HPLC fingerprint of Nauclea officinalis stems in different harvesting period

Objective:To establish HPLC fingerprint of Nauclea officinalis stems in different harvesting periods,and analyze the effect of harvest time on the quality of the medicinal materials by combining with chemical pattern recognition.Methods:The analysis was performed on Sun Fire-C18(150 mm×4.6 mm,5μm)column.The mobile phase consisted of acetonitrile-0.1%phosphoric acid solution(gradient elution)at a flow rate of 1.0 mL/min.Results:The HPLC fingerprint of Nauclea officinalis stems was established and 12 common peaks were determined,and 3 chromatographic peaks were identified by comparison with the mixed references.There were some differences in the quality of Nauclea officinalis in different harvesting periods.The OPLS-DA analysis successfully predicted four main markers of quality difference.Conclusion:The established HPLC fingerprint could reflect the composition characteristics of Nauclea officinalis stems in different harvesting period,and the main markers that influence the composition difference of the stems could be used as key indicators for the quality control.

Correlation analysis between the HPLC fingerprints and in vitro antibacterial activity of ethyl acetate extracts from Radix isatidis

Aim To study the correlation between the HPLC fingerprints and in vitro antibacterial activities of EtOAc extracts of Radix isatidis from various sources. Methods Ten batches of Radix isatidis EtOAc extracts were analyzed with HPLC and the fingerprints were established. The influence of EtOAc extracts on the thermogenic curve of growth of Escherchia coli was obtained by microcalorimetry. The chemical differences of EtOAc extracts of Radix isatidis from various sources in the HPLC fingerprints were probed with hierarchical clustering analysis and similarity analysis. The correlation between the HPLC fingerprints and in vitro antibacterial activities was analyzed with multivafiant correlation analysis. Results Close correlation existed between the HPLC fingerprints and in vitro antibacterial activities of EtOAc extracts of Radix isatidis. Conlusion The combination of HPLC fingerprints and antibacterial activities can be used to discover principle components of Radix isatidis on bioactivity.

Nutritional Compositions of Maca Grown in Ebian and HPLC Fingerprints of Macaene and Macamide

In the present study, the nutritional compositions of maca which was grown in a mountain area at an elevation of 2 200-2 800 m of Ebian County,Sichuan Province were measured, and then HPLC analysis on two representative active compounds（macaene and macamide） in the maca sample was performed.The results revealed that there were 24.20% total protein, 18.40% total amino acids（including 3.84% arginine）, 42.80% total sugars, 1.36% fat and kinds of minerals（including 1.14% potassium） in Ebian maca. HPLC fingerprints of macaene and macamide of Ebian maca were similar to those of Peru maca. The results suggested that maca could be cultivated with good quality in some mountain areas with an altitude1 000 m lower than the origin place in Peru.

Authentication of Cassia seeds on the basis of two-wavelength HPLC fingerprinting with the use of chemometrics

High performance liquid chromatographic(HPLC) fingerprints of Cassia seed,a traditional Chinese medicine(TCM),were developed by means of the chromatograms at two wavelengths of 238 and 282 nm.Then,the two data sets were combined into one matrix.The application of principal component analysis(PCA) for this data matrix showed that the samples were clustered into four groups in accordance with the plant sources and preparation procedures.Furthermore,partial least squares(PLS),back propagation artificial neural...

Optimization of high pressure machine decocting process for Dachengqi Tang using HPLC fingerprints combined with the Box–Behnken experimental design

Using Dachengqi Tang（DCQT） as a model, high performance liquid chromatography（HPLC）fingerprints were applied to optimize machine extracting process with the Box–Behnken experimental design. HPLC fingerprints were carried out to investigate the chemical ingredients of DCQT; synthetic weighing method based on analytic hierarchy process（AHP） and criteria importance through intercriteria correlation（CRITIC） was performed to calculate synthetic scores of fingerprints; using the mark ingredients contents and synthetic scores as indicators, the Box–Behnken design was carried out to optimize the process parameters of machine decocting process under high pressure for DCQT. Results of optimal process showed that the herb materials were soaked for 45 min and extracted with 9 folds volume of water in the decocting machine under the temperature of 140 1C till the pressure arrived at 0.25 MPa;then hot decoction was excreted to soak Dahuang and Mangxiao for 5 min. Finally, obtained solutions were mixed, filtrated and packed. It concluded that HPLC fingerprints combined with the Box–Behnken experimental design could be used to optimize extracting process of traditional Chinese medicine（TCM）.

