Improvement of Quality Standard for Mongolian Medicine Juniperus rigida / 中国药房

OBJECTIVE：To provide scientifi c evidence for improving the quality standard of Mongolian medicine Juniperus rigida. METHODS ：Totally 10 batches of J. rigida from different places were taken as samples to observe their characters and identify them by microscope ；TLC method was adopted to qualitatively identify isoquercitrin ，quercitrin，amentoflavone， podocarpusflavone A and hinokiflavone ；the contents of total ash ，acid-insoluble ash ，ethanol-soluble extract and heavy metals were determined by related method stated in 2020 edition of Chinese Pharmacopeia （part Ⅳ）. The contents of above 5 components in samples were determined by HPLC. RESULTS ：The powder of J. rigida was green or yellowish green ，polygonal tracheids ， closely arranged in longitudinal with unequal stomatal ；epidermal cells were nearly rectangular ；sclerenchyma cells were quasi rectangular and the wall beadedly thickening. Results of TLC showed that the spots of the same color were found in the corresponding positions of chromatogram for test sample and substance control. The contents of total ash ，acid-insoluble ash and ethanol-soluble extract in 10 batches of samples were 7.37％-11.18％，0.75％-2.98％，16.55％-26.42％，respectively；average contents were 8.51％，1.27％，22.35％. The contents of lead ，arsenic，cadmium，mercury and copper were 2.00-5.44，0.65-1.65， 0.044-0.100，0.034-0.160，4.59-6.79 mg/kg，respectively；average conte nts were 3.73，0.97，0.078，0.061，5.23 mg/kg. The linear ranges of isoquercitrin ，quercitrin，amentoflavone，podocarpus- flavone A and hinokiflavone were 4.98-20.02，49.99-199.96， 19.94-99.96，9.99-40.00，20.20-159.98 μg/mL（all r＞0.999 7）； com RSDs of precision ，repeatability and stability （24 h） tests were all less than 3.00％（n＝6）；the average recoveries were 话：0993-2057878。E-mail：Tanghuishz@qq.com 100.62％-102.96％，RSDs were 1.21％-1.88％（n＝6）. Average contents of the above-mentioned 5 compounds in 10 batches of samples were 0.089-0.379，1.379-4.250，1.077-2.026，0.162-0.423， 0.016 9-0.117 0 mg/g，respectively. CONCLUSIONS ：The qualitative and quantitative analysis methods of Mongolian medicine J. rigida are established. It is preliminarily proposed that the total ash content shall not exceed 10.22％，the acid-insoluble ash content shall not exceed 1.53％，ethanol-soluble extract content shall not be less than 17.88％，heavy metal lead should not exceed 5 mg/kg，arsenic should not exceed 2 mg/kg，cadmium should not exceed 0.3 mg/kg，mercury should not exceed 0.2 mg/kg，copper should not exceed 20 mg/kg.

One step assay of the four plant ingredients in Yangxue Anshen syrup by HPLC method / 药学实践杂志

Objective To establish a HPLC method for simultaneous determination of quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in Yangxue Anshen syrup. Methods Waters symmetry C18 column (250 mm×4.6 mm, 5 μm) was used with 0.1% acetic acid (A) and methanol (B) as the mobile phase. Gradient elution was performed at a flow rate of 1.0 ml/min, 0-15 min, 95%-90%A; 15-35 min, 90%-70%A; 35-55 min, 70%-60%A; 55-85 min, 60%-50%A; 85-95 min, 10%A. The detection wavelengths were 256 nm and 320 nm. Column temperature was 30 ℃ and the injection volume was 10 μl. Results Quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside showed good linear relationship within the range of 10-300, 5.0-150.0, 5.0-150.0, 20.0-600.0 µg/ml(r≥0.9989), respectively. The average recovery was (96.75±1.41)%, (99.61±1.01)%, (97.18±1.96)% and(99.12±0.97)% (n=6), respectively. Conclusion The established method is simple, accurate and stable, which can be used for the simultaneous determination of 4 components in Yangxue Anshen syrup.

