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Iron overload injury is considered to be a part of blood stasis syndrome of arthralgia in traditional Chinese medicine. Its primary therapies include clearing heat and detoxification, activating blood circulation, and removing blood stasis. Lonicera japonica flos (LJF) has long been known as an excellent antipyretic and antidote. Luteoloside (Lut) is one of the main components of LJF and exhibits antioxidant, anti-inflammatory, and cytoprotective properties. However, the protection of Lut against iron overload injury and its underlying mechanisms remain unclear. Therefore, HUVECs were exposed to 50 μmol·L-1 iron dextran for 48 h to establish an iron overload damage model and the effects of Lut were assessed. Our results showed that 20 μmol·L-1 Lut not only increased cell viability and weakened LDH activity, but also significantly up-regulated DDAHⅡ expression and activity, increased p-eNOS/eNOS ratio and NO content, and reduced ADMA content in HUVECs exposed to iron overload. Furthermore, Lut significantly attenuated intracellular/mitochondrial ROS generation, improved SOD, CAT, and GSH-Px activities, reduced MDA content, maintained MMP, inhibited mPTP opening, prevented cyt c from mitochondria released into cytoplasm, reduced cleaved-caspase3 expression, and ultimately decreased cell apoptosis induced by iron overload. The effects of Lut were similar to those of L-arginine (an ADMA competitive substrate), cyclosporin A (a mPTP blocker agent), and edaravone (a free radical scavenger) as positive controls. However, addition of pAD/DDAH II-shRNA adenovirus reversed the above beneficial effects of Lut. In conclusion, Lut can protect HUVECs against iron overload injury via the ROS/ADMA/DDAH II/eNOS/NO pathway. The mitochondria are the target organelles of Lut's protective effects.
Subject(s)
Humans , Endothelium, Vascular , Glucosides , Iron Overload , Luteolin , Reactive Oxygen SpeciesABSTRACT
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.
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Objective: To establish an UHPLC-MS/MS method for rapid and simultaneous determination of the content of 13 components in Ainsliaea fragrans. Methods: The UHPLC-MS/MS method was performed on UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm) with acetonitrile-water (containing 0.1% formic acid and 5 mmol ammonium formate) as mobile phase for gradient elution. Flow rate was 0.25 mL/min and column temperature was 40 ℃. A triple quadrupole mass spectrometer equipped with electrospray ionization source (ESI) was applied in negative ion mode with the following parameters: ion spray, -4 500 V, ion source temperature, 500 ℃, curtain gas, 344.5 kPa, nubulizer (GS1), 344.5 kPa, heater gas (GS2), 206.7 kPa, and multiple reaction monitoring (MRM) was performed for quantitative analysis of these compounds. Results: Under the optimized MS/MS condition, the linearity ranges of protocatechuic acid, caffeic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 1,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, luteolin, luteoloside and apigenin were 1.099-274.800 ng/mL, 1.006-100.600 ng/mL, 1.080-1 080.000 ng/mL, 83.12-8 312.00 ng/mL, 19.92-996.00 ng/mL, 1.076-269.000 ng/mL, 5.555-555.500 ng/mL, 4.420-4 420.000 ng/mL, 44.76-17 904.00 ng/mL, 48.00-9 600.00 ng/mL, 2.108-210.800 ng/mL, 4.136-413.600 ng/mL, 1.070-107.000 ng/mL (r ≥ 0.9958), respectively, with good linearity. The average recovery rate of thirteen compounds in the samples were in the range of 96.54%-99.75%, and the RSD range was from 0.48% to 0.96%. The RSD values of precision, stability and repeatability test were all less than 3.90%. Conclusion: The method had good repeatability, high specificity, stability and controllability, and could be used for quality control of A. fragrans and its preparations.
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Objective: To establish an LCMS/MS method for simultaneous determination of pcoumaric acid, caffeic acid,luteolin and luteoloside in Taraxaci Herba. Methods: The LC analysis was performed on a UPLC-MS/MS instrument with a Dikma Endeavorsil UPLC C18 column(100mm×2.1 mm,1.8 μm),the mobile phase was 0.1% formic acid solution-methanol in a gradient elution,and the flow rate was 0.2 ml/min;the column temperature was set at 35℃; the injection volume was 2 μl. For the MS/MS analysis,the ESI source was operated in the positive-ion mode and the ions were monitored in multiple reaction monitoring(MRM)mode,the temperature of drying gas was 370℃,the sheath gas pressure was set at 10 Bar,the aux gas pressure was maintained at 30 Bar,and the spray voltage was controlled at 4000 V. Results Calibration curves showed good linearity for all 4 compounds in the concentration range of 0.1-10 μg/ml,and the correlation coefficient®was 0.9991,0.9996,0.9994 and 0.9999 for p-coumaric acid,caffeic acid,luteolin,and luteoloside,respectively. Conclusion: The present method could be used for the simultaneous determination of the above mentioned 4 components in Taraxaci Herba.
