Scientificity of traditional commodity grade of Chinese medicinal materials--taking Lonicerae Japonicae Flos as example / 中国中药杂志

Taking Lonicerae Japonicae Flos as an example, the method of "expert consensus of different regions" was used to screen the representative samples and evaluate their commodity grades. The correlation analysis, hierarchical cluster analysis and partial least squares discriminant analysis(PLS-DA) of "commodity grade-appearance characteristic-component content" were carried out to reveal the scientificity of traditional commodity grade of Chinese medicinal material. By referring to the existing literature and the grade investigation from the sample collection regions, 78 "initial grade" samples were screened out from 118 collected samples. Authoritative experts from four regions(n=4) including Linyi(Shangdong province), Bozhou(Anhui province), Anguo(Hebei province) and Beijing were organized to evaluate their commodity grades, separately. Based on the grade consistency rate(R_i≥70%), 69 "local grade" samples were screened out from the "initial grade" samples. Based on the average grade consistency rate ■ "authoritative grade" samples were screened out from the "local grade" samples, including15 first-grade samples, 9 second-grade samples, 11 third-grade samples and 17 fourth-grade samples. For these "authoritative grade" samples, the main appea-rance characteristics were quantified and the contents of 13 components were determined by ultra performance liquid chromatography(UPLC). Furthermore, the total contents of 6 phenolic acids, 4 flavonoids and 3 iridoids were calculated, respectively. The results of correlation analysis showed that 4 appearance characteristics indices were correlated with the commodity grades: color, rate of yellow bars(including blooming flowers), rate of black heads(including black bars), and rate of stems and leaves(including bud debris). Five component content indices were correlated with the commodity grades: chlorogenic acid, isochlorogenic acid C, sweroside, loganin and the total contents of six phenolic acids. Furthermore, chlorogenic acid, loganin and the total contents of six phenolic acids showed significantly negative correlation with the main appearance characteristics, indicating that the appearance characteristics of Lonicerae Japonicae Flos can reflect its internal quality, and these 3 indices can be used as quality markers(Q-markers). The results of hierarchical cluster analysis showed that the samples of four grades were classified into four categories, and the samples with the same grades and the same categories accounted for 80.8% of the total samples, while the samples with the different grades were obviously classified into different categories. The results of PLS-DA analysis showed that the samples of different grades showed obvious intragroup aggregation and intergroup dispersion. The above results indicated that it was feasible to evaluate the traditional commodity grade of Lonicerae Japonicae Flos by the method of "expert consensus of different regions". For the evaluation of traditional commodity grade of Chinese medicinal material, the samples should be representative, expert conclusions should have enough consensuses, and grade determination should be authoritative. As the crystallization of clinical experience, traditional commodity grade can scientifically reflect the internal quality of Chinese medicinal material.

Nucleosides from Acorus tatarinowii and Their Transformation Pathway / 中国实验方剂学杂志

Objective:To study the water soluble chemical constituents in rhizoma of Acorus tatarinowii and transformation pathway of nucleosides in the process of water extraction. Method:Compounds were isolated and purified by column chromatography on macroporous resin，Sephadex LH-20，ODS and preparative HPLC. Their structures were identified on the basis of physicochemical properties and spectral data. Nucleosides were identified from aqueous extract of A. tatarinowii，and their stability was investigated by HPLC. The possible transformation pathways of nucleosides in aqueous extract of A. tatarinowii were studied by nucleotide addition test. Result:Eleven compounds including four nucleosides，four phenylpropanoids，two alkaloids and a furfural were isolated，and identified as uridine（1），adenine（2），guanosine（3），adenosine（4），5-hydroxymethylfurfural（5），5-（hydroxymethyl）-1H-pyrrole-2-carboxaldehyde（6），（threo）1'，2'-dihydroxyasarone（7），（erythro）1'，2'-dihydroxyasarone（8），acoraminol A（9），acoraminol B（10），and tatarine A（11）. The chromatographic peaks of compounds 1-4 and cytidine were identified from aqueous extract of A. tatarinowii by HPLC. After ultrasonic extraction for 0.5 h，the stability of nucleosides in water was poor. After ultrasonic extraction for 3 h or refluxing extraction for 0.5 h，the stability of nucleosides in water was good. Four transformation pathways including 5'-cytidylic acid→cytidine，uridine monophosphate→uridine，guanosine monophosphate，guanosine and adenosine-5'-monophosphate，adenosine 5'-diphosphate，adenosine 5'-triphosphorate，adenosine，adenine might exist in water extract of A. tatarinowii. Conclusion:Compounds 1-4 and 6 were isolated from the genus Acorus for the first time. These compounds further enriched the chemical constituents of A. tatarinowii. The stability and transformation pathway of nucleosides in A. tatarinowii provides reference data for the analysis of nucleosides in A. tatarinowii and other traditional Chinese medicine.