Quality evaluation of Cyclocarya paliurus from different habitats by fingerprint and multi-component quantification combined with chemical pattern recognition / 中草药

Objective: Based on the method of UPLC fingerprint, multi-component quantification and chemical pattern recognition, the quality of Cyclocarya paliurus from different producing areas was evaluated to provide basis for further development and utilization. Methods: The UPLC fingerprints of 20 batches of C. paliurus from different habitats in five provinces were established to determine the common peaks. Three chemical components were identified and the content of the samples was determined by comparison with the reference materials. Cluster analysis and principal component analysis were carried out by SPSS 22.2 and SIMCA software. Results: A total of 16 chromatographic peaks were selected as the common peaks of the fingerprint, 20 batches of C. paliurus could be divided into six categories by cluster analysis, and the results of principal component analysis and cluster analysis were basically the same; By principal component analysis, the cumulative variance contribution rate of the five principal component factors was 86.765%. The quality of S14 and S15 samples from Enshi, Hubei Province was the best, and S4, S5, S8, and S16 of Xiushui, Jiangxi Province were the second. The content of isoquercetin, quercitrin and afzelin were 0.360%-0.884%, 0.263%-1.097%, and 0.092%-0.403%, respectively, among which the total content of S4, S8, S14, and S15 were the top 4, and that of S18 and S20 was in the last two places. Conclusion: Combined with fingerprint, cluster analysis and principal component analysis, we can evaluate the quality of C. paliurus more comprehensively. Isoquercetin, quercitrin and afzelin can be used as the index components for C. paliurus quality control.

Determination of Six Active Ingredients in Gualoupi Injection by Quantitative Analysis of Multi-components by Single Marker / 中国药师

Objective: To develop a method of quantitative analysis of multi-components by single marker (QAMS) for six active ingredients in Gualoupi injection. Methods: An HPLC method was used with a Waters Sunfire ODS C18column (250 mm×4. 6mm, 2. 5 μm), the mobile phase was methanol(A)-0. 2% glacial acetic acid solution(B) with gradient elution, the flow rate was 1. 0 ml· min-1,the detection wavelength was 254 nm,the column temperature was 30℃ and the injection volume was 20 μl. Using 3, 29-dibenzoyl rarounitriol as a reference, the relative correction factors among karounidiol, vanillic acid, adenosine, quercitrin and cynaro-side were detected by QAMS and their contents were calculated, and the results were compared with those of the external standard method. Results: The differences were not statistically significant between the calculated values of karounidiol, vanillic acid, adeno-sine, quercitrin and cynaroside and those measured by the external standard method (P>0. 05). Conclusion: QAMS can be used for the determination of 6 effective components in Gualoupi injection, and the result is accurate, simple and effective.

Molecular mechanism of quercitrin on osteogenic differentiation and adipogenic differentiation of rat bone marrow stromal stem cells (rBMSCs) / 中草药·英文版

Objective: The study was designed to investigate the molecular mechanism of quercitrin on osteogenic differentiation and adipogenic differentiation of rBMSCs. Methods: rBMSCs were harvested from SD rats, and determination of alkaline phosphatase (ALP) activity, quantification of mineralization by Alizarin Red S staining, and the mRNA expression of osteogenic differentiation markers (Runx2, BMP-2, and OSX) by RT-PCR after rBMSCs stimulated by osteogenic induction with (0.1–10) µg/mL of quercitrin, quantification of Lipid droplet by Oil Red O staining and the mRNA expression of adipogenic differentiation marker (PPARγ C/EBPα and aP2) by RT-PCR after rBMSCs stimulated by adipogenic induction with (0.1-10) µg/mL of quercitrin. Results: Quercitrin can up-regulate the mRNA expression of osteogenic differentiation markers (Runx2, BMP-2, and OSX) and increase ALP activity and mineralization after osteogenic induction, on the other hand quercitrin can suppress the mRNA expression of adipogenic differentiation markers (PPARγ C/EBPα and aP2) and decrease lipid droplet after adipogenic induction. Conclusion: This study suggested that quercitrin not only stimulated osteogenic differentiation but also inhibited adipogenic differentiation of rBMSCs, which was associated with the up-regulation of Runx2, BMP-2, and OSX mRNA expression and the down-regulation of PPARγ C/EBPα and aP2 mRNA expression.

