Quality evaluation of Huaijiao pill by chromatographic fingerprint and simultaneous determination of its major bioactive components

For quality control purpose,an approach of combining chromatographic fingerprint of Huaijiao pill（HP）and simultaneous determination of its major bioactive components was developed using high performance liquid chromatography coupled with diode array detector（HPLC-DAD）.For fingerprint analysis,16 peaks were selected as the characteristic peaks to evaluate the similarities of different samples collected from different batches of three manufacturers.The similarities of 17 Huaijiao pill samples were beyond 0.966,indicating that samples from different batches and manufacturers were,to some extent,consistent.Additionally,simultaneous quantification of seven bioactive markers,namely sophoricoside,baicalin,naringin,genistein,rutin,quercetin and 5-O-methylvisammioside,in HP was performed to interpret the quality consistency.The validation of the proposed approach was acceptable,with the accuracy of 90.2%-106.9%in recovery test.The intra-day and inter-day precisions of the method were evaluated and the RSD values were less than 2.81%.The results from the quantitative data showed that the contents of six marker compounds（except for 5-O-methylvisammioside） were quite consistent between batches produced by one manufacturer and significantly distinctive among different manufacturers.The proposed approach was expected to be developed as a powerful tool for the quality control of HP.

Evaluation of reservoir connectivity using whole-oil gas chromatographic fingerprint technology: A case study from the Es_3~3 reservoir in the Nanpu Sag, China

In this study, whole-oil gas chromatographic fingerprint analyses were performed on oils from the Es3^3 reservoir in the Liubei area of the Nanpu Sag. The gas chromatographic peaks of cyclic and branched alkanes with relatively high resolution from nCl0 to nC25 were selected to establish a database of whole-oil gas chromatographic peak height ratio fingerprints. Reservoir fluid connectivity was identified by using clustering analysis. This method can reflect the gas chromatography fingerprint information accurately and entirely, and avoid the one-sidedness of the star diagram method which only selects several fixed gas chromatographic peaks.

Study on chromatographic fingerprint of sarcandra glabra (Thunb.) by microwave-assisted extraction coupled to HPLC/DAD

Microwave-assisted extraction(MAE)was used for extraction of effective components of sarcandra glabra(Thunb.),and then chromatographic fingerprint of sarcandra glabra(Thunb.)was studied by high performance liquid chromatography/diode array detector(HPLC/DAD).The conditions of MAE were optimized by an orthogonal experiment,and then the authentication and validation of the chromatographic fingerprint were conducted.Nine peaks were identified as common peaks in the fingerprint chromatograms,and isofraxidin was considered as a reference compound and quantified.Relative standard deviations of retention time and peak area of each component were less than 3% and 8%,respectively.Similarity and difference analysis were conducted by use of PCA and relation coefficient.Twenty batches of sarcandra glabra(Thunb.)samples from two different producing areas could be classified into two different groups in the PCA model.The results showed that MAE-HPLC/DAD method was simple,efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra(Thunb.),which could provide more reliable and precise information for quality evaluation.

Chemical fingerprinting of Su-He-Xiang-Wan and attribution of major characteristic peaks for its quality control by GC-MS

A simple and facile gas chromatography-mass spectrometer （GC-MS） fingerprint of Su-He-Xiang-Wan （SHXW） was developed, the similarity analysis was conducted, and attribution of the major characteristic peaks was identified for SHXW quality control. GC-MS analysis was performed on a QP2010 instrument （Shimadzu, Japan） equipped with a capillary column of RTX-5MS. The column temperature was initiated at 50℃, held for 5 min, increased at the rate of 3 ℃/min to 120 ℃, held for 2 min, and then increased at the rate of 4 ℃/min to 220℃, held for 10 min. Helium carrier gas was used at a constant flow rate of 1.3 mL/min. Mass conditions were ionization voltage, 70 eV; injector temperature, 250℃; ion source temperature, 250 ℃; splitting ratio, 30：1; full scan mode in the 40-500 Da mass ranges with rate of 0.2 s per scan. Attribution of the major characteristic peaks was identified for SHXW by comparing the chemical standards, references of Chinese herbal medicines and the negative controls of prescription samples （NC） of SHXW. With the help of the temperature-programmed retention indices （PTRIs） used together with mass spectra and chemical standards, 25 major characteristic peaks have been identified. Nine volatile medicinal materials were identified in the prescription of SHXW by attributing to the 27 major characteristic peaks. The results demonstrate that the proposed method is a powerful approach to quality control of complex herbal medicines.

