Evaluation and optimization of content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia / 中国中药杂志

This study discovered that the resolution of 3,5-O-dicaffeoylquinic acid(isochlorogenic acid A) in the content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia(ChP)(2020 edition) was poor, which affected accurate quantification. We tested the method in ChP with chromatographic columns of seven brands to clarify the problems in the existing method, optimized the chromatographic conditions by adjusting the mobile phase composition and elution ratio and replacing the chromatographic column packing, and carried out the reproducibility assay for the new method. The two methods were compared for the content determination results of Chrysanthemi Flos prepared from six different varieties. As evaluated by the resolution based on different chromatographic columns of seven brands, the existing method failed to separate isochlorogenic acid A and isochlorogenic acid D well. The peaks of the two components were not completely separated on three chromatographic columns, and isochlorogenic acid A and isochlorogenic acid D generated a co-effluent peak in the other four columns. Isochlorogenic acid A and isochlorogenic acid D could be completely separated under the optimized chromatographic conditions. The difference in the peak areas of isochlorogenic acid A+isochlorogenic acid D obtained by the optimized method and the method in ChP was not significant, with deviation less than 3.0%, which further proved that the result measured by the method in ChP was the co-effluent of isochlorogenic acid A and isochlorogenic acid D. The optimized method can ensure the accurate quantification of isochlorogenic acid A. The existing content determination method of Chrysanthemi Flos has the problem of poor resolution. It is recommended to revise the chromatographic conditions for the content determination method of Chrysanthemi Flos to improve the resolution of isochlorogenic acid A and ensure its accurate quantification.

Isochlorogenic acid (ICGA): natural medicine with potentials in pharmaceutical developments / 中国天然药物

Natural products have attracted a great deal of attention as significant resources in traditional Chinese medicine (TCM) and in chemical medicine, as well as in cosmetic ingredients, nutraceuticals and food products. Isochlorogenic acid (ICGA), which has medicinal value, has been discovered in various plants. As a widespread natural medicine, ICGA should be the subject of further research and development. However, there have been no systematic analyses of ICGA. According to our investigation, ICGA was initially isolated from green coffee extracts by Barnes et al. in 1950. To date, it has been discovered in a variety of tea, vegetables, medicinal diet and TCM materials. ICGA is used as a chemical marker for the quality control of these TCM materials. The metabolic process of ICGA has been studied in detail, conforming to be linear dynamics. Thus, the clear pharmacokinetics of ICGA offers a solid foundation for its research and development. ICGA has multiple biological and pharmacological effects, and studies have mainly focused on its antioxidant, anti-inflammatory, antimicrobial, hypoglycemic, neuroprotective, and cardiovascular protective effects, and hepatoprotective properties. The mechanisms underlying these effects are summarized in this review to provide scientific support and inspiration for the future research and development of ICGA and ICGA-rich natural products.

Simultaneous determination of Forsythoside A and other four components in Xiao'er-Ganmao-Keli from multivendor by HPLC / 国际中医中药杂志

Objective To establish a method for the simultaneous determination of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A by HPLC, and test 16 batches of samples from 14 manufacturers. Methods The test was performed on Kinetex EVO C18 column (150 mm × 4.6 mm, 5 μm) with the column temperature at 35 ℃ . The gradient elution was adopted with the mobile phase of acetonitrile and 0.3% phosphoric acid aqucous at a flow rate of 1.0 ml/min. The detection wavelength of (R,S)-Epigoitrin and Phillyrin were set as 236 nm, the detection wavelength of Forsythoside A, Cholorogenic Acid and Isochlorogenic Acid A were set as 327 nm. Results The good linear relationships were displayed within the linear range of 0.050 45-2.018 00 μg for Forsythoside A (r=0.999 9), 0.018 21-0.728 40 μg for Phillyrin (r=0.999 9), 0.010 16-0.406 40 μg for (R,S)-Epigoitrin (r=0.999 9), 0.006 60-0.263 90 μg for Cholorogenic Acid (r=0.999 9) and 0.0040 44~0.161 76 μg for Isochlorogenic Acid A ( r=0.999 5). The RSDs of reproducibility and stability tests were lower than 2%; recoveries were 97.01%, 98.28%, 99.35% and 96.21%, RSD were 3.19%, 1.19%, 0.81%, 2.88% and 2.96%. The content ranges of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A from 16 batches of samples from 14 manufacturers were 0.057 43-1.508 71 mg/g, 0.017 72-0.350 15 mg/g, 0.005 68-0.177 13 mg/g, 0.007 53-0.226 33 mg/g and 0.00308-0.11908 mg/g. Conclusions The established method is simple and accurate, and has a good repeatability. It can be used for the quality analysis of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A. The content of the tested chemical components from 16 batches of samples from 14 manufacturers have significant differences which indicate that a reinforcement of the quality control is needed.