Pharmacokinetics of rutin, quercitrin, and isoquercitrin in Hedyotis diffusa in normal and tumor-bearing rats / 中草药

Objective: Based on the tumor-bearing rat model, using rutin, quercitrin, and isoquercitrin from Hedyotis diffusa as the research object, the pharmacokinetics of those three flavonoid glycosides in the pathological state was studied. Methods: Establishing a method for the comparison of the pharmacokinetics of flavonoid extracts in normal rats and tumor-bearing rats, which was analyzed by HPLC-MS/MS in subcutaneous tumor models of SD rat made with tumour cell of Walker-256. Results: Compared with the pharmacokinetics parameters of flavonoid glycosides in normal rats, the Cmax and AUC0-∞ of rutin, quercetin, and isoquercitrin in the tumor-bearing rats were significantly decreased, t1/2z was prolonged, and the metabolic time of three components was prolonged to 24 h, which revealed the effect of pathological condition on the pharmacokinetic characteristics of flavonoid glycosides. Conclusion: The method established in this study is simple, fast, sensitive, and suitable for the pharmacokinetic study of flavonoid glycosides in rats in vivo. The pharmacokinetic characteristics of flavonoids in normal and tumor-bearing rats are different.

Spectrum-effect relationship of anti-inflammatory effect of different part from Polygonum capitatum / 中草药

Objective To study the spectrum effect relationship between the water and ethanol extracts of Polygonum capitatum and its anti-inflammatory activity. Methods The inflammatory model of mice RAW264.7 cells induced by lipopolysaccharide (LPS) was established, and the release of inflammatory factors was detected by NO and TNF-α kit. The relationship of chemical constituents of P. capitatum was established by using the partial least square method to associate the fingerprint data established by UPLC-MS method and the pharmacodynamics index of different extracts of P. capitatum. Results Different extracts of P. capitatum had no cytotoxicity in the range of concentration less than 250 mg/L, and all of them had anti-inflammatory effect, which had a dose-effect relationship. By using partial least square method, the correlation degree between TNF-α and chromatographic peak was compared. The results showed that the correlation among the peaks 24, 17, 22, 23, 20 and the anti-inflammatory effect was positive, and the anti-inflammatory effect was negatively correlated with the peaks 11, 1, 7, 15, 5, 3. From the analysis of the five peaks, the results showed that all the four peaks were flavonoids except for the tannic acid at peak 17. Conclusion Different extracts of P. capitatum inhibited the inflammatory response of RAW264.7 cells. The flavonoids in P. capitatum had significant anti-inflammatory activity. The results showed that quercetin, tannic acid, and hyperoside had great contribution to the anti-inflammatory effect of P. capitatum.

Effect of P-glycoprotein on the transmembrane absorption and metabolism of quercitrin on Caco-2 cells / 国际药学研究杂志

Objective To explore the role of P-glycoprotein(P-gp)on cellular uptake and metabolism in the transmembrane transport of quercitrin.Methods Caco-2 cell monolayer and P-gp inhibitor Cyclosporin A(CysA)were used in the study.Quercitrin, quercetin,isorhamnetin and tamarixetin were determined by LC-MS to study cellular uptake and metabolism of quercitrin on Caco-2 cells.Results Uptake of quercitrin by Caco-2 cells:in different concentration groups of quercitrin coincubating with and without Cy?sA,intracellular accumulation presented the following characteristics:the amount of quercitrin first rose,reached the peak in 60 min and then declined to a steady-state in 120 min.And meanwhile there were significant differences between the two different processing groups incubating with and without CysA(P<0.05);quercetin was detected in all groups(3.0,9.0 and 27.0 mg/L).But in the higher concentration groups incubating with and without CysA,the intracellular quercetin presented a characteristics similar to its original glycosides and showed a significant difference(P<0.05),while the other groups showed no concentration-and time-dependence.At the same time,isorhamnetin and tamarixetin were detected in two higher concentration groups incubating with and without CysA, which showed the trend similar to the original glycosides but no significant difference was obtained between the two processing groups(P>0.05).Isorhamnetin and tamarixetin were not detected in the low and middle concentration groups.Transmembrane transport:on the basal lateral of all groups,the content of the quercitrin in 150 min incubation time showed a trend of continuous rise,and there was no significant difference between the two processing groups.Quercetin,isorhamnetin and tamarixetin were not detected.Conclu?sion Intact quercitrin could be absorbed into the Caco-2 cells and transported across the Caco-2 cell monolayer,and suffered a series of further metabolism in the Caco-2 cells and the basal side of Caco-2 cell monolayer,leading to different characteristics between intra?cellular accumulation and transmembrane transport.P-gp reduces the transmembrane transport of quercitrin by its efflux function,but did not involved in quercitrin metabolism.

