Quantification of Enhydrin and Uvedalin in the Ethanolic Extract of Smallanthus sonchifolius Leaves Validated by the HPLC Method.

Yacon leaf (Smallanthus sonchifolius, Asteraceae) ethanolic extracts are widely used in herbal medicine preparation for diabetes. They contain two sesquiterpene lactones (enhydrin (1) and uvedalin (2)) as major bioactive compounds. To provide a suitable method of analysis for the extract's quality control, we developed and validated a simultaneous HPLC-UV method using the compounds as markers. Compounds 1 and 2 were isolated using a freeze crystallization technique followed by a preparative HPLC. Spectrometry data for 1 and 2 were determined and compared to the literature. Chromatographic separation was carried out for 30 min with a mobile phase that used 60% water and 40% acetonitrile and a C18 column (250 × 4.6 mm, 5 µm) as the stationary phase. The flow was set to 1 mL min-1 and detection was conducted at 210 nm. The validation method was conducted according to the ICH guidelines, which included linearity, precision, accuracy, LOD, and LOQ. The calibration curve of both compounds was linear (R 2 > 0.9999), with the limit of detection and quantification as follows, respectively, 0.52 and 1.57 µg/mL for 1, and 0.144 and 0.436 µg/mL for 2. The percentages of recovery and repeatability (%RSD) were, 101.46 and 0.30% for 1, and 97.68 and 0.08% for 2, respectively. The 1 and 2 were 1.67 and 0.88% in the Ykal extract, and 1.26 and 0.56% in the Ycin extract, respectively. The method was found to be linear, precise, accurate, and suitable to be applied for control quality analyses of yacon leaf extract.

TLC-Based Fingerprinting Analysis of the Geographical Variation of Melastoma malabathricum in Inland and Archipelago Regions: A Rapid and Easy-to-Use Tool for Field Metabolomics Studies.

Melastoma malabathricum is an Indo-Pacific herb that has been used traditionally to treat numerous ailments such as wounds, dysentery, diarrhea, toothache, and diabetes. The objective of this study was to evaluate the variability of the metabolic profiles of M. malabathricum across its geographic distribution. By employing thin layer chromatography (TLC), specimens collected from six terrestrial and archipelago regions of Indonesia were analyzed by densitometry for metabolomic fingerprinting analysis combined with chemometric tools: principal component analysis (PCA) and hierarchical cluster analysis (HCA). Two PCAs were identified as PC1 and PC2 with 41.90% and 20.36%, respectively. Our results indicate the importance of considering geographic distribution during field-collection efforts since they demonstrate regional metabolic variation in secondary metabolites of M. malabathricum, as illustrated by TLC and their biological activities.

Isocratic high-performance liquid chromatography (HPLC) for simultaneous quantification of curcumin and piperine in a microparticle formulation containing Curcuma longa and Piper nigrum.

Poor bioavailability has been reported as a major challenge in the development of curcumin as a pharmaceutical agent. However, co-administration of curcumin with piperine has been shown to improve curcumin bioavailability. Therefore, to assure product control quality, an analytical method needs to be developed for the determination of curcumin and piperine content in a dosage form formulation. The objective of this study was to develop a simple isocratic reversed-phase HPLC (RP-HPLC) method to simultaneously quantify curcumin and piperine content in solid dispersion based microparticle formulation containing Curcuma longa and Piper nigrum extracts. The method was validated according to the International Council for Harmonization (ICH) guideline. Chromatographic separation of three curcuminoids and piperine could be achieved using acetonitrile-methanol-water of 65:5:35 %, at a flow rate of 1 mL/min and a wavelength of 353 nm for detection. Resolution (Rs) of 3.57 and 1.68 for piperine and curcumin, respectively, a theoretical plate number (N) > 8000 and a tailing factor (T) < 1.5 indicate a satisfactory separation of the compounds. The calibration curve was linear from 1.25-15 µg/mL and 2.5-30 µg/mL for piperine and curcumin, respectively, with the correlation coefficient of >0.999. The intra-day/inter-day accuracy and precision demonstrated a recovery of 99.54-101.50%/99.38-99.89% and 100.78-102.51%/101.15-102.47% with a Relative Standard Deviation (RSD) of 0.53-0.95%/0.13-1.44 % and 0.28-1.62%/0.46-1.14% for piperine/curcumin. The limit of detection (LOD) were 0.27 and 0.42 µg/mL, for piperine and curcumin, which reveals an adequate sensitivity. A solid dispersion based microparticle formulation containing C. longa and P. nigrum extracts confirmed the validity of the developed method as a recovery of 91.14% and 99.14% for piperine and curcumin, respectively. In conclusion, all the tested parameters confirm the precision, accuracy, and reliability of the method for the simultaneous analysis of curcumin and piperine within a microparticle formulation containing C. longa and P. nigrum extracts.

Anti-Proliferative Activity and Apoptosis Induction of an Ethanolic Extract of Boesenbergia pandurata (Roxb.) Schlecht. against HeLa and Vero Cell Lines.

Rhizomes of Boesenbergia pandurata (Roxb.) Schlecht have been reported to contain active compounds with anticancer properties. This research was carried out to examine anti-proliferative and apoptotic induction against HeLa and Vero cells-line. Dried powder of B. pandurata rhizomes was extracted by a maceration method using 90% ethanol. Cytotoxic assays to determine IC50 and anti-proliferative effects were carried out by MTT methods. Observation of apoptosis was achieved with double staining using acridine orange and ethidium bromide. The results showed that ethanolic extract of B. pandurata was more cytotoxic against HeLa cells (IC50 of 60 µg/ mL) than Vero cells (IC50 of 125 µg/mL). The extract had higher anti-proliferative activity as well as apoptotic induction in HeLa than Vero cells. Therefore, it was concluded that the ethanolic extract of B. pandurata had anti-proliferative as well as apoptosis induction activity dependent on the cell type.

Saponins from Plumeria acuminata Ait induce cell growth inhibition and apoptosis of oral squamous carcinoma cells.

OBJECTIVE: to investigate the cytotoxic and apoptotic effects of saponins from Plumeria acuminata Ait on oral squamous carcinoma cells (OSCC). METHODS: OSCC cells seeded at 2 × 104 cells/well in a 96-well plate were treated with saponins from P. acuminata Ait and cisplatin in various concentrations for 24 h. Trypan blue dye exclusion assay and ethidium bromide/acridine orange were used to evaluate their cytotoxic and apoptotic effects on the cells, respectively. RESULTS: the results showed that both saponins and cisplatin had cytotoxic and apoptotic effects on OSCC cells. CONCLUSION: saponins from P. acuminata Ait may be potential anti-cancer agents for OSCC.