烟用香精的GC-MS指纹图谱

在卷烟生产中,烟用香精对卷烟的风格起着决定作用[1],所以稳定的烟用香精质量对卷烟内在品质的控制非常重要.目前,烟用香精的质量控制以物理指标[2]方法为主,对质量的变化反映不显著,给质量控制带来困难.……

草果挥发油的GC-MS指纹图谱

采用GC-MS方法建立草果挥发油指纹图谱,探讨气相色谱-质谱(GC-MS)连用技术在草果挥发油质量控制方面应用的可行性。采用气质连用的方法,对不同产地的草果油的指纹图谱进行比较分析。草果挥发油指纹图谱共检出10个特征峰,特征峰峰面积之和占总峰面积的90％以上;其精密度、稳定性、重复性均符合有关规定。不同产地草果油的指纹图谱中各成分均得到了较好的分离,具有特征性和唯一性,因此本方法可作为草果油质量的控制方法。

百合弱极性成分GC-MS鉴定及其特征性成分TIC指纹图谱构建

对百合典型材料弱极性成分提取溶剂进行优化与GC-MS检测分析,所得总离子色谱图成分峰通过工作站自动检索NIST2008质谱库进行定性分析,采用色谱峰面积归一法测定各成分相对含量,并采用"中药色谱指纹图谱相似度评价系统2004A版"计算全国12个主产区百合弱极性成分石油醚提取物总离子色谱图的整体相似度,构建百合弱极性成分石油醚提取物总离子色谱指纹图谱.结果表明:(1)百合弱极性成分最佳提取溶剂为石油醚;(2)GC-MS检测共鉴定出百合弱极性部位化合物53个,主要成分为棕榈酸(8.68%)、亚麻酸(7.55%)、γ-谷甾醇(4.54%)、柠檬烯(1.66%)、硬脂酸(4.17%)及其同系物;(3)12批百合样品弱极性成分石油醚提取物总离子色谱图成分峰分布均匀,分离度高,分离效果好;主要成分峰均实现共有性匹配,匹配共有成分峰39个,匹配结果良好;各样品色谱图与所构建对照图谱相似度均在0.875以上.

天然沉香香气成分的SHS-GC-MS指纹图谱研究

目的建立基于静态顶空进样-气相色谱-质谱（SHS-GC-MS）联用技术的天然沉香香气成分指纹图谱。方法以22批经《中国药典》及前期研究方法验证为天然沉香的沉香样品为材料,采用SHS-GC-MS联用技术构建天然沉香香气成分指纹图谱,并用《中药指纹图谱相似度评价（2004 A版）》软件进行数据处理;采用自动质谱退卷积定性系统（AMDIS）结合保留指数（RI）鉴定共有峰,采用主成分分析（PCA）对不同产地天然沉香进行判别分析,并将共有峰的相对峰面积与浸出物质量分数进行相关性分析。结果 22批天然沉香指纹图谱的相似度范围为0.467~0.905,经AMDIS结合RI双重定性鉴定了18个色谱峰。以全谱碎片信息和18个共有峰相对峰面积为变量分别进行PCA分析,22批天然沉香按产地均可分为2类。共有成分γ-桉叶油醇与沉香的主要评价指标——浸出物质量分数呈显著正相关。结论18个共有峰组成的香气成分指纹图谱能表达天然沉香的共有品质并区分不同产地的天然沉香,可为沉香的感官分析提供简便、快速的科学依据。γ-桉叶油醇可作为香气指纹图谱的指标性成分。