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Objective: To establish the HPLC specific chromatograms of the ethyl acetate layer in ten batches of effective parts of Filifolium sibiricum and to determine the contents of five components. Methods: The analysis of effective parts of F. sibiricum was performed on a Thermo AcclaimTM120 C18 column (250 mm × 4.6 mm, 5 μm) with acetonitrile (B)-PBS (A) (0.1 mol/L sodium dihydrogen phosphate-2% glacial acetic acid, 1∶1) as the mobile phase in a gradient elution mode, the detection wavelength was set at 360 nm, the flow rate was 0.8 mL/min, and the column temperature was 35 ℃. Results: The specific chromatograms of F. sibiricum effective parts were established and ten common peaks were designated. Among them, five components including isorientin, isovitexin, isoquercitrin, luteoloside and isorhamnetin-3-O-β-D-glucose all showed good linear relationship within the ranges of 0.018—0.108, 0.066 8—0.400 8, 0.088—0.528, 0.118 4—0.710 4 and 0.017 6—0.105 6 μg, respectively. The average recovery was 98.67%, 97.93%, 98.95%, 99.81%, and 97.33% with the RSD value at 1.10%, 0.93%, 1.10%, 0.62%, and 1.48%, respectively. Moreover, the similarity of the eight batches of samples was above 0.9 in the ten batches of medicinal herbs, the similarity of the two batches of which was 0.688 and 0.695, indicating that its content was lower and the difference was greater. In addition, there were significant differences in the content of five components in each harvest time. The content of flavonoids in medicinal herbs was higher with high flower percentage. It was suggested that the content of flavonoids in F. sibiricum was related to the flower percentage of harvest period. Conclusion: The HPLC specific chromatograms of the F. sibiricum effective parts were established and the common characteristic peaks were determined, which could be used for quality control of the F. sibiricum.
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Objective: To optimize the flash extraction process of the active ingredients from Lonicera japonica by Box-Behnken design and response surface methodology.Methods: Flash extraction was used.With the concentration of ethanol,solid-liquid ratio and extraction time as the main influencing factors and the overall normalization value of the transfer rates of chorogenic acid and luteoloside as the evaluation index, Box-behnken design was performed to screen out the optimal extraction conditions.The mathematics relationship between the overall normalization value and the independent variables was established by multiple linear regression and binomial fitting, and response surface methodology was used to predict the optimal process conditions.Results: The optimal extraction conditions were determined as follows: the ethathol concentration was 66.92% ,the solid-liquid ratio was 18.82 ,the extraction duration was 1.20 min and extracted only once.Under the above conditions, the extraction rate of chloragenic acid and total luteoloside was 92.87% and 87.55% , respectively.Conclusion: The optimized technology is simple,economic and practical,which can provide new ideas for the rapid extraction and quality control of active ingredients from Lonicera japonica .
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OBJECTIVE:To establish a method for contents determination of luteoloside,quercetin and hyperoside in Lonicera japonica. METHODS:HPCE was performed silica capillary column with detection wavelength of 360 nm and separation voltage of 20 kV,electrokinetic sampling,sampling voltage of 15 kV,sampling time of 5 s,operation temperature of 25 ℃.The buffer was consisted of 60 mmol/L sodium tetraborate-50 mmol/L natrium carbonicum-50 mmol/L hydroxypropyl-β-cyclodextrin(pH 9.2). RE-SULTS:The linear ranges of luteoloside,quercetin and hyperoside were 0.06-0.56mg/mL (r=0.9881),0.08-0.56 mg/mL (r=0.9892),0.06-0.49 mg/mL(r=0.9796),respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 96.12%-99.77%(RSD=1.29%,n=6),95.90%-98.35%(RSD=0.89%,n=6),94.07%-97.45%(RSD=1.33%,n=6),respectively. CONCLUSIONS:The method is simple,accurate,stable and reproducible,and can be used for simultaneous determination of luteoloside,quercetin and hyperoside in L. japonica.
