ABSTRACT
Objective: To develop an UPLC method for simultaneous determination the contents of 10 active constituents [neochlorogenic acid (NCA), chlorogenic acid (CA), cryptochlorogenic acid (CCA), isochlorogenic acid A (ICAA), isochlorogenic acid B (ICAB), isochlorogenic acid C (ICAC); bacicalin, wogonoside, baicalein, and wogonin] in Yinhuang preparations. Methods: Analysis was performed on an Acquiyt UPLC BEH C18 chromatographic column (100 mm × 2.1 mm, 1.7 μm) eluted with acetonitrile- 0.4% phosphoric acid, gradient elution, and the flow rate was 0.4 mL/min, the detection wavelength was set at 326 nm, the column temperature was 40℃, and the injection volume was 1.0 μL. Results: All calibration curves were linear (r ≥ 0.999 6) over the tested ranges. The average recoveries (n = 9) ranged from 97.43% to 99.94% with RSD value below 2.0%. The contents of 11 batches of NCA, CA, CCA, ICAA, ICAB, ICAC, bacicalin, wogonoside, baicalein, and wogonin were 0.236-3.419, 5.279-26.220, 0.495-4.714, 0.130-2.702, 0.310-3.210, 0.363-5.036, 35.209-133.289, 1.493-6.635, 1.546-5.393, and 0.254-0.823 mg/g. Conclusion: This method is rapid, simple, sensitive, accurate, and reproducible, which can be used for quality control of Yinhuang preparation, and provide a reference for the formulation of quality standards to enhance in the future.
ABSTRACT
This experiment aimed to explore and research the process of preparing baicalein and wogonin through liquid fermentation with Bacillus natto. Active enzymes of produced by B. natto was used for the biological transformation of baclin and wogonoside, in order to increase the content of the haicalein and wogonin in the scutellaria. With the content of the baicalein and wogonin as evaluating indexes, the effects of carbon source, nitrogen source, the types and suitable concentration of inorganic salt, medium pH, granularities of medical materials, liquid volume in flask, shaking speed, liquid-to-solid ratio, fermentation time on the fermentation process were studied. The optimal process conditions for liquid fermentation of scutellaria were 1.0% of peptone, 0.05% of NaCl, pH at 6, the granularities of medical materials of the scutellaria screened through 40-mesh sifter, 33% of liquid, shaker incubator speed at 200 r x min(-1), liquid-to-solid ratio of 5:1, temperature at 37 degrees C, fermentation for 6 days, baclin's conversion rate at 97.6% and wogonoside's conversion rate at 97% in the scutellaria. According to the verification test, the process was stable and feasible, and could provide data reference for the industrial production.