嗜热链球菌发酵乳中挥发性风味物质的GC-MS指纹图谱

嗜热链球菌是发酵乳生产中常用的发酵剂菌种之一。以分离自传统发酵乳制品曲拉中的1株具有良好风味的嗜热链球菌IMAU80285为试验菌株,采用气相色谱-质谱联用仪等仪器和主成分分析、相似度分析等方法,对该菌株在发酵乳制作和贮藏期间产生的风味物质进行全面、系统的分析。结果表明:在发酵乳制作和贮藏期间,从嗜热链球菌IMAU80285中共测得84种风味物质,主要包括酸类化合物(11种)、醛类化合物(13种)、酮类化合物(12种)、醇类化合物(22种)、酯类化合物(9种)、烷烃类化合物(14种)、含氮类化合物(3种)。主成分分析结果表明:关键性风味物质(OAV≥0.1)在发酵初期(发酵4,6 h)和前发酵与贮藏期间(后熟,贮藏1,2,3,7,14 d)有明显的区分,发酵初期(发酵4,6 h)和前发酵时,与乙偶姻、庚醛、1-壬醇、己酸乙酯等17种关键性风味物质(OAV≥0.1)有较强的相关性,而贮藏期间(后熟,贮藏1,2,3,7,14 d)与2,3-丁二酮、3-甲基-1-丁醇、丁酸乙酯等7种关键性风味物质(OAV≥0.1)的相关性较强;重叠色谱图及相似度分析结果表明,发酵乳中的风味物质在发酵乳制作及贮藏期间变化显著,随着贮藏时间的延长,发酵乳的风味差异明显。

Study on the Aromatic Components of Green Plum Wine by HS-SPME-GC-MS

In order to better blend green plum wine and study aromatic components of green plum wine,a qualitative analysis on aromatic components of soaked base liquor,green plum soaked wine,green plum juice,and fermented wine of green plum juice by Head Space Solid-phase Microextraction( HS-SPME) and Gas Chromatograph Mass Spectrometer( GC-MS) was studied in this paper. Experiment results indicated that14,32,17,and 46 kinds of aromatic components were identified respectively from four samples. Different aromatic components determined the special flavor and taste of green plum wine. Unique aromatic components generated in soaking process include benzaldehyde,1-butanol,2-methyl-,S-(-),benzoic acid ethyl ester,and 5-( hydroxymethyl). Special aromatic components in green plum juice were furfural,phenylethyl alcohol,and benzyl alcohol. The aromatic components in fermented wine of green plum juice mainly included phenylethyl alcohol( 6. 941%,relative content of peak area,same below),1-butanol,3-methyl-( 6. 940%),octanoic acid,ethyl ester( 3. 734%),decanoic acid,ethyl ester( 2. 590%),hexanoic acid,ethyl ester( 2. 479%),ethyl 9-decenoate( 2. 080%),and 5-hydroxymethyl( 1. 756%). This study was expected to provide scientific basis and data reference for quality improvement of green plum wine.

Gas Chromatography-Based Fingerprinting and Chemical Pattern Recognition of Rosa multiflora Volatile Oil

In this paper, gas chromatography-mass spectrometry （GS-MS） was used to build the standard fingerprint of volatile oil from Rosa multiflora Thunb. from 12 different habitats. Fourteen components in the volatile oil were identified as the indicator components ofR. multiflora, of which one was selected as the standard. The GC analysis conditions used for fingerprinting afford a very good separating effect. The similarity of the 12 volatile oils from R. multiflora Thunb. was more than 0.84, and the precision, stability and repeatability of the fingerprints were quite good. It could be concluded that the fingerprints can be used as the standard and as a quality control method for medicinal materials from R. multiflora Thunb..