An HPLC Fingerprint Identification of Dried Barks of Ilex rotunda and Ilex godajam

[Objectives] A simple and reliable HPLC fingerprint method was developed for the identification of dried barks of Ilex rotunda and I. godajam. [Methods] Nine batches of dried barks of I. rotunda,and seven batches of dried barks of I. godajam collected from different pharmacies and arboretums in different regions of China were used to establish fingerprints. The software Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine( 2004 A Edition) was used to evaluate the fingerprints. [Results]The fingerprints of dried barks of I. rotunda and I. godajam were established. Methodological study met the technical requirements of fingerprints. The similarities of the fingerprints of dried barks of I. rotunda and I. godajam were all more than 0. 8 and 0. 9 respectively. There were 31 and 28 common peaks in I. rotunda and I. godajam,which could be classified into two clusters by principal component analysis( PCA) and hierarchical cluster analysis. [Conclusions] The feasibility and advantages of used HPLC fingerprints were verified,and the results indicated that the HPLC fingerprint as a characteristic distinguishing method combining similarity evaluation,principal component analysis and hierarchical cluster analysis can be successfully used to identify the authenticity of dried barks of I. rotunda and I. godajam.

HPLC Fingerprint with Multi-components Analysis for Quality Consistency Evaluation of Traditional Chinese Medicine Si-Mo-Tang Oral Liquid Preparation

Si-Mo-Tang（SMT） oral liquid preparation, a traditional Chinese medicine, was prepared from four crude herbal drugs, Fructus Aurantii Submaturus, Radix Aucklandiae, Semen Arecae and Radix Linderae Aggregatae. A combinative method using HPLC fingerprint and quantitative analysis was developed and validated for quality consistency evaluation of SMT. Individual HPLC chromatograms were evaluated against the mean chromatogram generated via a similarity evaluation computer program. Data from chromatographic fingerprints were also processed with principal component analysis（PCA） and hierarchical cluster analysis（HCA）. Additionally, six components （naringin, isonaringin, hesperidin, neohesperidin, norisoboldine and potassium sorbate） in SMT were simultaneously determined to interpret the quality consistency. For fingerprint analysis, 20 peaks were selected as the characteristic peaks to evaluate the similarities of 26 SMT collected from different manufacturers. Among the 20 characteristic peaks, 10 peaks were assigned to be naringin, hesperidin, neohesperidin, isonaringin, neoeriocitrin, tangeretin, nobiletin, norisoboldine, 5-（ethoxymethyl）furan-2-carbaldehyde and potassium sorbate, respectively. The results of similarity analysis, PCA and HCA, indicate that the samples from different manufacturers were consistent with each other in composition. The results from the quantitative data show that the contents of six compounds were significantly different in SMT oral liquid preparations from different manufacturers. The combinative method of chromatographic fingerprint with quantitative analysis developed here offered an efficient way for the quality consistency evaluation of the traditional Chinese medicine SMT.

Establishment of HPLC Fingerprint, Cluster Analysis and Principle Component Analysis of Citri Reticulatae Pericarpium Viride