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OBJECTIVE:To establish a method for simultaneous determination of 5 effective components in Luohua zizhu dry extract. METHODS:UPLC-MS/MS was conducted. The separation was performed on an Zorbax Eclipse Plus C18 column with mo-bile phase of acetonitrile-water(gradient elution)at the flow rate of 0.3 mL/min. The column temperature was set at 40℃and sam-ple size was 2 μL. The analytes were detected in the multiple reaction monitoring(MRM)mode. Nitrogen was used as drying gas and atomized gas. The temperature and flow rate of drying gas were 325 ℃ and 6 L/min. The pressure of atomized gas was 45 psi. The temperature and flow rate of sheath gas were 350℃and 12 L/min. The voltage of capillary were 4000 V(+)and 3500 V(-). The voltage of nozzle was 500 V. RESULTS:The linear ranges of luteoloside,acteoside,quercetin,luteolin and rutin were 0.5048-252.4 ng/mL(r=0.9999),0.7124-356.2 ng/mL(r=0.9990),0.5094-254.7 ng/mL(r=0.9962),0.3030-151.5 ng/mL(r=0.9998) and 0.6022-301.1 ng/mL(r=0.9996),respectively. RSDs of precision,stability and reproducibility tests were all less than 3.0%. The limit of quantitation were 0.42,0.87,0.33,0.12,0.76 ng/mL. The recoveries were 97.99%-101.20%(RSD=1.3%,n=6), 96.50%-101.20%(RSD=1.7%,n=6), 94.81%-99.34%(RSD=1.7%,n=6), 97.54%-100.51%(RSD=1.2%,n=6), 93.37%-98.70%(RSD=1.9%,n=6),respectively. CONCLUSIONS:The method is simple,precise,stable and reproducible, and can be used for simultaneous determination of 5 effective components in Luohua zizhu dry extract.
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Objective: To establish the HPLC chemical fingerprints of Wuwei Xiaoduyin Oral Liquid (WXOL) and simultaneous determination method for six major characteristic components in this herbal preparation, i.e. chlorgenic acid, luteoloside, luteolin, linarin, caffeic acid, and esculetin. Methods: Both chemical fingerprint analysis and quantitative determination were implemented by Agilent 1260 high performance liquid chromatography system with an Agilent 5 TC-C18 column (250 mm × 4.6 mm, 5 μm). A gradient elution program, with the mobile phase of 0.5% glacial acetic acid aqueous solution (A) and methanol (B), was employed as following: 0—10 min, 10%—32% B; 10—20 min, 32% B; 20—25 min, 32%—46% B; 25—31 min, 46%—48% B; 31—41 min, 48%—80% B at the flow rate of 1.0 mL/min. The detection wavelength and column temperature were set at 320 nm and 30 ℃, respectively. Additionally, fingerprint similarity of 10 batches of WXOL was evaluated by software “Similarity Evaluation System for Chromatographic Fingerprint of TCM” published by GPC (Version 2012). The amounts of six characteristic components in WXOL were also simultaneously determined. Results: The common fingerprint pattern derived from 10 batches of samples was obtained with the similarity over 0.98 and 17 common peaks defined. Meanwhile, some common peaks were identified via peak pattern match with standard substances, showing that the peak No.5, No.7, No.8, No.12, No.16, and No.17 was chlorgenic acid, esculetin, caffeic acid, luteoloside, linarin, and lutelin, respectively. Chlorgenic acid content range of 54.038 3—105.551 1 μg/mL, esculetin content range of 4.122 1—31.359 9 μg/mL, caffeic acid content range of 2.413 0—4.420 7 μg/mL, luteoloside content range of 4.042 8—11.312 8 μg/mL, linarin content range of 3.866 3—46.271 9 μg/mL, and luteolin content range of 0.990 8—2.126 8 μg/mL. In combination with the non-significant difference of multiple characteristic components content among 10 batches of samples, the quality of home-made WXOL would be stable. Conclusion: A novel quality control method, which is HPLC fingerprint in combination with simultaneous quantitative analysis of multiple components, was established in this study, with high repeatability and reliability. Therefore, this method provides an applicable approach for the quality control of WXOL.
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AIM To establish an HPLC method for the simultaneous content determination of chlorogenic acid,caffeic acid,luteoloside,baicalein,luteolin and rutin in Yinhuang Granules (Lonicerae japonicae Flos and Scutellariae Radix extracts).METHODS The analysis of 80％ methanol extract of this drug was performed on a 30 ℃ thermostatic SunFire-C1scolumn (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile0.4％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 239 nm.RESULTS All the six constituents showed good linear relationships within their own ranges (r =0.999 9),whose average recoveries were 94.56％-98.38％ with the RSDs of 0.49％-2.89％.Among fourteen batches of samples,the luteoloside content was found to be of little difference,while the other five constituents' contents were of relatively great differences.CONCLUSION The qualities of Yinhuang Granules and Lonicerae japonicae Flos extracts from different manufacturers are uneven,so supervision should be strengthened.