A novel method for integrating chromatographic fingerprint analytical units of Chinese materia medica:the matching frequency statistical moment method

Objective To facilitate the quality evaluation suitable for the unique characteristics of Chinese materia medica(CMM)by developing and implementing a novel approach known as the matching frequency statistical moment(MFSM)method.Methods This study established the MFSM method.To demonstrate its effectiveness,we applied this novel approach to analyze Danxi Granules(丹膝颗粒,DXG)and its constituent herbal materials.To begin with,the ultra-performance liquid chromatography(UPLC)was applied to obtain the chromatographic fingerprints of DXG and its constituent herbal materi-als.Next,the MFSM was leveraged to compress and integrate them into a new fingerprint with fewer analytical units.Then,we characterized the properties and variability of both the original and integrated fingerprints by calculating total quantum statistical moment(TQSM)parameters,information entropy and information amount,along with their relative standard deviation(RSD).Finally,we compared the TQSM parameters,information entropy and infor-mation amount,and their RSD between the traditional and novel fingerprints to validate the new analytical method.Results The chromatographic peaks of DXG and its 12 raw herbal materials were divided and integrated into peak families by the MFSM method.Before integration,the ranges of the peak number,three TQSM parameters,information entropy and information amount for each peak or peak family of UPLC fingerprints of DXG and its 12 raw herbal materials were 95.07−209.73,9390−183064μv·s,5.928−21.33 min,22.62−106.69 min^(2),4.230−6.539,and 50530−974186μv·s,respectively.After integration,the ranges of these parameters were 10.00−88.00,9390−183064μv·s,5.951−22.02 min,22.27−104.73 min^(2),2.223−5.277,and 38159−807200μv·s,respectively.Correspondingly,the RSD of all the aforementioned pa-rameters before integration were 2.12%−9.15%,6.04%−49.78%,1.15%−23.10%,3.97%−25.79%,1.49%−19.86%,and 6.64%−51.20%,respectively.However,after integration,they changed to 0.00%,6.04%−49.87%,1.73%−23.02%,3.84%−26.85%,1.17%−16.54%,and 6.40%−48.59%,respectively.The results demonstrated that in the newly integrated fingerprint,the analytical units of constituent herbal materials,information entropy and information amount were significantly reduced(P<0.05),while the TQSM parameters remained unchanged(P>0.05).Additionally,the RSD of the TQSM parameters,information entropy,and information amount didn’t show significant difference before and after integration(P>0.05),but the RSD of the number and area of the integrated analytical units significantly decreased(P<0.05).Conclusion The MFSM method could reduce the analytical units of constituent herbal mate-rials while maintain the properties and variability from their original fingerprint.Thus,it could serve as a feasible and reliable tool to reduce difficulties in analyzing multi-compo-nents within CMMs and facilitating the evaluation of their quality.

Estimation of the Postmortem Interval Using Chromatographic Fingerprints of Volatile Organic Compounds from Muscle

Estimation of the postmortem interval(PMI)is a crucial task in the field of forensic pathology and has unfortunately not been properly resolved.In this study,we analyzed volatile organic compounds(VOCs)in rat muscle samples collected at different PMIs and studied the feasibility of muscle VOC fingerprinting as a new method for PMI estimation.In total,110 rats were sacrificed and stored at a constant temperature(25℃).Rat skeletal muscle samples were collected at 0-10‑day postmortem,and then the VOCs were determined using a method of headspace solid‑phase microextraction coupled with gas chromatography‑mass spectrometry.The correlations between the VOCs(species and quantities)and PMIs were carefully analyzed and standard muscle VOC fingerprints at 25℃were established for different PMIs.To further test the accuracy of muscle VOC fingerprinting as a method for PMI estimation,ten additional rats with known PMIs were studied.We identified 15 kinds of VOCs and the number of VOC species increased with the PMI.The total peak areas of the VOCs increased significantly with the postmortem day(adjusted R^(2)=0.96-0.97).The mean error of the VOC fingerprinting for PMI estimation was 0.5 days and the mean relative error was 8.33%.We concluded that muscle VOC fingerprinting combining the use of VOC species and peak areas is accurate and effective and could be used as an alternative approach for PMI estimation in forensic practice.Although the preliminary results are encouraging,further studies in human cadavers under real case conditions are needed.