Content analysis of avicularin,quercitrin and quercetin in Herba Taxilli from different host plants / 国际药学研究杂志

Objective To study the content difference of avicularin,quercitrin and quercetin in Herba Taxilli from different host plants. Methods The contents of avicularin,quercitrin and quercetin in Herba Taxilli were determined by RP-HPLC and the samples were prepared by ultrasonic extraction with methanol. The analytical column was UltImate XB-C18(250 mm×4.6 mm,5μm), the mobile phase was acetonitrile-methanol-0.1%phosphate with gradient elution,at a flow rate of 1.0 ml/min. The column tempera-ture was set at 25℃,the detection wavelength was 254 nm for avicularin and quercitrin,and 365 nm for quercetin. Results The lin-ear range of the above three ingrediets were 0.0992-1.9840(r=0.9999),0.2254-4.4580(r=0.9999)and 0.1258-2.5160μg(r=0.9997) and the average recovery rates(n=5)were 98.39%,97.08%and 98.159%,respectively. Their contents from different host plants were 0.0000-0.0398,0.3977-0.7639 and 0.0068-0.0231 mg/g in branches and 0.0167-0.1704,1.8626-11.0041,and 0.0185-0.1841 mg/g in leaves. Conclusion The method is accurate,simple and reproducible,and can be used for the quality control of Herba Taxilli.

Determination of six flavonoids in Schizonepeta tenuifolia from different areas by HPLC / 中草药

Objective: To detect the contents of six kinds of flavonoids in Schizonepeta tenuifolia from different areas, so as to provide a basis for their quality control. Methods: Ultra performance liquid chromatography (UPLC) was used to establish the detection scheme of flavonoids content, and clustering analysis was conducted by the results of content detection. Results: The contents of six kinds of flavonoids in S. tenuifolia from different areas had large differences. The linear ranges were: 0.030-0.148 μg for cynaroside, 0.222-1.108 μg for quercetin, 0.357-1.784 μg for hesperidin, 0.058-0.292 μg for luteolin, 0.054-0.269 μg for apigenin, and 0.050-0.247 μg for diosmetin, respectively. The average recovery rate was between 96.32% and 99.97%, RSD < 2.20%; Among this, the Schizonepeta Briq. herb from Anhui was as one class, the herbs from Henan 1 and Guangdong were as one class, and the herbs from Henan 2, Hebei, Yunnan, Sichuan, Hubei, and Gansu were as the other class. Conclusion: The method used for quality control of the multi-index level in this experiment is simple and sensitive; on the other hand, the herbs from Henan 1, Henan 2, Hebei, Yunnan, Sichuan, Hubei, Guangdong and Gansu have a close relationship; They can be used as the same medicinal source.

Application of QAMS in quality control of Wuzi Yanzong Pills / 中草药

Objective To establish a quantitative analysis of multi-component with a single-marker (QAMS) method for the quality control of Wuzi Yanzong Pills (WYP). Methods Six main effective components (schisandrin, hyperin, quercitrin, kaempferol 3-O-rutinoside, deoxyschizandrin, and γ-schizandrin) of WYP were simultaneously separated on a reversed-phase column (Ultimate LP-C18) with high-resolution of each chromatographic peak by high performance liquid chromatography (HPLC). Schisandrin was selected as the internal reference, and the relative correlation factors (RCFs) of other five components were calculated to achieve QAMS. The ruggedness of RCFs was tested on different instruments and columns. Moreover, results of the QAMS were compared with the external standard method. Results Within a certain linear range, the RCFs of hyperin, quercitrin, kaempferol 3-rutinoside, deoxyschizandrin, and γ-schizandrin were 0.36, 4.86, 0.88, 7.34, and 6.35, respectively. The repeatability was good under different experimental conditions. There were no significant differences between the calculated value and estimated value on QAMS and external standard method. Conclusion The QAMS method can be used to assay the content of six components of WYP simultaneously and control the quality of WYP simplely, reliably, and accurately.

