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A validated high-performance liquid chromatography (HPLC) method has been developed to analyze the (±)-gossypol in the selection of strains of Candida tropicalis culture. Since gossypol was easily degraded and oxidized, the addition of antioxidant NADPH-Na4 and acetone extraction was chosen to prevent gossypol degradation and gradient elution assay was applied to obtain gossypol resolution. Concentrations of gossypol in C. tropicalis ZD-3 culture 20 μg/mL were determined, and concentration–time profiles were observed. Linearity of the gossypol standard curve by HPLC area method was ranged from 0.1 to 20 μg/mL with Y = 26.954 × X − 29.547, R 2 = 0.9991, and n = 3, with limit of detection (LOD) of 50 ng/mL and lower limit of quantification (LLOQ) of 500 ng/mL. The recovery rate is dose-dependent and ranged from 85.3% to 103.5%. It is a rapid and reliable HPLC method for gossypol quantization in microorganism culture which could be applied in solid fermentation in the feed industry.

Open access
Acta Chromatographica
Authors: Mei-Xia Zhu, Sheng-Nan Li, Hai-Dan You, Bin Han, Zhi-Ping Wang, Yan-Xi Hu, Jin Li and Yu-Feng Liu

High-performance liquid chromatography coupled with photodiode array detection and evaporative light scattering detection (HPLC—DAD—ELSD) was established to determine paeoniflorin and albiflorin simultaneously in Radix Paeoniae Rubra. The assay was performed on a Diamonsil C18 (4.6 mm × 250 mm, 5 μm) column by a gradient elution program with acetonitrile and aqueous formic acid (0.05% v/v) as mobile phase at a flow rate of 1.0 mL min−1. The detection wavelength of DAD was 230 nm, and the evaporator tube temperature of ELSD was set at 110 °C with the nebulizing gas flow rate of 3 L min−1. The temperature of column was kept at 30 °C. The linear ranges of paeoniflorin and albiflorin were within 0.050–1.510 mg mL−1 and 1.007–5.035 mg mL−1. The recoveries of paeoniflorin and albiflorin were 96.2–102.9% and 95.0–102.4%, respectively, while the relative standard deviation (RSD) of them was 0.2–2.5%. This method was quick, simple, accurate, and specific. It could be used for the quality control of Radix Paeoniae Rubra. The proposed approach was expected as a powerful tool for the quality control of Radix Paeoniae Rubra.

Open access