1260 series high performance liquid chromatograph

An Agilent 1260 series high-performance liquid chromatograph, equipped with a quaternary pump, UV detection, an autosampler, a column temperature controller, and a vacuum degasser (Agilent Technologies, Palo Alto, CA, USA) was used in the HPLC assay of A. rupestris. The samples were separated using a Hypersil GOLD C₁₈ column (5 μm, 250 × 4.6 mm). The injection volume of sample and standards were all 10 μL, and the column temperature was maintained at 35 °C. A mixture of acetonitrile (A) and 0.2% phosphate solution (B) was used as the mobile phase at a flow rate of 1.0 mL/min. The gradient elution mode was modified as follows: 0–15 min, 5–18% A; 30 min, 20% A; 35 min, 21% A; 40 min, 40% A; 47 min, 45% A; 51–55 min, 80% A; 55.1–65.0 min, 5% A. The detection wavelength was sat at 350 nm.

The Agilent 1260 series high-performance liquid chromatograph (Palo Alto, CA, USA) equipped with a quaternary pump, autosampler, warm column chamber and a diode array detector (DAD) was used to perform HPLC analysis of the Oviductus Ranae sample. The column model used was Agilent TC-C18 (250 × 4.6 mm, 5 µm), and the injection volume was 10 µL. The flow rate was set to a linear gradient of 1.0–0.5 mL/min for 0–14 min and maintained at 0.5 mL/min for 14–30 min. The gradient elution conditions of the mobile phase were 0–10 min linear gradient 86–93% A, 10–20 min maintain 93% A, 20–30 min linear gradient 93–100% A. The mobile phase A was acetonitrile, and the composition of mobile phase B (water or 1% phosphoric acid aqueous solution) was optimized according to the separation effect of the chromatographic peak. The detection wavelength of the sample was confirmed according to the DAD full-wavelength scan result and the column temperature was optimized to achieve the best separation of the chromatographic peaks. Agilent Chemstation software (Palo Alto, CA, USA) was used to record and process the acquired HPLC data.