[Objectives] This study aimed to establish HPLC fingerprint and conduct cluster analysis and principle component analysis for Citri Reticulatae Pericarpium Viride. [Methods] Using the HPLC method, the determination was performed on XSelect~® HSS T3-C_(18) column with mobile phase of acetonitrile-0.5% acetic acid solution(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was 360 nm. The column temperature was 25℃. The sample size was 10 μL. With peak of hesperidin as the reference, HPLC fingerprints of 10 batches of Citri Reticulatae Pericarpium Viride were determined. The similarity of the 10 batches of samples was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition) to determine the common peaks. Cluster analysis and principal component analysis were performed by using SPSS 17.0 statistical software. [Results] The HPLC fingerprints of the 10 batches of medicinal materials had total 11 common peaks, and the similarity was 0.919-1.000, indicating that the chemical composition of the 10 batches of medicinal materials was consistent. There were 11 common components in the 10 batches of medicinal materials, but their contents were different. When the Euclidean distance was 20, the 10 batches of samples were divided into two categories, S4 in the first category, and the others in the second one. When the Euclidean distance was 5, the second category could be further divided into two sub-categories, S1 and S10 in one sub-category, and S2, S3, S5, S6, S7, S8 and S9 in the other one. The principle component analysis showed that cumulative contribution rate of the two main component factors was 92.797%, and the comprehensive score of S7 was the highest with the best quality. [Conclusions] The results of HPLC fingerprinting, cluster analysis and principle component analysis can provide reference for the quality control of Citri Reticulatae Pericarpium Viride.

Study on HPLC Fingerprint and Simultaneous Determination of Three Active Components of Dipsaci Radix Collected in Different Time

[Objectives] To establish methods of HPLC fingerprint and simultaneous determination of the three bioactive components(namely, asperosaponin VI, loganin and sweroside) of Dipsaci Radix, and explore the quality situation of this crude medicine collected in various months in autumn. [Methods] The analysis was performed on Thermo BDS Hypersil C_(18)(4.6 mm×250 mm, 5 μm) column with a mixture of acetonitrile and 0.1% phosphoric acid as mobile phase in gradient mood at a flow rate of 1.0 mL/min, the column temperature was set at 30 ℃, and the detection wavelength was 220 nm. [Results] The analysis methods for HPLC fingerprint and determination of the three components in Dipsaci Radix had been established. The results showed that 12 batches of samples, which were collected in four different places from September to November, possessed the similarities of greater than 0.976. Through the quantitative analysis of asperosaponin VI, loganin and sweroside in Dipsaci Radix, it was found that the quality variation of this herbal medicine and the different collected months of autumn showed a low correlation. [Conclusions] The established methods of HPLC characteristic fingerprint and simultaneous determination of three glycosides provided a new way for quality analysis of Dipsaci Radix. It was preliminarily indicated that collecting this plant medicine in different months of autumn would not affect its quality.

HPLC Fingerprint Chromatogram of Tetracera asiatica Produced in Guangxi

[Objectives] To establish fingerprint chromatogram of Tetracera asiatica and provide basis for controlling and assessing the quality of Tetracera asiatica.[Methods] The high-performance liquid chromatography( HPLC) method was used. Phenomenex C18( 5 μm,4. 6 ×250 mm),acetonitrile-0. 1% phosphoric acid water,gradient elution,flow rate 1 m L/min,detection wavelength 214 nm,and column temperature 25 ℃. The fingerprint chromatogram of 10 batches of samples of Tetracera asiatica was determined for similarity comparison. [Results] It established HPLC fingerprint chromatogram of Tetracera asiatica produced in Guangxi,determined a total of 13 common fingerprint peaks. The similarity of 10 batches of Tetracera asiatica was > 0. 9.[Conclusions] The method is rapid and accurate and can provide references for assessing the quality of Tetracera asiatica.

On the HPLC Fingerprint of Zhuang Medicine Spilanthes paniculata Wall. ex DC.

[Objectives] To establish the HPLC fingerprint of Spilanthes paniculata Wall. ex DC. and provide reference for the quality control of medicinal materials. [Methods]Welch Ultimate XB-C18( 5 μm,250 mm ×4. 6 mm) chromatographic column was adopted,and the mobile phase was acetonitrile-0. 1% phosphoric acid aqueous solution for gradient elution,the detection wavelength was 328 nm,the column temperature was 25 ℃,the similarities of 18 different batches of medicinal materials were evaluated,and the HPLC fingerprint chromatogram of S. paniculata Wall. ex DC. was set up. [Results] The RSDs of precision,repeatability and stability of HPLC fingerprint of 18 batches of S. paniculata Wall. ex DC. medicine materials were all less than 3%;HPLC reference fingerprint of the medicine materials was established.[Conclusions]HPLC fingerprint was established,its data were stable and reliable,and the method was simple and efficient,which could provide reference for the quality evaluation of Zhuang medicine S. paniculata Wall. ex DC.

