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Objective: To compare the differences of Gardeniae Fructus and its stir-baked preparation and screen out the differential markers, and provide reference for the establishment of quality standard. Methods: The whole and partial fingerprints of Gardeniae Fructus and its stir-baked prepared slices were established by HPLC-UV analysis. And the fingerprints were statistically analyzed using hierarchical clustering analysis (HCA), principle component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA), then the characteristic markers were screened out. And then content of 6-α-hydroxygeniposide, gentioside, geniposide, crocin I, and crocin II was determined. Results: A total of 15 peaks were acquired for Gardeniae Fructus and stir-baked Gardeniae Fructus respectively, 12 peaks of which were the common peaks. The similarity of whole and partial reference fingerprints of Gardeniae Fructus and its stir-baked prepared slices was 0.991 and 0.880, respectively, and partial fingerprint can successfully distinguished Gardeniae Fructus from its stir-baked prepared slices. HCA and OPLS-DA results showed that significant differences were observed between Gardeniae Fructus and stir-baked Gardeniae Fructus with peak 3, peak 5, peak 9 (geniposide), and peak 11 (crocin I), which being identified as the main differential markers. Contents of geniposide, crocin I and crocin II in Gardeniae Fructus were higher than those in stir-baked Gardeniae Fructus, while 6-α-hydroxygeniposide was lower. Conclusion: The composition was significantly different before and after stir-baked. The established partial fingerprints combined with multi-component determination method can effectively distinguish them. And the differential markers obtained by multivariate statistical analysis can provide reference for the selection of quality markers.
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Objective: To evaluate the effect of irradiation of 60Co-γ on composition change of Curcuma Longa through HPLC fingerprint analysis and determination of curcumin content. Methods: An Agilent SB-C18 column (250 mm × 4.6 mm, 5 μm) was used as the column. Curcumin content was measured according to the method in 2015 edition of the “Pharmacopoeia of the People’s Republic of China”. Acetonitrile-0.1% phosphoric acid aqueous solution was used as the mobile phase of fingerprint analysis, and it was programmed in a gradient elution model and the detection wavelength was 240 nm. The effects of irradiation on content of curcumin and components change were analyzed by paired t-test and similarity evaluation. And then the comprehensive evaluation of constituent difference of C. Longa before and after irradiation was carried out by HCA and PCA analysis. Further more the stability of curcumin content and fingerprint similarity of three patches samples storage for 3 and 6 months was compared with that of 0 month. Results: There was no significant difference before and after irradiation for curcumin content (P > 0.05). The fingerprints of C. Longae were established and 16 peaks were identified as the common peaks. The similarity of each batch before and after irradiation was more than 0.998 and the same batch before and after irradiation have better clustering consistency. The changes of curcumin content for 3 and 6 months compared with 0 month was less than 5%, and the fingerprint similarity was all greater than 0.95. Conclusion: The established fingerprinting method is accurate and reliable. 60Co-γ irradiation has no significant effect on the curcumin content and chemical composition consistency of C. Longa, which did not affect its stability. It was a good way to evaluate the 60Co-γ irradiation effect on C. Longa by combining fingerprint analysis and the index components content determination. This method can provide reference for the utilization of irradiation on C. Longa.
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Objective: To establish the HPLC fingerprint of Angong Niuhuang Pills (ANP) and determine the contents of six compounds with analysis of their quality by chemical pattern recognition. Methods: The separation was performed with acetonitrile-0.1% phosphoric acid aqueous solution as the mobile phase, and the detection wavelength was set at 230 nm. Twelve batches of samples were collected from four different companies and analyzed using the evaluation system of traditional Chinese medicine chromatographic fingerprint similarity, and chemometric Methods:. The content of the main effective components of geniposidic acid, geniposide, berberine hydrochloride, baicalin, wogonosidc, and wogonin were determined. The samples of different companies were analyzed by chemical pattern recognition. Results: The similarity of fingerprints was above 0.9, the content of six kinds of main effective components showed that there were some differences in the 12 batches of products, especially the composition of Scutellaria baicalensis. The results indicated that the quality of S. baicalensis was not stable. Among them, the contents of baicalin, wogonosidc and wogonin of company were more stable than other companies, and even higher than other companies. The samples can be sorted according to their quality through the F score of the PCA. The results showed that the samples of C company in the comprehensive ranking ranked more forward, which were consistent with the quantitative results. Conclusion: Fingerprints combined with chemical pattern recognition can comprehensively response the quality of ANP, providing the experiments basis of reasonable controlling the quality of preparation.
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AIM To determine the contents of five marker components and total flavonoids in Paishi Granules (a medication to remove stones in the body,containing Glechomae Herba,Akebiae Caulis,Cynanchi Paniculati Radix et Rhizoma,etc.).METHODS The content determination of various components in 50％ methanol extract of Paishi Granules was accomplished by RP-HPLC.The analysis was performed on a 30 ℃ thermostatic Aglient SBC1s column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.05％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 240 nm.The content of total flavonoids was detected by ultraviolet spectrophotometry.RESULTS Chlorogenic acid,loganin,rutin,rosmarinic acid and ammoninm glycyrrhizinate showed good linear relationships within the ranges of 18.88-604.00,18.50-592.00,20.25-648.00,18.75-600.00 and 18.81-602.00 μg/mL,whose average recoveries were 97.9％,100.7％,103.3％,96.4％ and 102.4％ with theRSDsof1.4％,0.4％,2.7％,1.2％ and 2.3％,respectively.The contents of total flavonoids in twenty batches of samples all met pharmacopeia requirements,while those of marker components were found to be different,among which loganin displayed the most significant change,followed by rutin and ammoninm glycyrrhizinate.CONCLUSION This simple,accurate and reliable method can be used for the quality control of Paishi Granules.
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To compare the intestinal absorption and metabolism of icariin in different osteoporosis rat models. Ovariectomy and intragastric administration of cyclophosphamide were used to establish two kinds of rat osteoporosis models. Then the rat intestinal perfusion was conducted, and HPLC was used to measure and calculate the permeability coefficients of icariin in different intestines and production amount of metabolites. Western blot was used to detect LPH enzyme expression in two models. Experimental results showed that both ovariectomy and intragastric administration of cyclophosphamide 4.5 mg•kg⁻¹ could reduce rat bone density and successfully construct the rat osteoporosis models. The apparent permeability coefficient Papp of 20 μmol icariin in duodenum, jejunum, ileum, colon was 5.695, 5.224, 1.492, 0.520 respectively in sham operation group; 3.876, 3.608, 0.863, and 0.291 in ovariectomized group; 4.945, 3.601, 1.990, 1.042 in normal saline group; 3.301, 2.108, 1.209, 1.233 in cyclophosphamide-induced osteoporosis model group. In addition, the protein expression levels of LPH enzyme in two model groups were lower than those in normal group. The absorption and metabolism of icariin in two kinds of osteoporosis models was lower than that in sham operation group and normal saline group; the reduction of expression level of LPH enzymes in rat intestine of different osteoporosis models was one of the reasons for leading to the reduced intestinal absorption and metabolism of icariin.