Variation analysis of flavonoids cotents and nitric oxide production inhibition in different parts from Hypericum perforatum / 中草药

Objective To provide a chemometric analytical approach for different parts classification from Hypericum perforatum using ultra performance liquid chromatography (UPLC) combined with chemometrics methods. Methods The ACQUITY UPLC® BEH C18 (50 mm × 2.1 mm, 1.7 μm) column was used, and 0.2% formic acid aqueous solution-acetonitrile as gradient elution system. The chromatograms information of different parts that including flower, fruit, leaf, stem and root from Hypericum perforatum L. was collected. The original data were pretreated by centralization and normalization, analyzed by partial least squares discriminant analysis (PLS-DA) and PLS-tree cluster analysis, and monitored by the half inhibition concentratiom of nitric oxide (NO) production activity as an anti-inflammatory factor, in order to evaluate the similarities and differences in flavonoids cotents and nitric oxide production inhibition in different parts from H. perforatum. Results All the calibration curves of 6 flavonoids showed good linearity in each range with correlation coefficients greater than 0.999 that had good precision, repeatability and stability, and the average recovery ranged from 97.28% to 102.84%. By PLS-DA and PLS-tree, according to the data of flavonoids contents and NO product inhibition activities, it showed the quality of leaf > flower > stem > fruit > root for H. perforatum. Conclusion The established method suggested that an appropriate harvest part of H. perforatum is the stem part on the ground.

Study on fingerprint of Saxifraga stolonifera herbs and determination of five components by HPLC / 中草药

Objective To establish a method to research the fingerprint and determine the ingredients of Saxifraga stolonifera by HPLC, which aimed at providing reference for the quality control of Saxifraga stolonifera. Methods Agilent Eclipse XDB-C18 (150 mm × 4.6 mm, 5 μm) column was used as the stationary phase, and the mobile phase consisted of acetonitrile (A)-0.1% phosphoric acid with gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was 254 nm, and the column temperature was maintained at 35 ℃. The result would be analyzed by SOP of Similarity evaluation system for chromatographic fingerprint of TCM. Results The results obtained by the method of fingerprint analysis are good. In the fingerprints, 11 peaks were selected as the common peaks to evaluate the similarities among 21 batches of S. stolonifera collected from different regions, and five contents of them were indentified as protocatechuic acid, gallic acid, bergenin, quercetin-5-O-β-D-glucoside, and quercitrin. The similarities between standard herb and each determined herb showed a lot of differences from others. The determination method showed that there weregood linear relationships of five figured contents in the range of 0.052 8-0.844 8, 0.020 96-0.335 36, 0.241 6-3.865 6, 0.130 8-2.092 8, and 0.023 68-0.378 88 μg. Moreover, the recoveries rates of five figured contents are 96.64%, 100.72%, 96.62%, 103.71%, 96.75%,and all RSDs were lower than 2%. The contents of 5 components in the gathered herbs were determined by external standard method in the range of 0.07-0.40, 0.19-4.36, 1.42-5.98, 0.42-6.86, and 0.11-1.51 mg/g. Conclusion The method established in this study is simple, reliable and durable, which can provide a scientific basis for the quality control of S. stolonifera.

Analysis of metabolites of quercitrin in rat intestinal flora by using UPLC-ESI-Q-TOF-MS/MS / 中国中药杂志

To investigate the metabolism of quercitrin in rat intestinal flora and possible biological pathways, laying the foundation for the metabolic mechanism of traditional Chinese medicine glycosides ingredients. UPLC-Q-TOF-MS/MS method was established to detect the quercitrin and its metabolites with 0.1% formic acid solution(A)-0.1% formic acid acetonitrile(B) as the mobile phase for gradient elution at a flow rate of 0.3 mL•min⁻¹. Electrospray negative ion mode was applied to analyze the metabolites of quercitrin in rat intestinal flora. Metabolite ToolsTM, mass defect filter(MDF) and other technologies were used to screen, analyze the metabolites and infer the chemical formula of the metabolites. The results showed that quercitrin would have degalactoside, deoxygenation and acetylation reactions, and the aglycone quercetin resulted from degalactoside would have further reactions such as hydroxylation, deoxygenation, reduction, and ring opening to achieve deoxygenation metabolite kaempferol, C2-C3 double bonds hydrogenation and reduction product taxifolin, and degalactoside product quercetin. The research results showed that quercitrin can be metabolized by rat intestinal flora, which could increase their hydrophobicity and chemical diversity.