Isochlorogenic acid A inhibits activation of NLRP3/NF-ΚB in rats with collagen-induced arthritis / 中国药理学通报

Aim To analyse the anti-inflammatory effect of isochlorogenic acid A in rats with collagen-induced arthritis and the potential mechanism . Methods Male SD rats were randomly divided into normal group, model group, isochlorogenic acid A high and low dose groups. The model was constructed in all groups except control group. After continuous administration for six weeks, the plantar thickness and spleen organ coefficient were measured in rats. NLRP3, caspase-1, NF-ΚB p65, P-NF-ΚB p65, P-IΚB and RANKL protein in synovium of joint were detected by Western blot, and IL-lβ, IL-6, TNF-α, CRP, IFN-γ, IL-18 expressions in blood plasma were detected by ELISA kit. Results Compared with normal group, the swelling degree of plantar and the spleen organ coefficient in model group increased significandy. Western blot showed that the expression of NLRP3, caspase-1, NF-ΚB p65, P-NF-ΚB p65, P-IΚB-O and RANKL protein were up-regulated in synovium of joint tissues in model group. ELISA showed that the expression of blood plasma IL-lβ, IL-6, TNF-α, CRP, IFN-γ and IL-18 were significantly up-regulated in model group. Different doses of isochlorogenic acid A can significantly reduce the performance of the above indicators in model group. Conclusions Isochlorogenic acid A has a good anti-inflammatory effect on collagen-induced arthritis, and its anti-inflammatory activity may be related to the decrease of NLRP3 inflammatory complex activation and NF-ΚB phosphorylation.

Simultaneous determination of 12 components in Qiju Dihuang Oral Liquid by HPLC / 中草药

Objective: To establish a rapid, accurate, and practical HPLC method for simultaneous determination the content in Qiju Dihuang Oral Liquid (QDOL) of 5-HMF, morroniside, chlorogenic acid, cryptochlorogenic acid, loganin, paeoniflorin, verbascoside, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and paeonol. Methods: YMC ODS column (250 mm × 4.6 mm, 5 μm) was used, column temperature was set at 35 ℃, gradient elution with 0.1% formic acid aqueous solution- acetonitrile was used as mobile phase, flow rate was 1.0 mL/min, detection wavelength was 254 and 325 nm. The injection volume was 10 μL. Results: The injection amount of 5-HMF, morroniside, chlorogenic acid, cryptochlorogenic acid, loganin, paeoniflorin, verbascoside, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, paeonol injection quality at 0.08—1.60, 0.12—2.40, 0.09—1.80, 0.06—1.20, 0.10—2.00, 0.30—6.00, 0.01—0.20, 0.01—0.20, 0.01—0.20, 0.005—0.10, 0.005—0.10, and 0.01—0.20 μg showed a good linear relationship with peak area, with good precision, repeatability and stability. The recovery rates of the samples were between 96% and 103%, the RSD was 2.13%, 3.45%, 2.86%, 2.59%, 3.15%, 3.49%, 2.19%, 3.25%, 2.37%, 2.53%, 2.91%, and 3.35%, respectively. The content of each component of the five batches of samples was stable, and the mass concentrations range of the 12 components tested were 98.56—102.56, 204.28—212.10, 18.53—18.89, 1.95—2.05, 12.31—12.54, 87.01—87.12, 5.35—5.43, 16.08—16.15, 8.69—8.72, 8.89—8.95, 5.12—5.19, and 1.87—1.94 μg/mL. Conclusion: The method simltaneosly determines the content of 5-HMF, morroniside, chlorogenic acid, cryptochlorogenic acid, loganin, paeoniflorin, verbascoside, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and paeonol in QDOL, which is suitable for the quality control of QDOL.

