双定性双定量相似度法评价银杏达莫注射液的高效液相色谱指纹图谱

建立了色谱指纹图谱的双定性双定量相似度评价法,并应用于银杏达莫注射液的高效液相色谱（HPLC）指纹图谱的评价。采用反相HPLC,以芦丁为参照物峰,确定了41个共有峰,建立了银杏达莫注射液的对照指纹图谱。以双定性相似度S和S′、双定量相似度C和P评价银杏达莫注射液的HPLC指纹图谱,分别考察在大峰缺失和小峰缺失两种情况下,4个相似度指标的变化特征。S能反映化学成分的分布比例,受大峰影响严重,无法反映小峰的丢失; S′对所有指纹峰等权,反映小峰缺失灵敏,二者构成双定性相似度。C能反映样品共有峰的总体含量,但受大峰影响严重,无法反映小峰的缺失; P对所有峰积分值等权,能较好地反映小峰的变动,二者构成双定量相似度。因此,由S与S′、C与P构成的双定性双定量相似度法能同时监测大峰和小峰的变动与缺失,能准确地解决色谱指纹图谱的宏观定性和定量评价问题。同时还提出了方向余弦作为对照指纹图谱的特征指纹的概念和分解相似度的概念,以此考察了各指纹峰对相似度贡献的大小及其在不同程度缺失时4种相似度的变化情况。所建立的HPLC对照指纹图谱可用于银杏达莫注射液的质量控制。     

高效液相色谱-线性指纹定量法评价人参归脾丸的三波长指纹图谱

以样品指纹向量与对照指纹向量的多元化学组分间存在线性函数关系为基础,引入相关系数r、斜率a、截距b及线性定性相似度S_L、线性定量相似度M_L等概念,建立线性指纹定量法(linearly quantified fingerprint method, LQFM)模型以定性、定量控制中药质量。用高效液相色谱-二极管阵列检测法(HPLC-DAD)同时测定了10批人参归脾丸(Renshen Guipi Wan, RSGPW)分别在203、228和326 nm下的高效液相色谱指纹图谱,运用LQFM理论模型,鉴定RSGPW质量。LQFM能显著区别不同来源的产品,同一厂家产品质量控制一致。鉴定结果与已知的系统指纹定量法评价结果对比,LQFM对中药多元化学成分的复杂系统能实现客观的定性、定量评价。多波长指纹图谱能最大限度地兼顾中药各成分的紫外吸收特征,对中药质量控制具有更全面和可靠的特点。     

清开灵注射液HPLC/ELSD指纹图谱建立及质量相关性研究

运用高效液相色谱／蒸发光散射检测器(HPLC／ELSD)建立了复方清开灵注射液的指纹图谱。对其中7种有效成分进行了定量测定,方法学考察结果良好;通过指纹图谱相似度评价软件,对10批清开灵注射液的HPLC／ELSD指纹图谱进行了相似度计算,结果表明：10批样品相似度很好。该方法为中药质量控制提供了一种可以同时实现整体定性、指标成分定量且简便易行的方法模式。     

基于信息融合的中药多元色谱指纹图谱相似性计算方法

摘要用信息融合算法合并多张色谱指纹图谱, 解决中药多元指纹图谱相似性评价难题, 提出一种多元色谱指纹图谱相似度计算方法. 用该方法先对各单元指纹图谱进行串行或并行象素级信息融合, 再对融合了各指纹峰信息的多元色谱指纹图谱进行相似度评价. 计算机仿真和复方丹参滴丸多元HPLC指纹图谱应用结果表明, 该法能够评价中药多元色谱指纹图谱相似度, 定量表征中成药产品批次间质量波动情况.     

HPLC指纹图谱相似度研究

将HPLC指纹图谱看作多维空间内的向量,利用向量夹角余弦的基本公式计算两个指纹图谱间的相似度。用7个不同品种的金银花提取物的HPLC指纹图谱对计算方法进行了检验,结果表明相似度能较好地定量评价指纹图谱间的相似性,可应用于中药质量控制。     

系统指纹定量法鉴别龙胆泻肝丸质量

建立龙胆泻肝丸(Longdanxiegan pill,LDXGP)的高效液相色谱(HPLC)指纹图谱,以系统指纹定量法评价其质量.采用RP-HPLC法,以宏定性相似度为参量对12批LDXGP进行聚类分析,确定用其中10批生成对照指纹图谱(RFP),以此RFP为标准用系统指纹定量法评价12批LDXGP质量.以龙胆苦苷为参照物峰,确定60个共有指纹峰,建立了LDXGP的HPLC指纹图谱.用系统指纹定量法鉴别出8批质量合格,1批含量明显偏高,1批含量明显偏低,2批化学成分数量和分布比例不合格.系统指纹定量法将整体定性分析和整体定量分析密切结合,是评价中药质量的有效方法.     