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.

High-performance liquid chromatographic fingerprint analysis of Oxytropis falcata Bunge and Oxytropis chiliophylla Royle

A simple, sensible and reliable HPLC-DAD fingerprint analysis method for the raw materials of Oxytropisfalcata and Oxytropis chiliophylla, both of which were used as ＂Er-Da-Xia＂ in Tibetan medicines, was developed and then subsequently applied to analyze samples collected from different locations or times. 19 common fingerprint peaks for O. falcata, 24 for O. chiliophylla, and 11 for the two herbs were designated respectively, including 7 identified characteristic peaks existing in both herbs and 1 uniquely presenting in O. chiliophylla. Although there were some slight differences in the chemicals of O. falcata and O. chiliophylla, the main components of both herbs were consistent generally. The results provided scientific basis, at least from the chemical point of view, for the reasonablity of two herbs being used as the same drug in Tibetan medicines and for the necessary of further investigation on their detailed chemical and pharmacological differences.

Chromatographic fingerprint analysis of Bupleuri Radix by HPLC-ELSD

To facilitate the species identification and quality assessment of Chaihu （Bupleuri Radix）, a simple and valid chromatographic fingerprint method was developed. The method uses high-performance liquid chromatography coupled with evaporative light scattering detector （HPLC-ELSD） and the data analysis is assisted by professional analytical software recommended by the State Food and Drug Administration （SFDA）. The results indicate that Nan Chaihu raw materials and Chaihu decoction pieces vary markedly in chemical quality, while Bei Chaihu raw materials are relatively more stable. Furthermore, it is obvious that Nan Chaihu is chemically very different from Bei Chaihu, suggesting that Nan Chaihu may not be suitable for medicinal use. In addition, the obvious differences between the chromatograms of decoction pieces and raw materials, especially the peaks between 30 and 40 rain and after 45 rain, indicate possible effects of the processing procedures on the chemicals. By analyzing the fingerprints of all samples, 12 main saponin-like fingerprint peaks, of which at least three are characteristic peaks of saikosaponins a, c, and d, are proposed to be considered for further characterization and quality evaluation of Chaihu.

离子色谱指纹图谱分析在烟用香精香料品质评价中的应用

为保证烟用香精香料品质的一致性,采用离子色谱法对烟用香精香料样品中的9种有机酸及7种无机阴离子进行分析。1.0 g样品中加入10 mL去离子水,振荡萃取30 min后,水相溶液用0.45μm微孔滤膜过滤,滤液进一步采用RP前处理柱除去基质,经离子色谱仪分离测定。采用Dionex IonPac AS11-HC阴离子交换柱及淋洗液梯度洗脱,9种有机酸和7种常规无机阴离子标准工作溶液的保留时间和峰面积日内精密度(RSD)分别为0.01%~0.69%和1.34%~2.98%,日间RSD分别为0.03%~0.68%和3.54%~5.16%。同时对烟用香精香料标准样品A~D(各5个批次)中有机酸和无机酸的保留时间和峰面积RSD进行考察,得到的保留时间和峰面积日内RSD分别为0.01%~0.71%和2.39%~3.22%,日间RSD分别0.05%~0.81%和3.61%~6.02%。建立了4种烟用香精香料(各5个批次)的指纹谱图库,并采用系统聚类分析和相似度分析法对其他生产厂家样品进行品质评价。结果表明,聚类分析法能有效地从不同生产厂家的产品中筛选出与标准样品质量最相近的样品;实际样品AY3、BY2、CY2及DY1与香精香料标准品性质最接近,品质较好。相似度分析可以对不同生产厂家的产品质量进行更加具体、量化的评价,实际样品AY3、BY2、CY2及DY1与香精香料标准品的化学成分相似度值均大于97.7%,符合相似度评价要求。相较于超声辅助液液萃取-气相色谱法,对于个别特殊样品,离子色谱法能更有效地区分香精香料样品间的品质差异。