Simultaneous Determination of Seven Constituents in Qinggan Lidan Mixture by HPLC / 中国实验方剂学杂志

Objective: To establish an HPLC method for simultaneous determination of geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C in Qinggan Lidan mixture,in order to provide references for its quality control. Method: The analysis of methanol extract of this drug was performed on a 35℃ Luna C18 column (4.6 mm×250 mm,5μm),with the mobile phase comprised of acetonitrile-0.4% phosphoric acid flowing at 1.0 mL·min-1 in a gradient elution mode (0-10 min,8%-12%A;10-30 min,12%A;30-60 min,12%-35%A),and the detection wavelengths were set at 238 and 327 nm. Result:Geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C were completely separated,and well separated from other constituents. The linear ranges of geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C were 0.188-2.355,0.083-1.040,0.074-0.920,0.075-0.940,0.064-0.800,0.076-0.955,0.071-0.888 μg (r ≥ 0.999 0),respectively. The average recoveries were 99.45%,98.45%,99.06%,98.50%,98.16%,101.01%,96.93%,with the RSDs of 0.5%,1.8%,1.3%,2.4%,2.3%,1.6%,1.6%,respectively.The contents of geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C were 3.420-3.794,0.835-0.890,1.222-1.275,1.064-1.210,0.377-0.398,0.419-0.464 and 0.362-0.405 g·L-1,respectively. Conclusion:This method can be used for simultaneous determination of muti-ingredients in Qinggan Lidan mixture,and the established method is simple and accurate,with a good reproducibility and high sensitivity. It can be used for the quality control of Qinggan Lidan mixture.

Selection of quality markers of Jasminum amplexicaule based on its anti-diarrheal and anti-inflammatory activities: Effect-target affiliation-traceability-pharmacokinetics strategy / 中草药·英文版

Objective: To investigate therapeutic mechanism in Jasminum amplexicaule (Oleaceae) and verify its main active component as quality control markers Methods: Established mouse models of diarrhea, intestinal angina, and inflammation were firstly used to select herb fractions with optimum efficacy, followed by an in vitro experiment to determine key targets associated with effects of J. amplexicaule extract. The selected fractions were isolated and purified, its components were identified, and the obtained compounds were verified for their effects on NF-κB and iNOS. Finally, effective compounds were administered to rats, their plasma pharmacokinetic parameters were calculated, and quality markers (QMs) reflecting therapeutic activities of J. amplexicaule were confirmed. Results: Trichloromethane and ethyl acetate fractions had significant anti-diarrheal, anti-inflammatory, and analgesic effects. The trichloromethane fraction also reduced BDNF, p38 MAPK, p-p38 MAPK, NF-κB p65, and p-NF-κB p65 levels in the ileum in a rhubarb-induced diarrhea mouse model. Additionally, it inhibited LPS-induced NF-κB transcription and nitric oxide (NO) production in RAW264.7 macrophages, which suppressed iNOS expression. Therefore, the trichloromethane fraction was further investigated. QMs candidate selection identified 17 compounds, and results of in-vitro therapeutic validation indicated that methyl caffeate and isochlorogenic acid B had the strongest anti-diarrheal, anti-inflammatory, and analgesic activities. After being validated by a UHPLC–MS-MS method, concentrations of these target compounds were accurately determined in the rat plasma and pharmacokinetic parameters were calculated. Cmax, tmax, and t1/2 were respectively 575.35 ng/mL (2.963 nmol/mL), 0.5 h, and 0.45 h for methyl caffeate and 262.03 ng/mL (0.5034 nmol/mL), 0.25 h, and 2.03 h for isochlorogenic acid B. Because these candidate compounds exhibited favorable pharmacokinetics, they were considered as QMs of J. amplexicaule. Conclusions: The present study accurately and effectively identified QMs of J. amplexicaule that act as indicators of efficacy and quality.