Study on the Metabolism of Hedyotis Diffusa Flavonoid Extract by Intestinal Bacteria in Vitro / 医药导报

Objective To study and compare the metabolism of Hedyotis diffusa flavonoid extract in intestinal flora from human, normal rats and pseudo germ-free rats in vitro. Methods Intestinal flora of human, normal rats and pseudo germ-free rats were incubated with Hedyotis diffusa extract, rutosid, isoquercitrin and quercitrin under anaerobic conditions at 37℃.High-performance liquid chromatography with diode-array detection ( HPLC-DAD ) was used for the analysis of components and metabolic products.The trends of metabolic transformation of the various components were analyzed according to the changes in chromatographic peak areas at different incubation time points. Results The components of Hedyotis diffusa extract and its monomer were quickly metabolized by human and normal rat intestinal flora and the main metabolite was quercetin.However, they were hardly metabolized by intestinal flora from pseudo germ-free rats. The metabolism effect of intestinal flora was weaker on components than on the corresponding monomers. Conclusion By comparing flavonoids metabolism in intestinal flora from human, normal rats and pseudo germ-free rats, the important role of intestinal microflora in the metabolism of flavonoids is further confirmed.

Simultaneous Determination of Three Flavonoids in Tianjihuang Granules by HPLC / 中国药师

Objective: To establish an HPLC method for the simultaneous determination of three flavonoids in Tianjihuang gran -ules.Methods:A ZORBAX Eclipse XDB-C18 column was used.The mobile phase was methanol-0.2%phosphoric acid solution (42∶58).The detection wavelength was 350 nm and the flow rate was 1.0 ml· min-1.Results:The linear range of isoquercitrin, quer-citrin and quercetin was 4.572-114.300 μg· ml-1(r=0.999 8), 7.028-175.700 μg· ml-1(r=0.999 8) and 2.195-54.880 μg· ml-1(r=0.999 9), respectively.The average recovery was 98.01%(RSD=1.24%, n=6), 98.14%(RSD=0.98%, n=6) and 97.00%(RSD=1.09%, n=6) , respectively.Conclusion:The method is specific and simple,which can be used for the quality con-trol of Tianjihuang granules.

UPLC fingerprint of Viciae Amoenae Herba / 中草药

Objective: To establish the ultra performance liquid chromatography (UPLC) fingerprint of Vicia amoena var. angusta and compare the other five different origin varieties of Viciae Amoenae Herba in order to evaluate the quality of the preparation. Methods: Eclipse Plus C18 RRHD column (50 mm×2.1 mm, 1.8 μm) were used with acetonitrile (B) and 0.1% formic acid aqueous solution (A) in gradient elution mode at a flow rate of 0.2 mL/min. The detection wavelength was set at 340 nm and the column was 30℃. The fingerprints for V. amoena var. angusta from 10 different areas were set up, the similarity assay for V. amoena var. angusta from ten different areas was carried out to evaluate their quality by Chinese Materia Medica (CMM) Fingerprint Similarity Evaluation System (2004A edition), and the peaks were identified by comparing the reference substance with LC-MS/MS; Whilie the other origins of five kinds of varieties of Viciae Amoenae Herba were compared and identified in the same condition. Results: UPLC fingerprints for V. amoena var. angusta were established, six chromatographic peaks among 15 common peaks were identified by UPLC-ESI-Q-TOF-MS and be intended to be chlorogenic acid, myricitrin, hyperoside, isoquercitrin, kaempferit-rin, and quercitrin. UPLC fingerprint mutual pattern of other five kinds of origins varieties of Viciae Amoenae Herba and V. amoena var. angusta had obvious differences. Conclusion: This method is simple, rapid, stable, and reproducb1e, and could be used for the full quality evaluation of Viciae Amoenae Herba.

Chemical constituents from stems and leaves of Cirsium henryi / 中草药

Objective: To study the chemical constituents from the stems and leaves of Cirsium henryi. Methods: The chemical constituents were isolated by various chromatography techniques and their structures were elucidated on the basis of spectroscopic analysis. Results: Sixteen compounds were isolated and identified to be stearic acid (1), 2,3-dihydroxypropyl hexadecanoate (2), palmitic acid (3), taraxasterol (4), pseudo taraxasterol (5), taraxa-sterol acetate (6), β-sitosterol (7), daucosterol (8), protocatechuic acid (9), uracil (10), linarin (11), apigenin (12), quercitrin (13), acacetin (14), acacetin-7-O-β-D-glucoside (15), and 4-hydroxy- 3,5-dimethoxy benzoic acid (16), respectively. Conclusion Compounds 3 and 10-16 are isolated from the plant for the first time, and compounds 10, 13, and 16 are isolated from the plants of Cirsium Mill. Emend. Scop. for the first time.

