Lc 20 ad prominence liquid chromatograph system

Bouea macrophylla Griffith (Maprang) seeds were collected between March and April 2015 from the Maprang Plantation located in Nakhon Nayok Province. The samples were collected with permission from private landowners and botanically identified by comparison with a voucher specimen (CMUB39942) deposited in the CMUB herbarium at Chiang Mai University, Thailand. The Maprang seed crude extract was prepared, as described previously [13 (link)]. In brief, 50 g of the minced dried seeds were macerated in 75% ethanol (500 mL) for 7 days with daily shaking. Then, the extracts were filtered by Kesselguhr and concentrated by a rotatory evaporator (Buchi Rotavapor R-100, Switzerland). After lyophilization, a total of 20 g of extract powder (MPSE) was obtained (40% yield). MPSE was collected and stored at room temperature in a desiccator for further study. To prepare the stock solution, MPSE was solubilized with deionized water (Pure Lap Option-Q, ELGA, UK) at a final concentration of 1 mg/mL and then filtered with a 0.22 μm syringe filter, aliquoted, and stored at − 80 °C until being used. The active ingredients including, penta-O-galloyl-β-D-glucose hydrate (PGG), ethyl gallate (EG), and gallic acid (GA), were quantified by using Shimadzu LC-20 AD Prominence Liquid Chromatograph system equipped with an SPD-M20A Prominence Diode Array Detector (Shimadzu, Japan).

The stock solution of MPSEs was freshly prepared at 1 mg/mL volume in purified water and was filtrated with a 0.45 μm syringe filter (Whatman's). HPLC profiles of MPSEs were performed using a Shimadzu LC-20AD Prominence Liquid Chromatograph system equipped with a SPD-M20A Prominence Diode Array Detector and with a DGU-20A3 Prominence Degasser (Shimadzu, Japan). The wavelength scanning was done between 190 and 800 nm, while any wavelength occurring at 270 nm was carefully monitored. Notably, 20 μL of each extract (300 μg/mL) was injected into the ZORBAX Eclipse Plus C18 Analytical column (250 × 4.6 mm i.d., 5 μm particle) with an Eclipse Plus-C18 Analytical Guard Column (12.5 × 4.6 mm i.d., 5 μm particle. Successful separation of MPSEs phytochemicals was accomplished using the following mobile phase; mobile A: 0.1% aqueous trifluoroacetic acid and mobile B: 100% acetonitrile. The time program for gradient elution was 0–5 minutes, 5% B; 7–12 minutes, 10% B; 14–19 minutes, 15% B; 21–26 minutes, 20% B; 30–35 minutes, 25% B; 37–45 minutes, 30% B; 50–55 minutes, and 100%B; 60–65 minutes, 5% B.