Modular hplc system

The dry extract of Galphimia glauca was analyzed in a modular HPLC system (Waters) constituted by a 2695 separation model (Alliance; Waters) and a 2996 photodiode detector (Waters). The equipment was controlled with a data capture computer software program (Empower pro; Waters). The chromatographic method was developed in a reverse-phase column (Alttima, RP- 18, 3 μm, 4.6 × 70 mm; Merck). The mobile phase comprised a 35:65 acetonitrile/water isocratic system eluted at a 1.7 mL/ min flow with a 21 min run time. The fingerprints were obtained at a 220 nm wavelength. For the calibration curve, four ascendant concentrations of G-B which were previously isolated from Galphimia glauca extract were injected in the same chromatographic method (Figure 3). This methodology allowed us to discover that the G. glauca extract contained 53 mg/g of G-B (Figure 1).

The dry extract of Galphimia glauca was analyzed in a modular HPLC system (Waters) constituted of a 2695 separation model (Alliance; Waters) and a 2996 photodiode detector model (Waters). The equipment was controlled with a data-capture computer-software program (Empower 3; Waters). The chromatographic method was developed in a reverse-phase column (Supelcosil RP-18, 5 μm, 4.6 × 250 mm; Merck). The mobile phase consisted of water (solvent A) and acetonitrile (solvent B). The gradient system was as follows: 0-1 min, 0% B; 2-3 min, 5% B, 4-20 min, 30% B; 21-23 min, 50% B 14-15 min; 24-25min, 80% B; 26-27 100% B; 28-30 min, 0% B. The flow rate was maintained at 0.9 mL/min and the injection volume was 10 μL. The fingerprints were obtained at a 230-nm wavelength. For quantitative analysis, previously isolated G-B was used as standard to build a calibration curve. Four ascendant concentrations (0.050, 0.100, 0.200, and 0.400 μg/mL) of this triterpene were injected into the HPLC by triplicated (10 μL). Peak area data obtained at 230 nm allowed obtaining the calibration curve (Tr=27.97 min, R2=0.99) [13 (link)]. This methodology allowed us to know that the G. glauca extract contained 53 mg/g of G-B (Figure 1).