Spd m20a

Analytical HPLC-DAD analysis was performed using Shimadzu HPLC equipment (Shimadzu, Tokyo, Japan) coupled with an automatic degasser (DGU-20A 3R), a quaternary pump (LC-20AD), an auto-sampler (SIL-20A HT), a DAD detector (SPD-M20A), and an Agilent Zorbax Eclipse XDB-C18 (250 mm × 4.6 mm, 5 µm) column. Water (A) and methanol (B) with the following gradient was used: 0 min, 50% B; 5 min, 60% B; 25 min, 80% B; and 30–35 min 100% B. The flow rate was 1 mL/min; the column temperature was 25°C, the injection volume was 10 μL and the detection wavelength was set at 254 nm.
All purified compounds, identified as 6′,7′-dihydroxybergamottin (1), officinalin (2), stenocarpin isobutyrate (3), officinalin isobutyrate (4), 8-methoxypeucedanin (5), and peucedanin (6), were dissolved in methanol (0.1 mg/mL) and the stock solutions were diluted in a series of appropriate concentrations (20–100 μg/mL) for the construction of calibration curves. The results of the quantitative analysis were expressed as mean ± standard deviation mg compound/100 g of dry weight plant material. The LOD and LOQ values were calculated from the standard deviation of y-intercepts (σ) and the slopes (S), according to the following formulas: LOD = 3.3 × σ/S and LOQ = 10 × σ/S. All experiments were repeated three times.

The contents of 5’-nucleotides of different treatments were analyzed using dried mushroom powders by HPLC analysis using a 254 nm diode array detector (Prominence SPD-M20A) and a C18 column (250 mm × 4.6 mm, 5 μm, Agilent, USA) at a flow rate of 1.0 mL/min. The injection volume was 20 μL, and the column temperature was 30 °C. The mobile phase consisted of 50 mmol/L phosphate buffer and methanol (97: 3, v/v) [16 (link)]. Each 5’-nucleotide was identified using the authentic 5’-nucleotide (Shanghai Anpu Experimental Technology Co., Ltd., Shanghai, China).

5-HMF determination was performed by HPLC [37 (link)] using a Shimadzu SPD-M20A diode array detector and an Agilent ZORBAX SB-C18 column (5 μm, 9.4 × 250 mm). The mobile phase consisted of water/methanol (90:10, v/v) at a flow rate of 1.0 mL/min. The column temperature was 40 °C and a 10-μL sample was injected into the HPLC system. The UV detection wavelength was set at 285 nm. Data analysis was performed using LC Solution software.