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OBJECTIVE:To establish a method for determining the contents of chlorogenic acid and luteoloside in Lonicerae ja-ponicae,and to explore the shelf life of L. japonicae under ordinary temperature and sealed environment. METHODS:HPLC meth-od was adopted. The determination was performed on Agilent Zorbax SB-C18(chlorogenic acid)column and Agilent Zorbax SB-Phe-nyl(galuteolin)with mobile phase consisted of acetonitrile-0.4% phosphoric acid(13:87,V/V),acetonitrile-0.5% glacial acetic ac-id (gradient elution,galuteolin) at the flow rate of 1.0 mL/min. The detection wavelength was set at 327 nm (chlorogenic acid) and 350 nm(galuteolin). The column temperature was 30 ℃,and sample size was 10 μL. RESULTS:The linear range of chloro-genic acid and galuteolin were 10-100 μg/mL(r=0.9986),5-50 μg/mL(r=0.9993),respectively. RSDs of precision,stability and reproducibility tests were all lower than 4.0%. Recoveries were 95.78%-99.70%(RSD=1.46%,n=6)、96.30%-104.31%(RSD=2.93%,n=6). The contents of chlorogenic acid and galuteolin in roller method,baking method and natural drying method of processed L. japonicae were all decreased by 30%-40% after stored for 12 months. CONCLUSIONS:The method is simple, precise,stable and repeatable,and can be used for simultanoue determination of chlorogenic acid and luteoloside in L. japonicae. The contents of active components in L. japonicae decrease significantly after stored for 12 months. It is necessary to establish shelf life standard of L. japonicae,so as to guarantee the effectiveness of drug use in the clinic.
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Objective: To establish and compare the HPLC characteristic spectra of Humuli Scandentis Herba form different habitats in Hebei province. Methods: HPLC was performed on Agilent 5 TC-C18 (250 mm × 4.6 mm, 5 μm) column, with the mobile phase of acetonitrile -0.2% phosphoric acid at flow rate of 1.0 mL/min; Detection wavelength was 340 nm; The column temperature was 25 ℃ and the sample size was 10 μL. Eleven batches of Humuli Scandentis Herba samples form different habitats in Hebei province were determined and the characteristic spectra of those were established. The fingerprint evaluation software (2004 edition) for Chinese materia medica (CMM) was used to evaluate the similarity of the 11 batches of samples. Results: There were nine characteristic peaks identified in the characteristic spectra of Humuli Scandentis Herba samples. Peak 7 was luteoloside and Peak 8 was apigenin-7-O-β-D-glucosidase. The similarities of seven batches of Humuli Scandentis Herba samples were proved to be higher than 0.900 and four batches of them were proved to be less than 0.900. Conclusion: The method is simple, accurate, and reproducible, and can provide the scientific evidence for controlling the internal quality standards effectively.
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Objective: To establish a method for the determination of luteoloside, apigenin-7-O-β-D-glucosidase, and luteolin by high performance liquid chromatography (HPLC) method, and the determination of luteoloside, apigenin-7-O-β-D- glucosidase, and luteolin in Humulus scandens female and male plants from different origins and different harvest months in Hebei province. Methods: HPLC method was used for simultaneous determination of luteoloside, apigenin-7-O-β-D-glucosidase, and luteolin. System suitability conditions were as follows: column: Agilent 5 TC-C18 (250 mm × 4.6 mm, 5 μm); mobile phase: acetonitrile-0.2% phosphoric acid; flow rate: 1.0 mL/min; detection wavelength: 340 nm; column temperature: 25 ℃; injection volume: 10 μL. Results: The linear ranges were 4.640-74.24 μg/mL of luteoloside (r = 0.999 2), 1.510-30.20 μg/mL of apigenin-7-O-β-D-glucosidase (r = 0.999 9), and 0.796 3-12.74 μg/mL of luteolin. RSD of precision, stability, and repeatabillity tests were ≤ 1.99%. The average recoveries were 102.69% (RSD = 1.64%, n = 9), 100.49% (RSD = 1.93%, n = 9), and 100.39% (RSD = 1.65%, n = 9). Conclusion: The method is simple, reproducible, and accurate, and reliable measurement results can be used for simultaneous determination of luteoloside, apigenin-7-O-β-D- glucosidase, and luteolin in H. scandens female and male plants from different origins and different harvest months in Hebei province. There is greater difference between luteoloside, apigenin-7-O-β-D-glucosidase, and luteolin in H. scandens female and male plants from different origins and different harvest months in Hebei province.
