We developed a method to analyze the fingerprint spectrum qualitatively and quantitatively for the traditional Chinese herbal medicinal preparation Gan-Lu-Yin with HPLC combined with photodiode array detection, and MS, and to identify the preparation's 14 main components including baicalin, baicalein, oroxylin A-7-O-glucuronide, wogonin-7-O-glucuronide, wogonin, and oroxylin A in Radix Scutellariae; naringin and neohesperidin in Aurantii fructus; liquiritigenin, liquiritin, and glycyrrhizic acid in Radix Glycyrrhizae. In LC/UV assay, a Cosmosil 5C18-MS-II column was used as the stationary phase, and a gradient of potassium dihydrogen phosphate, ACN, and water as the elute solution. The UV detection wavelengths were 250 and 280 nm. In LC/MS assay, a gradient of phosphoric acid, ACN, and water was used as the elute solution, and electrospray positive ion mode ((+)-ESI) as the analytic mode. In order to explore the distribution of trace metal elements effectively in Gan-Lu-Yin, a microwave digestion method was used for sample treatment, and an inductively coupled plasma MS assay was used to analyze fingerprint spectra of the inorganic metals in Gan-Lu-Yin. Combined with fingerprint spectra of organic compounds by LC/UV and LC/MS, it was expected to provide effective quality control in the production of Gan-Lu-Yin.
A facile high-performance liquid chromatography (HPLC) method for the resolution and quantitative measurement of nine marker substances, the active ingredients in patch preparations of Ru-Yi-Jin-Huang-San, was established using gradient elution in the reversed-phase mode. These marker substances included berberine (Phellodendri Cortex), curcumin (Curcumae Rhizoma), imperatorin (Angelicae Dahuricae Radix), magnolol (Magnoliae Cortex), hesperidin (Citri Leiocarpae Exocarpium), glycyrrhizin (Glycyrrhizae Radix), and emodin, sennoside A, sennoside B (Rhei Rhizoma). The ingredients in the water-based and oil-based patches of the formula from different manufactures were also analyzed for quality evaluation. Extracted samples were analyzed by HPLC using a reversed-phase column (Inertsil 5 ODS-2, 4.6-mm I.D. x 250 mm) at 30 degrees C and eluted with a mixture of 20 and 70% acetonitrile aqueous solution in gradient manner at a flow rate of 1.0 ml/min. The detection wavelength varied with time as follows: 275 nm, 0-72 min; 250 nm, 72-105 min; 220 nm, 105-145 min. Relative coefficients of variations of intra- and interday analysis were less than 5%. All the recoveries were 93.30-113.63%. This method could be applied for the simultaneous determination of nine marker substances in Ru-Yi-Jin-Huang-San.
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