Rapid determination of seven components in Angelica sinensis based on near infrared spectroscopy / 中草药

Objective: To established a rapid nondestructive determination method for the multi-marker constituents in Angelica sinensis by near infrared spectroscopy (NIRS) combined with the partial least squares (PLS) method in order to improve the quality control for A. sinensis. Methods: A total of 108 batches of A. sinensis from different origins were collected for the research. An Ultra performance liquid chromatography (UPLC) method was established to measure the content of the seven components in A. sinensis, which were adopted as the reference value. And the integrating sphere diffuse reflection mode was employed to collect the NIR spectrum. The quantitative calibration model between the near infrared spectrum and the reference content of each component to be measured was established by PLS and other chemometrics methods. Each part of the modeling process was optimized respectively, including the selection of calibration set and validation set, different pretreatment methods and different spectral section. Results: The correlation coefficient for calibration set of chlorogenic acid, ferulic acid, isochlorogenic acid A, ligustilide, butylidenephthalide, senkyunolide I and levistilide A were 0.937 6, 0.970 2, 0.963 4, 0.991 1, 0.962 4, 0.966 6 and 0.947 6, respectively; The root mean square error of prediction (RMSEP) were 0.072 1, 0.038 9, 0.011 3, 0.483 0, 0.017 5, 0.178 0 and 0.097 0, respectively. The predicted values of NIRS models and the measured values of UPLC showed a good linear relation, which presented a great prediction ability of the models. Conclusion: The methods of NIRS combined with PLS can be applied for the rapid content determination of seven components in A. sinensis including chlorogenic acid, ferulic acid, isochlorogenic acid A, ligustilide, butylidenephthalide, senkyunolide I and levistilide A, which is proved to be simple and reliable.

Simultaneous determination of ten components in Lonicerae Japonicae Flos from different habitats by UPLC / 中草药

Objective: To establish a method for the determination of seven organic acids, one flavone and two iridoid terpenoids in Lonicerae Japonicae Flos from different habitats. Methods: The content of 10 components in Lonicerae Japonicae Flos from different habitats were determined by ultra-performance liquid chromatography (UPLC). The mobile phase for gradient elution was 0.1% phosphoric acid solution (A)- methanol (B); The flow rate was 0.3 mL/min, and the column temperature was 30 ℃. Results: A UPLC method for simultaneous determination of seven organic acids, one flavone and two iridoid terpenoids in Lonicerae Japonicae Flos was established. Principal component analysis (PCA) and partial least squares method (PLS-DA) were used to analyze the distribution and characteristics of 10 constituents of Lonicerae Japonicae Flos collected from different habitats; Three production areas of Shandong, Jiangsu and Shaanxi are respectively grouped into one group. Conclusion: The method is stable and feasible, which could be used as a reference for evaluating the quality of Lonicerae Japonicae Flos in a more comprehensive way.

Determination of Isochlorogenic Acid A and Isochlorogenic Acid C in Vernonia Anthelmintica by High Performance Liquid Chromatography / 医药导报

Objective To establish the chromatographic conditions of isochlorogenic acid A and isochlorogenic acid C in vernonia anthelmintica. Methods By changing the mobile phase,flow rate,column temperature and other chromatographic conditions,the best chromatographic conditions was we pursued to established. Results The linear relationship between the concentration of isochlorogenic acid A and the peak area was between 5. 825-69. 9 μg·mL-1, and the concentration of isochlorogenic acid C,was between 5.15-61.80 μg·mL-1and the peak area was good . The sample recovery rates of the two groups were 98.70%-101.92%(RSD＝1.04%,n＝9)、95.99%-102.52%(RSD＝1.90%,n＝9). Conclusion The method is simple,rapid, accurate and reliable for the determination of isochlorogenic acid A and isochlorogenic acid C in Vernonia anthelmintica and also for the quality control of the raw material.