Variations in the composition of Fructus Evodiae after processing with Radix Glycyrrhizae extract / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the changes in the concentrations of five components in Fructus Evodiae used in Chinese medicine, including evodiamine and glycyrrhizic acid, during processing of Fructus Evodiae with Radix Glycyrrhizae extract by using high performance liquid chromatography (HPLC) and to provide a scientific basis for different clinical uses of processed and unprocessed Fructus Evodiae.</p><p><b>METHODS</b>The concentrations of the Fructus Evodiae components in processed or unprocessed Fructus Evodiae were evaluated by HPLC using a YMC J'sphere ODS-H80 column (4.6 mm×250 mm, 5 μm) with acetonitrile-water-tetrahydrofuran-acetic acid (41:59:1:0.2, v/v/v/v) as the mobile phase. The detection wavelength was 225 nm, the column temperature was 35°C, the flow rate was 1.0 mL/min, and the injection volume was 10 μL. The concentrations of the Radix Glycyrrhizae components were determined by HPLC with a Kromasil-C₁₈ column (4.6 mm×250 mm, 4 μm) and a gradient elution of acetonitrile (A) and 0.05% aqueous phosphoric acid (B) as the mobile phase. The detection wavelength was 237 nm, the column temperature was 35 °C, the flow rate was 1.0 mL/min, and the injection volume was 10 μL.</p><p><b>RESULTS</b>The calibration curves of evodia lactone, evodiamine, rutaecarpine, liquiritin, and glycyrrhizin showed good linear relationships (r>0.99). The recoveries of evodia lactone, evodiamine, rutaecarpine, liquiritin, and glycyrrhizin were 96.59%, 104.18%, 101.91%, 97.75%, and 97.95%, respectively. The concentrations of the components in processed Fructus Evodiae were obviously different to those in unprocessed Fructus Evodiae.</p><p><b>CONCLUSIONS</b>The developed method is rapid and accurate. The results provide a reference for processed Fructus Evodiae and the changes that could be expected in its effects compared to unprocessed Fructus Evodiae.</p>

Evaluation of green tangerine peel processed with vinegar using HPLC fingerprint technique / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for determination of fingerprints and evaluate green tangerine peel (GTP) processed with vinegar.</p><p><b>METHOD</b>The samples were extracted with 50% ethanol using ultrasonic bath and analyzed on a Chromasil C18 column eluted with methanol and water containing 0.2% acetic acid in a linear gradient program. The flow rate was 1.0 mL x min(-1), and detection wavelength was set at 280 nm.</p><p><b>RESULT</b>The RSD values of relative retention times and peak areas of main peaks were all less than 5%. The fingerprints of Geqingpi and Sihuaqingpi were different. The fingerprints of the processed and the unprocessed were almost same, and the fingerprint differences among the different batches processed with the same technology from the same unprocessed GTP were not obvious.</p><p><b>CONCLUSION</b>The influence of processing with vinegar on 50% ethanol extracts were not evident, and the technological pross was stable. It was found that the fingerprints of Geqingpi and Sihuaqingpi were different, but it should be study further.</p>

Effects of different types and standard of processing vinegaron inherent constituents in rhizoma of Corydalis yanhusuo / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects of processing adjuvants-different types of processing vinegar on effective constituents in rhizoma of Corydalis yanhusuo, and evaluate the quality of different types of vinegar in China.</p><p><b>METHOD</b>The HPLC method was adopted to determine the extraction solubility of dl-tetrahydropalmation and total alkaloids in rhizoma of Corydalis yanhusuo processed by vinegar. The sample extracts were separated on kromasil ODS column with mobile phase of methanol-1% phosphoric acid solution(65:35) and detection wavelength was 280 nm.</p><p><b>RESULT</b>There was a remarkable increase in extraction solubility of dl-tetrahydropalmation and total alkaloids in the rhizoma of Corydalis yanhusuo processed by the vinegar products with high content of total acids or with known trademarks.</p><p><b>CONCLUSION</b>Some types of vinegar with known trademarks had been preliminarily selected for the process of Chinese traditional medicine and they also met the requitrement of processing adjuvants with medicine grade. The results will be benefited to the foundation of standardization of vinegar.</p>

Determination of synephrine and N-methyltyramine in Pericarpium Citri Reticulatae viride by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a quantitative method for determination of synephrine and N-methyltyramine in Citri Reticulatae.</p><p><b>METHOD</b>Samples were extracted with 30% methanol. ODS column was used with methanol-water-sodium dodecyl sulfate (55:45:0.1) as mobile phase. Detection wavelength was 285 nm.</p><p><b>RESULT</b>Synephrine and N-methyltyramine in sample solution were well separated. Linearity of synephrine was good (r = 0.9999) in range of 0.35-11.24 microg. The average recovery was 97.1%, and RSD of repeatability was 1.9%.</p><p><b>CONCLUSION</b>This method can be used for quality control of Citri Reticulatae.</p>