Preliminary Study on HPLC Fingerprint of Thlaspi arvense L.

[Objectives] To establish Thlaspi arvense L. HPLC fingerprints,to provide reference for quality evaluation Thlaspi arvense L.[Methods]HPLC chromatography was used,HPLC column Acclaim~ 120C_(18)( 250 mm × 3. 0 mm,5 μm),mobile phase of acetonitrile-0. 2% phosphoric acid aqueous solution gradient elution,flow rate of 0. 7 m L/min,column temperature of 30 ℃,detection wavelength of 320nm; the chromatographic fingerprint similarity evaluation software( Version 2004 A) was used for similarity evaluation and data processing on 10 origin Thlaspi arvense L. spectra,and the median method was used to generate control Thlaspi arvense L. fingerprint. [Results] The similarity of 10 origin Thlaspi arvense L. spectra was greater than 0. 90,19 common peaks were separated from different origin Thlaspi arvense L. and isovitexin,apigenin,luteolin and acacetin were identified. [Conclusions] The established HPLC fingerprint contained a lot of information and showed good specificity for Thlaspi arvense L.,which can provide a scientific basis for Thlaspi arvense L. quality evaluation system.

HPLC多波长切换指纹图谱结合化学计量学研究白术土炒前后成分变化规律

目的 建立测定白术、土白术水溶性和脂溶性成分的高效液相色谱-二极管阵列检测器(HPLC-DAD)多波长切换指纹图谱,结合化学计量学研究白术土炒前后成分变化规律。方法 建立白术、土白术水溶性及脂溶性成分的HPLC-DAD多波长切换指纹图谱测定方法,测定17批白术和17批土白术的HPLC指纹图谱,采用相似度评价、聚类分析、OPLS-DA分析进行统计分析。结果 白术指纹图谱中共标定9个共有峰,土白术指纹图谱中共标定10个共有峰。指认了新绿原酸、绿原酸、白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ及苍术酮6个成分,土炒后1号峰、4号峰(白术内酯Ⅲ)、8号峰(白术内酯Ⅱ)峰面积增加,2号峰(新绿原酸)、3号峰(绿原酸)、5号峰、6号峰、7号峰、9号峰(白术内酯Ⅰ)、10号峰(苍术酮)土炒后峰面积均降低。结论 所建立指纹图谱方法能够系统地分析白术土炒前后化学成分的变化规律,可为进一步规范白术土炒工艺,制定土白术专属性质量标准,研究土白术炮制原理提供实验基础。

引火汤冻干粉HPLC指纹图谱建立及8种成分含量测定

目的建立引火汤冻干粉HPLC指纹图谱,并测定地黄苷D、麦冬甲基黄烷酮A、毛蕊花糖苷、原儿茶酸、梓醇、五味子醇甲、五味子乙素、五味子甲素的含量。方法指纹图谱建立采用Supersil AQ18色谱柱(4.6 mm×250 mm,5μm);流动相乙腈-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长230 nm。UPLC-MS/MS含量测定采用Alphasil VC-C_(18)色谱柱(2.1 mm×100 mm,2.5μm);流动相乙腈-0.1%甲酸,梯度洗脱;体积流量0.3 mL/min;柱温30℃;电喷雾离子源;正负离子扫描;多反应监测模式。结果10批冻干粉指纹图谱中有17个共有峰,相似度大于0.90,指认出毛蕊花糖苷、水晶兰苷、去乙酰车叶草苷酸、五味子醇甲。8种成分在各自范围内线性关系良好(R^(2)>0.9906),平均加样回收率98.7%~101.1%,RSD 2.0%~5.2%。结论指纹图谱结合含量测定能完整表征引火汤基准样品质量,从而为其关键化学属性质量评价提供参考。