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This study is to establish the HPLC specific chromatogram and determine four main effective components of Lamiophlomis Herba and its counterfeit.Chlorogenic acid, forsythoside B, acteoside and luteoloside were reference substance.HPLC analysis was performed on a Waters XSelect C₁₈ column (4.6 mm×250 mm,5 μm).The mobile phase was acetonitrile-0.5% phosphoric acid solution (18∶82) with isocratic elution.The flow rate was 1.0 mL•min⁻¹, the detection wavelength was 332 nm and the column temperature was 30 ℃.Chemometrics software Chempattern was employed to analyze the research data.HPLC specific chromatogram of Lamiophlomis Herba from different samples were of high similarity, but the similarity of the HPLC specific chromatogram of its counterfeit were less than 0.65.Both of cluster and principal component analysis can distinguish certified products and adulterants.The HPLC specific chromatogram and contents of four effective components can be used for the quality control of Lamiophlomis Herba and its preparations.It provided scientific basis to standardize the use of the crude drug.
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Objective:To study the content determination method for the effective components in WudangⅡFlos lonicerae Caulis to lay foundation for the quality evaluation. Methods: An ultrasonic method was used. The effects of extraction solvent, ultrasonic time, ultrasonic power and ratio of solid to liquid on the contents of rutin and mignonette nucleoside were studied, and the extraction conditions were optimized by a 4-factor and 3-level orthogonal experiment. The chromatographic conditions were as follows:a Phenome-nex Luna-C18(250 mm ×4.60 mm, 5 μm) column was adopted for chlorogenic acid, and a Fortis Xi Phenyl column (250 mm × 4. 6 mm, 5 μm) was used for rutin, loganin and luteoloside;the mobile phase was acetonitrile (B)-0. 4% phosphoric acid (C) solu-tion (15 ∶85) for chlorogenic acid and loganin, and acetonitrile (B) -0. 5% glacial acetic acid aqueous solutjion (D) with gradient e-lution for rutin and luteoloside;the column temperature was 30℃, and the detection wavelength was 327,237,354 and 348 nm, re-spectively. Results:The optimum extraction conditions for rutin and luteoloside from WudangⅡFlos lonicerae Caulis were as follows:the extraction solvent was 60% ethanol, the solid-liquid ratio was 1 ∶30, the ultrasonic power and the ultrasonic time were 350 W and 50 min for rutin, and 250W and 60min for luteoloside. The content of chlorogenic acid, loganin, rutin and luteoloside was 10. 27, 6. 33, 0. 401 and 0. 450 mg·g-1 in the samples, respectively. Conclusion:The method is simple and convenient, accurate and re-producible, which can be used to control the quality of WudangⅡFlos lonicerae Caulis and provide reference for the further develop-ment.
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This study is aimed to explore the mechanism of catalyzing the synthesis of luteolin and luteoloside by LjFNS Ⅱ 1.1 and LjFNS Ⅱ 2.1.The leaves of Lonicera japonica were treated with different concentrations of 5-azaC(20,40,60,80,100 μmol•L-1) for three periods（1,2,3 d）. Firstly, we cloned LjFNS Ⅱ 1.1 and LjFNS Ⅱ 2.1. Secondly, we analyzed the expression levels of LjFNS Ⅱ 1.1 and LjFNS Ⅱ 2.1 by Real-Time PCR and the contents of luteolin and luteoloside determined by UPLC-MS/MS. The results explained the expression levels of LjFNS Ⅱ 1.1 and LjFNS Ⅱ 2.1 consistent with the content variation of luteolin in general, but there was no significant correlation with the contents of luteoloside. And we found the expression levels of LjFNS Ⅱ 1.1 and LjFNS Ⅱ 2.1 were slightly different. The research indicated that the contents of luteolin and luteoloside got higher by improving the expression levels of LjFNS Ⅱ 1.1 and LjFNS Ⅱ 2.1. This will provide technical support and lay a theoretical foundation for regulating the synthesis of luteolin and luteoloside by LjFNS Ⅱ 1.1 and LjFNS Ⅱ 2.1.