Accumulation rules of effective constituents in Farfarae Flos of different growth stage based on HPLC characteristic fingerprint / 中草药

Objective To analyze the accumulation patterns of active constituents in Farfarae Flos with different flower bud colors (yellow, purple, and deep purple) at different growth stages, and provide theoretical guidance for the production and quality control of Farfarae Flos. Methods The medicinal materials of Farfarae Flos of different growth stages with different colors were determinated by HPLC method. The Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012 A edition) was used to evaluate the similarity of the samples. The differences among samples were identified by chemical pattern recognition methods including hierarchical principal component analysis (PCA) and partial least squares discriminate analysis (PLS-DA). Results The HPLC fingerprint of different flower bud colors of Farfarae Flos at different growth stages was obtained, 27 common peaks were found in the chromatography, and 11 of them were identified. Similarities of samples of all batches with reference fingerprint were among 0.901-0.995. There were differences in the accumulation characteristics of Farfarae Flos at different growth stages according to the peak area, and showing significant differences among different flower bud colors. PCA and PLS-DA results demonstrated obvious distinction among different flower bud colors. Twelve constituents, such as gallic acid, chlorogenic acid, rutin, hyperin, isochlorogenic acid B and quercetin, were screened as biomarkers, representing major differences among colors. The quality evaluation demonstrated that deep purple buds was the best, followed by purple buds and yellow buds for the worst. Conclusion The HPLC fingerprint can reflect the accumulation characteristics of the active constituents of Farfarae Flos in different growth stages and the differences among different flower bud colors. Combining chemical pattern recognition can provide reference for the production and quality evaluation of Farfarae Flos.

Establishment of UPLC fingerprints of Jasminum elongatum and determination of two constituents / 中成药

AIM To establish the UPLC fingerprints of Jasminum elongatum (Bergium) Wild.and to determine the contents of isochlorogenic acid B and ethylcaffeate.METHODS The analysis of methanol extract of J.elongatum was developed on a 25 ℃ thermostatic Agilent Ecplise XDB-C18 column (2.1 mm × 100 mm,1.7 μm),with the mobile phase comprising of acetonitrile-0.1％ methanoic acid flowing at 0.5 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS There were eighteen common peaks in the fingerprints of ten batches of samples,with the similarities of more than 0.85.Isochlorogenic acid B and ethylcaffeate showed good linear relationships within the ranges of 7.67-38.35 μg/mL (R2 =0.999 4),9.60-96.0 μg/mL (R2 =0.999 7),whose average recoveries were 98.61％,99.09％ with the RSDs of 0.84％,1.25％,respectively.CONCLUSION This stable and reliable method can be used for the quality control of J.elongatum.

Simultaneous Determination of 7 Components in Tanreqing Capsules by HPLC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of 7 components in Tanreqing capsules.METHODS:HPLC method was adopted.The determination was performed on Ultimate XB C18 column with mobile phase consisted of acetonitrile-0.2％ formic acid (gradient elution) at a flow rate of 0.8 mL/min.The detection wavelength was set at 325 nm,and column temperature was 35 ℃.The sample size was 10 μL.RESULTS:The linear ranges of chlorogenic acid,isoforsythiaside A,forsythiaside A,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C and baicalin were 0.025-0.5 μg(r=0.999 6),0.025-0.5 μg (r=0.999 7),0.050-1.0 μg (r=0.999 9),0.025-0.5 μg((r=0.999 7),0.025-0.5 μg ((r=0.999 6),0.025-0.5 μg (r=0.999 6),0.750-1.5 μg(r=0.999 9),respectively.The limit of quantitation was no more than 1.5 ng,and the limit of detection was 0.5 ng.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.The recoveries were 95.28％-106.30％ (RSD ranged 0.97％-2.14％,n=9).CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of 7 components in Tanreqing capsules.

