ABSTRACT
UPLC-Q-TOF-MS and serum pharmacochemistry were employed to study the migrating components in rat sera after intragastric administration of the water extracts of Puerariae Lobatae Radix(PLR) and Puerariae Thomsonii Radix(PTR). After the respective intragastric administration of PLR and PTR extracts, blood samples were collected from the orbital vein. The serum samples were treated by protein precipitation method with methanol and acetonitrile at a ratio of 1∶1 and then passed through Agilent ZORBAX RRHD SB-C_(18) column(3 mm×100 mm, 1.8 μm) and Agilent SB-C_(18) pre-column(3 mm×5 mm, 1.8 μm) with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase. The elution was performed at the flow rate of 0.25 mL·min~(-1), the column temperature of 40 ℃, and the injection volume of 2 μL. By comparison of the total ion chromatogram and secondary fragment ion information of PLR and PTR water extracts, PLR-and PTR-containing sera, and blank serum, we found 42 migrating components(including 17 prototype components and 25 metabolites) in the sera of rats treated with PLR and 35 migrating components(including 15 prototype components and 20 metabolites) in the sera of rats treated with PTR. Thirty-three common components were shared by the two treatments, including 13 prototype components and 20 metabolites. The differences of migrating components in the PLR-and PTR-treated rat sera provide a scientific basis for further study of the active components and quality markers of PLR and PTR.
Subject(s)
Animals , Rats , Drugs, Chinese Herbal , Plant Roots , Pueraria , SerumABSTRACT
This study aims to explore the pharmacodynamic differences of Puerariae Lobatae Radix(PLR), Puerariae Thomsonii Radix(PTR) and their different processed products and the influences of these medical materials on the diversity of intestinal flora. The Sennae Folium-induced diarrhea model, streptozotocin(STZ)-induced diabetes model and L-nitro-arginine methyl ester(L-NAME)-induced hypertension model were used to compare the pharmacodynamic differences in anti-diarrhea, blood glucose reduction and blood pressure lowering among raw, roasted and vinegar-processed PLR and PTR. The effects of raw and processed PLR and PTR on intestinal flora diversity of rats were evaluated by 16 S rDNA high-throughput sequencing. The roasted PLR and PTR performed better in anti-diarrhea, especially the former. PLR and its processed products all presented the efficacy of reducing blood glucose, and the vinegar-processed PLR was the most outstanding. The raw PTR was not that effective in reducing blood glucose, whereas its efficacy was improved after roasting and vinegar processing. Both PLR and PTR were capable of lowering blood pressure to a certain extent, and PLR is superior to PTR in this aspect. Further, the vinegar-processed PLR showed the best effect. The diversity of intestinal flora was different among rats to which different products of PLR and PTR were administered. The roasted PLR led to the highest abundance of Lactobacillus, which was closely related to its best antidiarrheal effect. The highest abilities of vinegar-processed PLR to lower blood glucose and blood pressure were associated with the high abundance of Blautia and Prevotella_9. This study lays a foundation for elucidating the processing mechanisms of PLR and PTR and provides a basis for their further development and application.
Subject(s)
Animals , Rats , Drugs, Chinese Herbal , Gastrointestinal Microbiome , Plant Roots , PuerariaABSTRACT
Objective: To establish HPLC specific chromatograms of Puerariae Lobatae Radix(PLR) and Puerariae Thomsonii Radix(PTR), and make a distinction about their species and different habitats of PLR by chemical pattern recognition, provide reliable methods for scientific evaluation and effective control of their quality. Method: HPLC was employed to determine the contents of chemical ingredients in 23 batches of PLR and PTR.The similarity analyzed with "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica"(version of 2004A), then a common pattern was established.Based on its chemical fingerprint information, the quality of PLR and PTR was comprehensively analyzed by three kinds of chemical pattern recognition methods. Result: In addition to sample S22(from Shaanxi province), the similarities of 23 batches of samples were more than 0.9, which showed that similarity of PLR and PTR was good, this method can not differentiate them.Principal component analysis(PCA) could only identify PLR and PTR, but partial least squares-discriminant analysis(PLS-DA) could distinguish PLR from PTR and the producing areas of PLR with model interpretation of 96.4% and prediction of 74.6%.The result of hierarchical cluster analysis(HCA) was consistent with PLS-DA. Conclusion: Chemical pattern recognition method can make a distinction between PLR and PTR, as well as different habitats of PLR;it is suitable for quality control of their medicinal materials.
ABSTRACT
Objective: To establish the quality control methods for the standard decoction of Puerariae Thomsonii Radix.Method:According to the preparation principles for traditional Chinese medicine (TCM) standard decoction,13 batches of Puerariae Thomsonii Radix from different origins were analyzed under the chromatography conditions established in this study and verified with methodology.By referring to Chinese Pharmacopoeia of 2015,puerarin was used as a quantitative indicator to calculate the transfer rate.In this study,the structures of main chromatographic peaks were also identified to clarify the main chemical constituents in the standard decoction.Result:The 13 batches of medicinal herbs were identified as Puerariae Thomsonii Radix,with a recovery rate of 98.0%,and RSD of 1.1%,indicating that the method was accurate and reliable.The transfer rate ranged from 41.4% to 60.0%,and the extraction rate was within the range of 15.7%-34.3%.The corresponding fingerprints were prepared for 13 batches of the standard decoction,and their similarities were all greater than 90.0%.The chemical constituents from Puerariae Thomsonii Radix were identified by mass spectrometry analysis,including citric acid,4'-O-glucosyl puerarin/daidzein-4',7-diglucoside,3'-hydroxy puerarin/genistein puerarin,puerarin apioside,daidzin puerossid A and daidzein,etc.Conclusion: The 13 batches of Puerariae Thomsonii Radix decoction in different origins had consistent properties with the basic properties of medicinal decoction pieces.The established method of quality evaluation can be used to systematically evaluate the standard decoction,providing reference for quality control of related decoction preparations of Puerariae Thomsonii Radix.
