ABSTRACT
Objective: To explore the effect on the accumulation of medicinal compositions β-eudesmol, atractylon, atractylodin and key enzyme genes 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and farnesyl pyrophosphate synthase (FPPS) expression in biosynthesis of Atractylodes lancea under copper stress. Methods: Under copper stress, the expression of key enzyme genes HMGR and FPPS in A. lancea was determined by real-time fluorescence quantitative PCR; the content of three medicinal components in A. lancea were determined by HPLC; The correlation analysis was performed with SPSS, and DPS software for grey correlation analysis. Results: When the copper stress concentration was within 100 mg/kg, the expression of FPPS and the content of atractylon in the rhizomes of A. lancea increased slightly. However, when the copper concentration continued to increase, the expression levels of HMGR and FPPS and three medicinal components content of A. lancea showed a different degrees of downward trend. The expression levels of HMGR and FPPS were positively correlated with the content of β-eudesmol, atractylon, and atractylodin (P < 0.05) under copper stress. Grey relational analysis showed that the content of β-eudesmol and atractylon in the rhizomes was significantly correlated with the expression of HMGR and FPPS of A. lancea under copper stress. The expression of FPPS gene had the larger contribution on the composition of β-eudesmol and atractylon. However, the correlation between the content of atractylodin and the expression of these two key enzyme genes was relatively small. Conclusion: This study clarified the change regulation of two key enzyme gene expression and the content of three medicinal compositions, and revealed the relationship between β-eudesmol, atractylon and HMGR and FPPS, the key enzymes in terpene biosynthesis of A. lancea under copper stress. It contributed to the further study of the molecular regulation mechanism of the synthesis of medicinal constituents under copper stress and provided a theoretical basis for improving the quality of A. lancea.
ABSTRACT
Objective: To study the content determination of atractylodin, atractylon, and β-eudesmol in Atractylodis Rhizoma by GC and characteristic spectrum in order to provide a scientific basis for the quality control. Methods: Using GC and Agilent HP-5 capillary column, taking nitrogen as carrier gas, FID as detector, temperature programming, split ratio, injection port temperature: 250℃, detector temperature: 250℃, column temperature: 130℃; The contents of atractylodin, atractylon and β-eudesmol of 25 samples between Chengde and purchased from other markets were determined by external standard method. The characteristic spectrum was set up and the similarity was analyzed by Estimating System of Similarity on the Chinese Medicine Fingerprint Chromatogram. Results: The determination method and characteristic spectrum by GC for atractylodin, atractylon, and β-eudesmol in Atractylodis Rhizoma were established. Nine characteristic peaks were identified; The linear range of β-eudesmol was 20.00 - 406.10 μg/mL (r = 0.999 9), and the average recovery was 100.75%, RSD = 1.17% (n = 6), and the limit of detection was 0.12 ng. The linear range of atractylon was 35.00 - 348.70 μg/mL (r = 0.999 5), and the average recovery was 99.84%, RSD = 1.29% (n = 6), and the limit of detection was 0.04 ng; The linear range of atractylodin was 16.46 - 329.30 μg/mL (r = 0.999 6), and the average recovery was 100.12%, RSD = 0.88% (n = 6), and the limit of detection was 0.06 ng. Conclusion: The concent determination and characteristic spectrum method of atractylodin, atractylon, and β-eudesmol established by this study are sensitive, simple, stability, which could make the determination result accurate and reliable.
ABSTRACT
A sensitive and selective method based on gas chromatography hyphenated to mass spectrometry (GC-MS) was developed and validated for the determination of atractylon in rat plasma. Plasma samples were processed by liquid-liquid extraction with ethyl acetate-n-hexane (1:1, v/v) using acetophenone as an internal standard (IS). Analytes were determined in selective ion monitoring (SIM) mode using target ions at m/z 108.1 for atractylon and m/z 105.1 for acetophenone. The calibration curve was linear over the concentration range of 10-1000 ng/mL with lower limit of quantification of 10 ng/mL. The intra- and inter-day precision variations were not more than 10.4% and 9.6%, respectively, whilst accuracy values ranged from -6.5% to 4.9%. Extraction recovery of the assay was satisfactory. This method was suc-cessfully applied to quantification and pharmacokinetic study of atractylon in rat plasma after in-tragastric administration of Atractylodis extract.
ABSTRACT
This study was aimed to explore the stability of atractylon in volatile oil of A tractylodes macrocephala Koidz. under various conditions. High performance liquid chromatography (HPLC) method was used to determine the atractylon content and atractylenolide content of volatile oils. And the relative percentages of atractylone and a-tractylenolide were selected as indexes to measure the change of atractylon. The results showed that the atractylon in atractylodes volatile oil was unstable with the rate of atractylon decomp osition up to 12% in natural conditions on the 6th day. The condition was almost not changed at room temperature and avoiding light. Atractylodes volatile oil in tween-80 solution was unstable with the rate of atractylon decomposition up to 30% at room temperature and avoiding light on the 5th day. The condition was almost not changed in the freezing temperature and avoiding light. The atracty-lon in volatile oil in artificial gastric juice and artificial intestinal juice were unstable, which were in conformity with the first order kinetics. It was concluded that the stability of atractylon of volatile oil was different. The atractylon of volatile oil was relatively stable in dilute solution within 48 hours. The atractylon in volatile oil was stable than its pure solution.