Quantitation and Radical Scavenging Activity Evaluation of Iridoids and Phenylethanoids from the Roots of Phlomis umbrosa (Turcz.) using DPPH Free Radical and DPPH-HPLC Methods, and their Cytotoxicity

The roots of Phlomis umbrosa (Turcz.) (Phlomidis Radix) have been traditionally used to treat cold, reduce swelling and staunch bleeding. Four iridoids (1 – 3 and 5) and six phenylethanoid derivatives (4, and 6 – 10) were isolated from the roots of P. umbrosa. A simple, sensitive, and reliable analytical HPLC/PDA method was developed, validated, and applied to determine 10 marker compounds in Phlomidis Radix. Furthermore, the isolates were evaluated for cytotoxic and anti-oxidant activities as well as DPPH-HPLC method. Among them, compounds 4 and 6 – 9 displayed potent anti-oxidant capacities using DPPH assay with IC50 values of 27.7 ± 2.4, 10.2 ± 1.1, 18.0 ± 0.8, 19.1 ± 0.3, and 19.9 ± 0.6 µM, and compounds 6, 8, and 9 displayed significant cytotoxic activity against HL-60 with IC50 values of 35.4 ± 3.1, 18.6 ± 2.0, and 42.9 ± 3.0 µM, respectively.

Validation of a photostability indicating method for quantification of furanocoumarins from Brosimum gaudichaudii soft extract

ABSTRACT A validation study of a reverse-phase high-performance liquid chromatographic assay for the quantification of two furanocoumarins (psoralen and bergapten) in soft extract obtained from Brosimum gaudichaudii Trécul, Moraceae, roots was conducted. The developed method was sensitive, rapid, reproducible, easy and precise, and showed linearity (r > 0.99) in the range of 10-64 µg/ml for psoralen, and 9-56 µg/ml for bergapten. It also showed a good efficiency for the photodegradation analysis of psoralen and bergapten in the soft extract. The photostability results showed that the Higuchi model presented the best fitting to the obtained data. Both chemical markers showed stability over 2.6 days, suggesting potential applications of the extract in obtaining intermediate products from this plant material. Furanocoumarins take around 30 min to be activated by UV light, reaching the maximum biological potential. Thus, the results obtained to the Higuchi model, corresponding to 2.6 days of stability, shows feasibility with future applications of these chemical markers.

Isolation And Development Of Quantification Method For Cyanidin-3-glucoside And Cyanidin-3-rutinoside In Mulberry Fruit By High-performance Countercurrent Chromatography And High-performance Liquid Chromatography

Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column (4.6 x 250 mm I.D., 5 microm) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with r2 = 0.9996) in the concentration range 15.625 - 500 microg/mL, and the relative standard deviations (RSD%) of intra- and interday variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.