灵芝孢子粉和孢子油脂溶性成分分析

分析不同破壁时间、不同来源灵芝(Ganoderma lucidum)孢子粉,灵芝孢子油加工原料(破壁孢子粉)、中间体(制粒后孢子粉、一级萃取孢子油、二级萃取孢子油)和孢子油成品,不同来源灵芝孢子油,不同植物油的脂溶性成分含量和指纹图谱。结果表明:破壁处理能显著增加灵芝孢子粉中脂溶性成分溶出;不同来源灵芝孢子粉中脂溶性成分含量有差异,但不同来源孢子粉指纹图谱相似度均大于0.90,有18个共有峰;灵芝孢子油加工原料(破壁孢子粉)、制粒后孢子粉、一级萃取孢子油、二级萃取孢子油及孢子油成品脂溶性成分指纹图谱有22个共有峰,不考虑孢子油成品中维生素E峰,孢子油加工原料、中间体和孢子油成品间指纹图谱的相似度均大于0.99;不同来源灵芝孢子油指纹图谱峰组成和峰面积有差异,指纹图谱相似度为0.490~0.925,有23个共有峰;植物油的色谱峰与孢子油相比存在较大差异。研究结果为灵芝孢子粉和孢子油的质量控制提供参考。

油库周缘区地下水烃类污染物示踪研究

油库周缘区地下水中烃类溯源分析对于查明地下水中烃类污染物来源与保障油库安全具有重要现实意义。文章采用色谱与色谱-质谱方法对油库内外地下水浮油与水样中烃类特征组分进行分析,筛选出全烃色谱指纹、饱和烃二环倍半萜指纹和芳烃萘菲系列指纹特征比值,结合不同结构化合物的分子指纹特征比值对油库内外地下水烃类污染来源进行解析研究。结果表明,油库内外地下水浮油的色谱指纹特征、二环倍半萜系列与芳烃萘菲系列化合物特征比值都具有明显相似性,呈现同一来源特征;星图对比油库内外浮油及地下水中的烃类,上述特征比值也呈现相似性,揭示油库周缘区地下水中烃类污染物与库内浮油及地下水中烃类具有较强的亲缘性。目前对于地下水中油烃污染来源鉴别还缺乏一套明确的对比方法,二环倍半萜与芳烃萘菲系列化合物可以作为可靠的油源鉴别指标,为研究地下水油烃污染来源提供一种分析技术手段。

通腑泻下颗粒指纹图谱的建立及7种成分的定量分析

建立通腑泻下颗粒(TongFu XieXia Granules,TFXXG)的高效液相色谱(HPLC)指纹图谱,并同时测定大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、厚朴酚以及和厚朴酚的含量。采用Waters Symmetry C18(2.1 mm×150 mm,3.5μm)色谱柱,以0.1%磷酸水溶液(A)和甲醇(B)为流动相梯度洗脱;流速为0.4 mL/min;柱温为30℃;检测波长为254 nm;进样量为10μL。建立10批TFXXG的指纹图谱,并采用“中药色谱指纹图谱相似度评价系统”(2012版)计算相似度;测定大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、厚朴酚以及和厚朴酚的含量。结果表明,10批TFXXG共确定出25个共有峰,相似度均在0.950以上;共指认出12个共有峰,分别为芥子碱硫氰酸盐、咖啡酸、松果菊苷、柚皮苷、新橙皮苷、芦荟大黄素、大黄酸、厚朴酚、和厚朴酚、大黄酚、大黄素以及大黄素甲醚。其中,芦荟大黄素、大黄酸、厚朴酚、和厚朴酚、大黄素、大黄酚以及大黄素甲醚的含量分别为0.1244~0.1304、0.4271~0.4387、3.0479~3.0853、1.2910~1.3039、0.2427~0.2523、0.7718~0.7897和0.5085~0.5249 mg/g。本文所建指纹图谱和定量分析方法重复性好,可操作性强,可用于控制TFXXG的质量。