基于HPLC特征图谱和多成分含量评价梨干的质量

目的建立梨干的特征图谱,测定其中熊果苷、绿原酸、芦丁3种成分的质量分数,结合化学模式识别方法,综合评价其质量差异。方法采用HPLC法,乙腈-0.1%(体积分数)磷酸水溶液作为流动相进行梯度洗脱,流速:1.0 mL/min;柱温:30℃;检测波长:0~25 min,280 nm;26~90 min,350 nm;进样量:20μL。采用“中药色谱特征图谱相似度评价系统(2012版)”软件,结合聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)综合评估20批梨干样本的HPLC特征图谱,同时测定其熊果苷、绿原酸、芦丁的质量分数。结果建立了20批梨干药材的特征图谱,相似度为0.959~0.997,标定了16个共有峰,同时指认了3个共有峰,分别为熊果苷、绿原酸、芦丁,其质量分数分别为16.78~66.00、47.69~260.10、1.39~5.48μg/g,绿原酸质量分数明显高于其他2种成分。通过HCA、PCA和OPLS-DA分析,把20批梨干分为2类(河北产地与非河北产地),并筛选出VIP值大于1的差异性标志成分6种,分别为峰9(绿原酸)、峰4、峰5、峰8、峰13、峰1(熊果苷)。结论本方法简便、准确,可用于梨干的质量控制。

不同产地巴戟天HPLC指纹图谱及化学模式识别研究

目的:利用HPLC指纹图谱及化学模式识别对巴戟天药材进行评价。方法:采用Shodex Asahipak NH2P-504E色谱柱(4.6 mm×250 mm,5μm),以乙腈-水为流动相梯度洗脱,使用蒸发光散射检测器(ELSD)。利用相似度评价、聚类分析和主成分分析等化学模式识别方法,对不同产地的巴戟天药材进行分析。结果:所建立的HPLC指纹图谱共识别了18个共有峰,经与对照品图谱比较,鉴定其中的4个共有峰分别为果糖、蔗糖、蔗果三糖和耐斯糖。6个产地巴戟天的相似度在0.861~0.995;经聚类分析和主成分分析,6个产地巴戟天共聚为3类。结论:本研究采用的巴戟天HPLC指纹图谱及化学模式识别技术可反映出不同产地的巴戟天样品的差异,可为巴戟天的质量控制提供参考。

基于HPLC指纹图谱结合化学模式识别以及多指标成分定量评价前胡药材质量

基于HPLC指纹图谱和多指标成分含量测定,并结合化学模式识别,评价不同产地前胡药材质量。采用Agilent SB-C_(18)(4.6 mm×250 mm,5μm)色谱柱,以甲醇-0.5%甲酸水为流动相进行梯度洗脱,流速为0.5 mL/min,检测波长321 nm,建立指纹图谱并对4个香豆素类成分进行定量,采用聚类分析(hierarchical clustering analysis,HCA)、主成分分析(principal component analysis,PCA)、偏最小二乘法-判别分析(partial least squares discriminant analysis,PLS-DA)对不同产地前胡药材进行质量评价。结果显示16批前胡药材HPLC指纹图谱相似度为0.966~0.999,确定了21个共有峰,共指认了9个成分。通过化学模式识别分析,将16批样品聚类为三类:以安徽宁国、亳州等为主的道地产区前胡,以湖南、贵州、重庆等为主的前胡,以及高海拔云南产区的前胡,并确立8个差异性质量标志物。多指标含量测定不同产地前胡的佛手柑内酯、白花前胡甲素、白花前胡乙素、白花前胡素E含量分别为0.0167~0.1319 mg/g、6.0696~16.2255 mg/g、0.4265~2.0100 mg/g、1.3407~3.1155 mg/g。本研究所建立的指纹图谱和含量测定方法专属性强,稳定性高,分离度良好,结合化学模式可用于前胡药材的质量评价。