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Objective To establish UV spectrophotometry and HPLC methods for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix and Puerariae Thomsonii Radix; To compare the ultrasonic method at room temperature, conventional refluxing method and ultrasonic method at heating conditions at the aspect of content determinations. Methods The content determinations of total flavonoids was determined by UV spectrophotometry at 250 nm; the content of puerarin was determined by HPLC with octadecylsilane-bonded silica gel as the stationary phase, a mixture of methanol and water (25:75) as the mobile phase, 256 nm as the detection wavelength, 1.0 mL/min as the flow rate. Results Contents of total flavonoids in Puerariae Lobatae Radix by ultrasonic method at room temperature, conventional refluxing method and ultrasonic method were15.09%, 14.48%, and 12.71% (n=3), respectively. The contents of puerarin were 4.37%, 4.09%, and 3.80% (n=3), respectively. Contents of total flavonoids in Puerariae Thomsonii Radix were 2.09%, 2.23%, and 2.17% (n=3), respectively. The contents of puerarin were 0.50%, 0.53%, and 0.52% (n=3), respectively. Conclusion Ultrasonic method at room temperature can replace conventional refluxing method for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix, and ultrasonic method at heating conditions also can replace conventional refluxing method for content determinations of total flavonoids puerarin from Puerariae Thomsonii Radix. Puerarin contents from Puerariae Lobatae Radix and Puerariae Thomsonii Radix in South Anhui Province are all in line with the Pharmacopoeia standards.
ABSTRACT
Objective To establish UV spectrophotometry and HPLC methods for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix and Puerariae Thomsonii Radix; To compare the ultrasonic method at room temperature, conventional refluxing method and ultrasonic method at heating conditions at the aspect of content determinations. Methods The content determinations of total flavonoids was determined by UV spectrophotometry at 250 nm; the content of puerarin was determined by HPLC with octadecylsilane-bonded silica gel as the stationary phase, a mixture of methanol and water (25:75) as the mobile phase, 256 nm as the detection wavelength, 1.0 mL/min as the flow rate. Results Contents of total flavonoids in Puerariae Lobatae Radix by ultrasonic method at room temperature, conventional refluxing method and ultrasonic method were15.09%, 14.48%, and 12.71% (n=3), respectively. The contents of puerarin were 4.37%, 4.09%, and 3.80% (n=3), respectively. Contents of total flavonoids in Puerariae Thomsonii Radix were 2.09%, 2.23%, and 2.17% (n=3), respectively. The contents of puerarin were 0.50%, 0.53%, and 0.52% (n=3), respectively. Conclusion Ultrasonic method at room temperature can replace conventional refluxing method for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix, and ultrasonic method at heating conditions also can replace conventional refluxing method for content determinations of total flavonoids puerarin from Puerariae Thomsonii Radix. Puerarin contents from Puerariae Lobatae Radix and Puerariae Thomsonii Radix in South Anhui Province are all in line with the Pharmacopoeia standards.
ABSTRACT
In order to compare the effect of sulfur fumigation processing and direct hot air heating technology on puerarin contents and efficacy of Puerariae Thomsonii Radix, the fresh roots of Pueraria thomsonii were cut into small pieces and prepared into direct sunshine drying samples, direct hot air drying samples, and sulfur fumigation-hot air drying samples. Moisture contents of the samples were then determined. The puerarin contents of different samples were compared by HPLC method. Moreover, the models of drunkenness mice were established, and then with superoxide dismutase (SOD) content as the index, aqueous decoction extracts of Puerariae Thomsonii Radix samples with sulfur fumigation processing and non-sulfur fumigation processing methods were administrated by ig; the effects of sulfur fumigation on contents of SOD in mice liver and serum were determined, and the sulfur fumigation samples and non-sulfur fumigation samples were investigated for moth and mildew under different packaging and storage conditions. Results showed that the sulfur fumigation samples significantly changed the puerarin content from Puerariae Thomsonii Radix. The content of puerarin was decreased gradually when increasing the times of sulfur fumigation and amount of sulfur. SOD content in drunken mice liver and serum was significantly decreased when increasing the times of sulfur fumigation, showing significant difference with both direct sunshine drying group and direct hot air drying group. Moth and mildew were not found in the sulfur fumigation samples and direct hot air drying samples whose moisture contents were lower than the limit in Pharmacopoeia. Research showed that sulfur fumigation can significantly reduce the content of main active ingredients and reduce the efficacy of Puerariae Thomsonii Radix, indicating that the quality of Puerariae Thomsonii Radix was significantly decreased after sulfur fumigation. However, the contents of the main active ingredients, efficacy and storage results of the direct hot air drying samples were similar to those in direct sunshine drying samples, so the hot air drying process was a nice drying technology which could be promoted for use.