Study on high throughput sequencing identification of Fructus Arctii and five counterfeit species mix power / 中国中药杂志

Fructus Arctii is a traditional Chinese medicine. The main counterfeit species are the seeds of Arctium tomentosum, Onopordum acanthium, Silybum marianum, Saussurea costus, Amorpha fruticosa. Traditional identification methods or molecular barcoding techniques can identify Fructus Arctii and its counterfeit species. However, the identification of the mixture of it and its spurious species is rarely reported. In this paper, we sequenced the ITS2 sequences of Fructus Arctii and 5 kinds of spurious species mix powder by high-throughput sequencing to identify the mixed powder species and providing new ideas for the identification of Fructus Arctii mix powder. The total DNA in mixed powder was extracted, and the ITS2 sequences in total DNA was amplified. Paired-end sequencing was performed on the DNA fragment of the community using the Illumina MiSeq platform. The sequence was analyzed by the software FLASH, QIIME and GraPhlAn etc. The results showed that the high quality ITS2 sequences of 39910 mix samples were obtained from the mixed samples, of which the total ITS2 sequence of the samples genus was 34 935. Phylogenetic analysis showed that the samples contained Fructus Arctii, A. tomentosum, O. acanthium, S. marianum, S. costus and A. fruticosa. Using ITS2 sequences as DNA barcodes, high-throughput sequencing technology can be used to detect the Fructus Arctii and its spurious specie in mixed powder, which can provide reference for the quality control, safe use of medicinal materials of Fructus Arctii and the identification of mixed powder of traditional Chinese medicine.

Qualitative and quantitative analysis of major constituents of raw and processed Arctii Fructus by UHPLC-UV-Q-TOF-MS/MS / 药学学报

In this study, we developed a qualitative analytical method based on liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) for identification of multi-constituents of raw Fructus Arctii (RFA) and processed Fructus Arctii (PFA). We established a UHPLC-UV analytical method for simultaneously determining 6 major compounds in Fructus Arctii. UHPLC- Q-TOF-MS/MS qualitative analysis was performed under negative and positive ion modes and a total of 23 chemical compounds were identified. The analysis data were subjected to a principle component analysis with a t-test. Ten peaks were found to be the main difference (P<0.05) between RFA and PFA. HPLC-UV quantitative method result showed the contents of 6 constituents were different between RFA and PFA. The results indicated that there was less arctiin, chlorogenic acid, isochlorogenic acid A in PFA than in RFA. However, there were higher levels of arctigenin, isochlorogenic acid B, isochlorogenic acid C in the PFA than RFA, which may be the main reason for different clinical efficacy of RFA and PFA.

Study on the Quality Standard of Fructus arctii Concentrated Granules / 中国药房

OBJECTIVE:To establish the quality standard of Fructus arctii concentrated granules. METHODS:TLC was con-ducted for the qualitative identification and HPLC was used for the content determination of arctiin in F. arctii concentrated gran-ules. The determination was performed on Dionex C18 column with mobile phase consisted of methanol-water(40∶60,V/V) at the flow rate of 1.0 ml/min. The detection wavelength was set at 280 nm,and the column temperature was 30 ℃. The samples size was 10 μl. RESULTS:TLC spectrum showed that the spots of F. arctii concentrated granules and arctiin reference medicinal material had the same color;the linear range of arctiin was 0.5-12.5 μg(r=0.999 8)with the average recovery of 98.41%(RSD=0.81%, n=9). The RSDs of precision,stability,repeatability tests were no more than 1.0%. CONCLUSIONS:The method is feasible and reproducible,and can effectively control the quality of F. arctii concentrated granules.

Optimization of the processing technology of Fructus Arctii by response surface methodology / 中国天然药物

The present study was designed to optimize the processing of Fructus Arctii by response surface methodology (RSM). Based on single factor studies, a three-variable, three-level Box-Behnken design (BBD) was used to monitor the effects of independent variables, including processing temperature and time, on the dependent variables. Response surfaces and contour plots of the contents of total lignans, chlorogenic acid, arctiin, and arctigenin were obtained through ultraviolet and visible (UV-Vis) monitoring and high performance liquid chromatography (HPLC). Fructus Arctii should be processed under heating in a pot at 311 °C, medicine at 119 °C for 123s with flipping frequently. The experimental values under the optimized processing technology were consistent with the predicted values. In conclusion, RSM is an effective method to optimize the processing of traditional Chinese medicine (TCM).

Factors of ultrasonic-assisted extraction of arctiin and arctigenin from Fructus arctii / 中成药

AIM: To investigate and optimize the technology of ultrasonic-assisted extraction of arctiin and arctigenin from Fructus arctii.METHODS: The effects of ultrasonic power,particle size,ratio of liquid to solid,extraction time,extraction temperature and duty factor on the extraction rates of arctiin and arctigenin were investigated,and optimized with orthogonal experiments.RESULTS: The results showed that the optimal extraction conditions were as follows: particle 80 ～ 100 mesh 14 mL/g,ratio of liquid to solid 400 W ultrasonic power,50 ℃ of extraction temperature,and 10 min of extraction time.CONCLUSION: The optimized conditions are reasonable and reliable.

Study on the IR fingerprint of Chinese medicinal granule / 中成药

Objective: To identify the Chinese medicinal granule (CMG) by measuring IR fingerprints. Methods : 12 species drugs were extracted with butanone respectively and then the obtained extracts were measured by the FT-IR spectrometer. Results : By IR fingerprint of 12 kinds of CMG, we found that different batches of the same CMG had a stable and repeatable fingerprint. Conclusion : By using IR fingerprint, CMG can be identified. It provides a rapid monitoring for drug identification and quality control.

Research of Fructus Arctii and its extracts on the renal pathological changes of STZ-induced diabetic rats / 中成药

AIM: To investigate the influence of Fructus Arctii and its extracts to the renal pathological changes of diabetic rat and its protective mechansim METHODS: The rat model induced by 55mg/kg STZ once intraperitoneally was adoped as the diabetic nephropathy of early stage, the extract and the powder of Fructus Arctii were given to the diabetic rat for six weeks. We observed the changes of general condition, and examined the expressions of TGF-? 1 (transforming growth factor-? 1) mRNA and MCP-1mRNA (monocyte chemoattractant protein-1) by reverse transcription-polymerase chain reaction (RT-PCR). CONCLUSION: The ethanol extract of Fructus Arctii relieved renal pathological changes of diabetic rat. The mechanism perhaps was related to the reduction in the expression of MCP-1mRNA and TGF-? 1mRNA in kidney; The study proved that the ethanol extract of Fructus Arctii was more effective than its water extract and coarse powder.

Adsorption and separation properties of AB-8 resin for purifying arctinin / 第三军医大学学报

Objective To study the separation ability of AB-8 macroporous resin in the purification of arctinin in Fructus arctii. Methods HPLC was used to measure the content of arctinin, and the adsorption performance and the elution parameters were investigated. Results The optimal separation conditions were as follows: the concentration of Arctinin was 5.5 mg/ml with a flow rate of 2 BV/h, and 50% alcohol was used as eluant. The adsorption of Arctinin was 52.08 mg/g, and the elution ratio of arctinin was 93.8%, and the purity of arctinin reached 65.2%. Conclusion AB-8 resin can be used to refine the arctinin in the extraction of Fructus arctii.