Simultaneous determination of 10 components in Yinhuang preparations by UPLC / 中草药

Objective: To develop an UPLC method for simultaneous determination the contents of 10 active constituents [neochlorogenic acid (NCA), chlorogenic acid (CA), cryptochlorogenic acid (CCA), isochlorogenic acid A (ICAA), isochlorogenic acid B (ICAB), isochlorogenic acid C (ICAC); bacicalin, wogonoside, baicalein, and wogonin] in Yinhuang preparations. Methods: Analysis was performed on an Acquiyt UPLC BEH C18 chromatographic column (100 mm × 2.1 mm, 1.7 μm) eluted with acetonitrile- 0.4% phosphoric acid, gradient elution, and the flow rate was 0.4 mL/min, the detection wavelength was set at 326 nm, the column temperature was 40℃, and the injection volume was 1.0 μL. Results: All calibration curves were linear (r ≥ 0.999 6) over the tested ranges. The average recoveries (n = 9) ranged from 97.43% to 99.94% with RSD value below 2.0%. The contents of 11 batches of NCA, CA, CCA, ICAA, ICAB, ICAC, bacicalin, wogonoside, baicalein, and wogonin were 0.236-3.419, 5.279-26.220, 0.495-4.714, 0.130-2.702, 0.310-3.210, 0.363-5.036, 35.209-133.289, 1.493-6.635, 1.546-5.393, and 0.254-0.823 mg/g. Conclusion: This method is rapid, simple, sensitive, accurate, and reproducible, which can be used for quality control of Yinhuang preparation, and provide a reference for the formulation of quality standards to enhance in the future.

Protective effect of isochlorogenic acid B on CCl4 induced acute liver injury in mice / 国际药学研究杂志

Objective To investigate the effect of isochlorogenic acid B(ICAB)on CCl4-induced acute liver injury(ALI)in mice. Methods The animal model of CCl4-induced ALI in mice was established and then the protective effect of ICAB was evaluated using this model. Serum levels of alanine transaminase(ALT)and aspartate aminotransferase(AST),hepatic levels of malondialdehyde (MDA)and the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were measured by spectrophotometric methods. Liver cell morphology was observed by hematoxylin and eosin staining method and the effects of ICAB on the protein expres-sion of nuclear factor erythroid-2-related factor 2(Nrf2),heme oxygenase-1(HO-1)and quinone oxidoreductase 1(NQO1)in mice he-patocyte were determined by Western blot method. Results ICAB(5,10 and 20 mg/kg)significantly protected against CCl4-induced liver injury by reducing the elevated levels of serum aminotransferases and hepatic MDA and remarkably restored the impaired antioxi-dants. Meanwhile,the histopathological changes were also attenuated in mice. In addition,ICAB could induce the protein expression of Nrf2 and promote its nuclear translocation,and further increase the protein expression of HO-1 and NQO1. Conclusion ICAB has protective effect against CCl4-induced ALI in mice,which is mainly due to its ability to promote the nuclear translocation of Nrf2 and decrease the oxidative stress.

Simultaneous determination of chlorogenic acid, aesculetin, isochlorogenic acid B, and isochlorogenic acid A in Cichorii Herba by QAMS / 中草药

Objective: To establish the quantitative analysis of multi-components by single marker (QAMS) method to determine the content of chlorogenic acid, aesculetin, isochlorogenic acid B, and isochlorogenic acid A in Cichorii Herba (chicory). Methods: Selecting aesculetin as the internal reference substance, the relative correction factors (RCF) of chlorogenic acid, isochlorogenic acid B and isochlorogenic acid A were established respectively, and then the contents of the three constituents were calculated by RCF, to achieve the quality of chicory through QAMS. At the same time, the external standard method was used to determine the content of four constituents in chicory and compare the difference between calculated values and measured values, so as to verify the construction method for accuracy, applicability, and repeatability. Results: No significant difference was observed in chlorogenic acid, isochlorogenic acid B, and isochlorogenic acid A from eight batches of chicory in the quantitative results by these two methods. The validated QAMS method had good precision, reproducibility, and reliability. Conclusion: The established QAMS method is suitable and feasible for the quality control of chicory.