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OBJECTIVE:To establish a method for the simultaneous determination of(R,S)-goitrin,chlorogenic acid,luteolo-side and isochlorogenic acid A in Xiaoer ganmao granule. METHODS:HPLC was performed on the column of Hedera C18 with mo-bile phase of acetonitrile-0.1% formic acid aqueous at a flow rate of 1.0 ml/min,the detection wavelength was 254 nm,330 nm and 370 nm,column temperature was 40 ℃,and the injection volume was 5 μl,RESULTS:The linear range was 6.6-105 μg/ml for (R,S)-goitrin (r=0.999 9),9-140 μg/ml for chlorogenic acid (r=0.999 9),9-144 μg/ml for luteoloside (r=0.999 8) and 9-138 μg/ml for isochlorogenic acid A(r=0.999 6);the limits of quantitation were 330 ng,450 ng,450 ng and 450 ng,limits of detection were 66 ng,90 ng,90 ng and 90 ng,respectively;RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 95.01%-98.77%(RSD=1.48%,n=6),95.14%-98.91%(RSD=1.52%,n=6),95.11%-97.54%(RSD=0.93%,n=6) and 95.58%-99.63%(RSD=1.73%,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the simultaneous determination of(R,S)-goitrin,chlorogenic acid,luteoloside and isochlorogenic acid A in Xiaoer ganmao granule.
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Objective: In order to identify the authenticity and control the quality of Xinjiang chrysanthemum, the fingerprinting of it was established using UPLC and discussed in this paper. Methods: Thirteen batches of Xinjiang chrysanthemum from different origins were analyzed by UPLC with catechin, luteoloside, quercetin, and 3, 5-di-O-caffeoyl quinic acid as reference substances. The chromatographic condition was performed on Waters ACQUITY UPLC HSS T3 C18 (150 mm × 2.1 mm, 1.8 μm) and eluted with mobile phase containing acetonitrile-pH 3.0 phosphoric acid solution. The flow rate was 0.2 mL/min with detection wavelength 280 nm in the temperature of 30℃. Results: There were 17 peaks in the UPLC fingerprint of Xinjiang chrysanthemum and three components were identified by three peaks determined using reference substance. Conclusion: The similarity of 13 batches is little difference, but is more than 0.90. The results showed that the UPLC fingerprint chromatogram could be used as a valuable method for the quality evaluation of Xinjiang chrysanthemum.
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Objective To establish a HPLC method for simultaneous determination of chlorogenic acid and luteoloside in the leaves of“Wudang No.II” flos lonicerae. Methods Phenomenex C18(4. 6 mmí250 mm, 5μm) was used;the mobile phase was acetonitrile( A) and 0. 4% phosphoric acid aqueous solution( B) by gradient elution mode; the detection wavelength was 350 nm and the flow rate was 0. 8 mL·min-1;the column temperature was set at 32℃. Results The calibration curve of chlorogenic acid and luteoloside was linear in the range of 0. 285-2. 280μg(r=0. 999 3), and 0. 124-1. 240μg(r=0. 999 4), respectively. The mean recovery of chlorogenic acid and luteoloside was 98. 9%, RSD=1. 59% and 98. 8%, RSD=1. 84%, respectively. Conclusion This method was found to be accurate, quick and reproducible. It can be used for simultaneous determination of chlorogenic acid and luteoloside in the leaves of “Wudang NO.II”flos lonicerae.
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Objective: To develop a UPLC method for the stimultaneous determination of five active ingredients in Callicarpa nudiflora. Methods Analysis was performed on an Agilent Eclipse XDB-C18 column (100 mm ×3.0 mm, 1.8 μm) eluted with acetonitrile (A) and 0.1% methanoic acid (B) in a gradient program. The flow rate was 0.7 mL/min, the detection wavelength was 350 nm, and the column temperature was 40°C. Results Luteoloside, acteoside, luteolin-4'-O-β-D-glucopyranoside, luteolin, and 5, 4'-dihydroxy-3, 7, 3'-trimethoxyflavone showed a good linearity in the ranges of 2.44-122.0 μg/mL (r = 0.999 8), 9.06-453.0 μg/mL (r = 0.999 9), 4.42-221.0 μg/mL (r = 0.999 9), 3.36-168.0 μg/mL (r = 0.999 8), and 2.52-126.0 μg/mL (r = 0.999 8). The average recoveries, measured at three concentration levels, varied from 98.5%-100.8%. Conclusion The method is simple, accurate, and can be used for the quality control of C. nudiflora.