Isolamento químico e validação analítica por cromatografia líquida de alta eficiência de quercitrina em Solidago chilensis Meyen (Asteraceae) / Chemical isolation and analytical validation by high-performance liquid chromatography of quercitrin in Solidago chilensis Meyen (Asteraceae)

RESUMO A espécie Solidago chilensis Meyen, Asteraceae é conhecida como erva-lanceta ou arnica-brasileira, sendo utilizada popularmente como antimicrobiana e para o tratamento de inflamações tópicas. No entanto, estudos fitoquímicos e farmacológicos para as partes aéreas são escassos. Neste trabalho, realizou-se a determinação de flavonoides por espectrofotometria de UV/Vis, prospecção fitoquímica da fração acetato de etila visando o isolamento do constituinte químico majoritário e validação analítica por cromatografia líquida de alta eficiência (CLAE). O teor de flavonoides totais foi de 5,42%, representados como hiperosídeo. O fracionamento químico utilizando métodos cromatográficos (cromatografia líquida em coluna gel de sílica; CHCl3:EtOH; 8:2 v/v) e espectroscópicos (1H RMN,13C RMN e ESI-MS) revelou o isolamento de quercetina-3-O-α-L-ramnosídeo(quercitrina). A sensibilidade e a linearidade (r = 0,999) da validação analítica, utilizando a quercitrina isolada do extrato hidroalcoólico da planta, revelaram um rendimento de 5,29% do analito em relação à droga vegetal. Precisão, recuperação e robustez, além dos valores estabelecidos para os limites de detecção (LOD) e de quantificação (LOQ), poderão ser utilizados como parâmetros de qualidade para extratos à base de S. chilensis.

ABSTRACT The species Solidago chilensis Meyen Asteraceae, known as “erva-lanceta” or “Brazilian arnica”, is popularly used as an antimicrobial and topical treatment for inflammations. However, phytochemical and pharmacological studies of its aerial parts are scarce. In this study, flavonoids were determined by UV/Vis spectrophotometry and phytochemical screening of the ethyl acetate fraction with the goal of isolating the major chemical constituent and analytically validating it through high performance liquid chromatography (HPLC). The total flavonoid content was 5.42%, represented as hyperoside. Chemical fractionation using chromatographic (liquid chromatography in column of silica gel, CHCl3:EtOH, 8:2 v/v) and spectroscopic methods (1H RMN, 13C RMN, and ESI-MS) revealed the isolation of quercetin-3-O-α-L-rhamnoside (quercitrin). The sensitivity and linearity (r = 0.999) using the isolated quercitrin of the hydroalcoholic extract of the plant revealed a yield of 5.29% of analyte in relation to the plant. Precision, recovery, and robustness, as well as values set for the limits of detection (LOD) and quantitation (LOQ) can be used as quality parameters for extracts based on S. chilensis.

Simultaneous determination of quercitrin, phloridzin and 3-hydroxyl phloridzin in leaves of Malus halliana by ultrasound-assisted ionic liquid-reversed phase liquid chromatography / 中国中药杂志

To establish a method for the simultaneous determination of phloridzin, 3-hydroxy phloridzin and quercitrin in leaves of Malus halliana by ultrasonic-assisted ionic liquid coupled with RP-HPLC. An Agilent TC-C₁₈ (4.6 mm×250 mm, 5 μm) column was used, with the mobile phase of acetonitrile and 1% phosphoric acid-water (20∶80) by gradient elution at the detection wavelength of 270 nm. The flow rate was 0.8 mL•min⁻¹, and chromatographic column temperature was controlled at the room temperature. Under the optimized conditions, the linear ranges for phloridzin, 3-hydroxy phloridzin and quercitrin were 0.9-112.5 μg (r = 0.999 6), 0.093 2-11.65 μg (r = 0.999 1) and 0.097 2-12.15 μg (r = 0.999 8), respectively. The average recoveries of the three constituents were 99.35%, 98.80% and 98.19%, respectively. The method was environmental friendly, rapid, accurate and highly reproducible, and so suitable for the quantitative analysis of phloridzin, 3-hydroxy phloridzin and quercitrin in leaves of M. halliana.