Chemical constituents in stems of Ilex cornuta / 中草药

Objective: To investigate the chemical constituents in the stems of Ilex cornuta and the ability of scavenging free radicals of compounds 1-9. Methods: The compounds were isolated and purified by silica gel, medium pressure column chromatography, and semi-preparative liquid chromatography, and their structures were elucidated by chemical properties and spectroscopic analyses. The antifreeradical efficiency of compounds 1-9 was evaluated by DPPH radical scavenging assay. Results: Fifteen compounds were isolated and their structures were identified as isochlorogenic acid B (1), 3,4,5-tricaffeoylquinic acid (2), 4,5-di-O-caffeoyl quinic acid methyl ester (3), 3,4-di-O-caffeoyl quinicacid methyl ester (4), 3,5-di-O-caffeoyl quinicacid methyl ester (5), 3,4,5-tri-O-caffeoyl quinic acid methyl ester (6), 3,5-dimethoxy-4-hydroxybenzaldehyde (7), ethyl gallate (8), dihydrosyringenin (9), 2,6-dimethoxy-1,4-benzoquinone (10), arctigenin (11), 1-O-(vanillic acid)-6-O-(3″, 5″-dimethoxy-galloyl)-β-D-glycoside (12), (+)-1-hydroxypinoresinol-1-O-β-D-glucopyranoside (13), (+)-(7S,8S)-syringylglycerol 8-O-β-D-glucopyranoside (14), and schaftoside (15). Compounds 1-7 had good antifreeradical efficiency. Conclusion: Compounds 6,8-10,14, and 15 are obtained from the plants of Ilex L. the first time, and compounds 2,7,11, and 12 are obtained from this plant for the first time. Compounds 1-6 have good antifreeradical efficiency.

Analysis on chemical constituents in Bupleuri Radix by HPLC-Q-TOF-MS / 中草药

Objective: To investigate the chemical constituents in extract of Bupleuri Radix using HPLC-Q-TOF-MS. Methods: The HPLC separation achieved on Agilent 1200 HPLC Diamonsil II C18 column (250 mm×4.6 mm, 5 μm) was used with a mobile phase of 0.05% H2O-formic acid (A) and acetonitrile (B), the flow rate was 1.0 mL/min, the column temperature was 35℃, and the detection wavelength was 200-600 nm. Positive and negative ions MS information and element analysis, via comparing MS data with those of the standard compounds and coupled with the related literature were used to analyze the main chemical constituents of BupleuriRadix. Results: Twenty-one chemical constituents were identified, including three phenolic acid, four flavonoids, and fourteen triterpenoid saponins, and isochlorogenic acid A (5), isochlorogenic acid B (6), 7,3'-di-O-methylquercetin (8), and 5-hydroxy-7-acetoxyflavone (24) were separated from Bupleuri Radix for the first time. Conclusion: It is used to quickly analyze the chemical constituents from Bupleuri Radix by establishing a rapid accurate evaluation method using HPLC-Q-TOF-MS, which could provide the references for the application development.

Quality assessment of Shuangyu Granules based on simultaneous determination by HPLC fingerprint and multi-components / 中草药

Objective: To establish an HPLC fingerprint chromatography and determine seven compounds of multi-components in Shuangyu Granules (SG). Methods: The Kromasil C18 (250 mm × 4.6 mm, 3.5 μm) column was used with a mobile phase of acetonitrile-0.05% trifluoroacetic acid gradient elution. The flow rate was 0.8 mL/min, the column temperature was 30℃, and the detection wavelengths were 230 and 327 nm. The common peaks were identified by Q-TOF/MS. Results: The fingerprint chromatography included 17 mutual peaks, and the similarity was more than 0.95. Fourteen common peaks had been identified by LC-Q-TOF/MS, seven of which were unequivocally identified via comparing the retention times and mass spectra data with those of the standard compounds. Then the seven marker components were quantified. The developed quantitative method was validated in terms of accuracy (the recoveries ranged from 97.8% to 101.8% with RSDs less than 2%). Conclusion: The method is rapid, simple, and accurate and can be used for the quality control of SG.

Simultaneous Determination of 4 Components in Xiaoer Ganmao Granule by HPLC / 中国药房

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.