, recovery, and robustness as parameters. The method developed, which is rapid, accurate, and robust for the analysis of different classes of phenols, can be used in the quality control and standardization of plant extracts as well as herbal drugs, including compound herbal formulations.