小儿热速清口服液HPLC指纹图谱的建立及多指标成分的含量测定

目的建立小儿热速清口服液高效液相色谱(HPLC)指纹图谱,并对其中12种指标成分进行含量测定。方法采用HPLC法。以Venusil MP C18为色谱柱;以乙腈-0.1%磷酸溶液为流动相进行梯度洗脱;流速为1.0 mL/min;检测波长为210 nm;柱温为30℃;进样量为10μL。采用《中药色谱指纹图谱相似度评价系统(2012版)》建立13批小儿热速清口服液的HPLC指纹图谱,并进行相似度评价,测定12种指标成分,即(R,S)告依春、3,5-二-O-咖啡酰奎宁酸、葛根素、连翘苷、连翘酯苷A、绿原酸、黄芩苷、柴胡皂苷d、汉黄芩苷、黄芩素、大黄素、大黄酚的含量。结果13批小儿热速清口服液指纹图谱的相似度均大于0.97,指认了14个共有峰。13批小儿热速清口服液中上述12种指标成分的含量分别为0.078~0.172、1.564~2.736、1.338~2.578、0.426~0.872、1.477~2.628、1.396~2.447、4.052~9.146、0.367~0.692、1.974~4.674、1.274~2.969、0.085~0.167、0.155~0.307 mg/mL。结论本研究建立的HPLC指纹图谱及含量测定方法准确度高、专属性好,可用于小儿热速清口服液的质量评价。

HPLC指纹图谱结合多指标含量测定的抵当芪桂汤质量评价研究

目的 建立抵当芪桂汤HPLC指纹图谱并结合化学模式识别技术对其进行分析与评价,测定抵当芪桂汤中5种有效化学成分的含量,为其质量控制提供依据。方法 采用Agilent 5 TC-C18(2)色谱柱(250 mm×4.6 mm),以乙腈-0.1%磷酸水为流动相进行梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长260 nm。采用SPSS26.0和SIMCA14.1软件对10批抵当芪桂汤进行聚类分析和主成分分析,并通过正交偏最小二乘-判别分析(OPLS-DA)筛选批间差异的标志性成分。结果 10批抵当芪桂汤的HPLC指纹图谱相似度为0.828~0.989,共标定18个共有特征峰,经对照品比对,指认芍药苷、毛蕊异黄酮苷、橙皮苷、桂皮醛、芦荟大黄素5个指标性成分,并对其进行定量分析,线性范围分别为10.000 0~320.000 0μg/mL、2.500 0~80.000 0μg/mL,10.000 0~320.000 0μg/mL、10.000 0~320.000 0μg/mL、0.078 1~5.000 0μg/mL,平均加样回收率为100.30%~104.09%。聚类分析和主成分分析将10批抵挡芪桂汤样品分为2类,通过OPLS-DA筛选出毛蕊异黄酮苷、芍药苷、橙皮苷为质量差异的标志性成分。结论 本研究建立的抵当芪桂汤质量评价方法简便、灵敏、准确、重复性好,可为抵当芪桂汤的质量评价提供依据。

基于HPLC-Q-TOF/MS多成分含量和指纹图谱的加味二妙颗粒质量评价

目的 建立加味二妙颗粒指纹图谱以及多成分含量测定方法,为其质量控制提供参考。方法 采用HPLC-Q-TOF/MS分析加味二妙颗粒中的化学成分。色谱柱为Wondasil C18Superb(250 mm×4.6 mm, 5μm)柱,流动相为乙腈-体积分数0.1%甲酸溶液,梯度洗脱,对10批加味二妙颗粒进行指纹图谱和4种指标性成分含量测定。结果 HPLC-Q-TOF/MS技术共鉴定出加味二妙颗粒中144个化合物,指纹图谱中标定了15个共有峰,并通过对照品比对指认出(R,S)-告依春、去乙酰车叶草苷酸甲酯、盐酸黄柏碱、橙皮苷等9个峰,加味二妙颗粒指纹图谱的相似度均>0.95;测定(R,S)-告依春、落新妇苷、橙皮苷和小檗碱4个指标性成分含量,质量分数分别为0.021 1~0.056 8 mg·g^(-1)、0.200 9~0.552 3 mg·g^(-1)、0.274 1~0.719 1 mg·g^(-1)、0.237 1~0.704 9 mg·g^(-1)。结论 建立的加味二妙颗粒指纹图谱和多成分含量测定方法能够全面反映加味二妙颗粒的质量特征,亦为建立其制剂质量标准提供了依据。