RESUMO
An HPLC fingerprint and multi-component determination method of Leonurus japonicus was established for comprehensive evaluation and quality control of Leonurus japonicus. The sample was incubated in 70% ethanol in a water bath for 2 h, and the extract was analyzed by HPLC using Kromasil C18 column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.1% formic acid with gradient elution. The flow rate was 1.0 mL·min-1. The temperature of column was 30 ℃. The detection wavelength was 280 nm. HPLC fingerprint of characteristic components of Leonurus japonicus was established. There were 12 common peaks among 25 batches of samples, and 5 of them were identified and determined. Syringic acid, leonurine hydrochloride, rutin, hyperoside or isoquercitrin showed a good linearity in the ranges of 0.426 1-85.22 ng (r = 0.999 9), 7.948-1 590 ng (r = 0.999 3), 10.20-2 040 ng (r = 1.000 0), 2.018-403.6 ng (r = 0.999 9), or 8.704-1 741 ng (r = 0.999 9), respectively. The average recoveries were 99.0%, 97.6%, 97.4%, 96.9% and 98.5% with RSD of 1.1%, 1.8%, 1.4%, 1.5% and 1.3%, respectively. The HPLC characteristic fingerprint of Leonurus japonicus was specific, and this method can simultaneously determine the content of 5 components.
RESUMO
An UPLC method was developed for the studies of fingerprint and quantification of multi-components for Evodiae Fructus. The chromatographic separation was performed on a C₁₈ column (2.1 mm×50 mm,1.7 μm) with mobile phase of 0.2% formic acid-acetonitrile and 0.2% formic acid-water in gradient mode, and the detection wavelength was set at 320 nm．Dehydroevodiamine was used as the reference peak, there were 24 common peaks in the fingerprint of 29 samples were detected, and among them 10 chromatographic peaks were identified with the reference substance and they were neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, hyperin, isorhamnetin-3-O-β-D-rutinoside, dehydroevodiamine, evodiamine, rutaecarpine, evocarpine and dihydroevocarpine. The fingerprint data was evaluated with similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (Version 2008A), and the similarity of 19 batches of Evodiae Fructus was greater than 0.9 in the 29 samples. In addition, 9 components including neochlorogenic acid, chlorogenic acid, hyperin, isorhamnetin-3-O-β-D-rutinoside, dehydroevodiamine, evodiamine, rutaecarpine, evocarpine and dihydroevocarpine were simultaneously determined at the same chromatographic conditions, whose peak area integral values showed good linear relationship at the range of 0.000 46-0.138, 0.000 146-0.175, 0.000 412-0.124, 0.000 448-0.134, 0.000 452-0.136, 0.003 38-0.169, 0.000 44-0.132, 0.001 07-0.128, 0.001 71-0.128, respectively. Their average recoveries were 100.3%, 100.4%, 101.6%, 97.51%, 102.9%, 101.4%, 103.8%, 104.0%, 95.99%, and RSD were 2.4%, 2.0%, 3.0%, 0.80%, 1.9%, 2.1%, 1.1%, 2.2%, 2.4%, respectively. The established UPLC method not only realized the full separation of all chemical constituents of Evodiae Fructus within 20 minutes, but also achieved the chromatographic fingerprint determination and simultaneous multi-components determination of Evodiae Fructus at the same chromatographic conditions. Compared with other methods in literatures, the method has the following characteristics of strong specificity, good separation, high purity of chromatographic peaks, simplity and feasibility, which provides better means for the simultaneous qualitative and quantitative analysis of Evodiae Fructus.
RESUMO
OBJECTIVE: To investigate a rapid approach for quality evaluation of Huoxiang Zhengqi Tincture. METHODS: Hesperidin, glycyrrhizic acid, imperatorin, honokiol, isoimperatorin, magnolol and atractylodin in Huoxiang Zhengqi Tincture were determined by ultra performance liquid chromatography (UPLC) method combined with wavelength switching detection technology. The sample was injected directly without preprocessing. The separation was performed on an ACQUITY UPLC BEH C18 (2.1 mm×100 mm, 1.8 μm) and the column temperature was maitained at 40℃. The mobile phase was composed of acetonitrile and 0.1% phosphoric acid with gradient elution at a flaw rate of 0.3 mL·min-1. Hesperidin was detected at 284 nm; glycyrrhizic acid was detected at 250 nm; imperatorin, honokiol, isoimperatorin and magnolol were detected at 300 nm; atractylodin was detected at 340 nm. RESULTS: The calibration curves of the seven components showed good linearity within their test ranges. The average recoveries for hesperidin, glycyrrhizic acid, imperatorin, honokiol, isoimperatorin, magnolol and atractylodin were 99.3%, 99.5%, 101.5%, 99.3%, 100.6%, 99.0% and 99.6%, respectively. And there were great variations among the contents of glycyrrhizic acid, imperatorin, isoimperatorin and atractylodin in 28 batches of samples from 13 manufactures. CONCLUSION: The proposed method is accurate, simple and rapid, thus providing basis for comprehensive quality control of Huoxiang Zhengqi Tincture.