相似系统理论定量评价中药材色谱指纹图谱的相似度

研究相似系统理论定量评价中药材提取物液相色谱指纹图谱的相似度。以相似系统理论对栀子药材提取物高效液相色谱指纹图谱的相似度评价,发现相似系统理论对数据的差异比较敏感,而且相似度的计算结果能够反映样品的相对差异。相似系统理论可以作为一种评价中药色谱指纹图谱相似度的新方法。     

非线性化学指纹图谱技术用于菖蒲的鉴别及二元混合物中不同菖蒲含量的测定

采用非线性化学指纹图谱技术获得了石菖蒲、水菖蒲和九节菖蒲的特征指纹图谱,直观差异明显;将石菖蒲分别与水菖蒲和九节菖蒲进行二元掺杂,利用指纹图谱定量信息和各菖蒲含量拟合得到线性回归方程,相关系数0.9990.用该方程预测了二元混合物中不同菖蒲的含量,相对误差≤4.17%,RSD≤2.73%.非线性化学指纹图谱技术具有良好的重现性和特征性,同种菖蒲指纹图谱间系统相似度≥0.9989,不同种菖蒲指纹图谱间相似度≤0.9123.此外,该技术摒弃传统方法复杂的前处理过程,可高效、快速进行鉴别,从整体性角度对样本进行定性鉴别与定量分析.     

金银花的毛细管电泳指纹图谱研究

采用毛细管区带电泳法,以50 mmol/L硼砂(含20 mmol/L β-环糊精(CD),用磷酸调pH 8.0)为背景电解质,运行电压12 kV,紫外检测波长254 nm,重力进样15 s（高度8.5 cm）,建立了金银花药材水提取液的毛细管电泳指纹图谱(CEFP)。将13个不同产地的金银花药材供试液的CEFP进行比较,以电泳峰出现率100%计,确定金银花的共有指纹峰为18个。该CEFP具有较好的精密度和重现性,分离效能高且成本低廉。提出了指纹图谱宏观含量相似度R、投影含量相似度C和定量相似度P的概念,可从总体上评价药材化学组分的整体含量情况。从两个方面评价各产地药材与对照CEFP间的总体相似性,合格药材应具备以下两个条件：(1)代表化学成分分布相似性的定性相似度(S)≥0.90;(2)描述药材整体化学成分含量的定量相似度(R,C,P,Q)应在80%～120%。以此二类相似度指标控制金银花的质量,建立了指纹图谱评价的又一新方法。     

基于三波长融合谱的系统指纹定量法鉴定龙胆泻肝丸的真实质量

建立了龙胆泻肝丸(Longdanxiegan pill,LDXGP)三波长融合高效液相色谱(HPLC)指纹图谱,以系统指纹定量法全面鉴定LDXGP的质量。采用反相高效液相色谱法(RP-HPLC),运用多波长融合指纹图谱技术对色谱图进行处理,以黄芩苷为参照物峰,确立了63个共有指纹峰,以宏定性相似度为参量对12个厂家的12批LDXGP进行聚类分析,确定用其中10批生成对照指纹图谱(RFP),以此RFP为标准用系统指纹定量法评价12批LDXGP的质量。结果鉴别出9批质量完全合格,1批含量明显偏高,2批化学成分数量和分布比例不合格。基于多波长融合技术的系统指纹定量法是评价中药真实质量的可靠方法。     

Assessment of quality consistency in traditional Chinese medicine using multi‐wavelength fusion profiling by integrated quantitative fingerprint method: Niuhuang Jiedu pill as an example

Under the wave of the revival of traditional Chinese medicine, there is a quite imperative duty to study an integrated and comprehensive method of fingerprint data processing and analysis on the quality consistency of traditional Chinese medicine. So, we proposed six parameters from two aspects (qualitative and quantitative), three levels (biased to strong peaks, biased to weak peaks, no obvious bias), to comprehensively evaluate the similarity of the two fingerprints. On this basis, another five parameters were proposed to evaluate the integrated effects (consistency, volatility, and similarity). This method was applied to 22 batches of Niuhuang Jiedu pill samples. Next, a practical and convenient multi‐wavelength fusion method was designed to provide more information, and the generated fusion profilings were used for subsequent evaluation. The characteristics of the parameters were confirmed by correlation analysis. The results of both hierarchical clustering analysis and principal component analysis for raw data and standardized data were consistent with integrated quantitative fingerprint method results. At the same time, this method gave a reasonable explanation for abnormal and dissimilar samples. This work illustrated that the proposed method was particularly suitable for similarity analysis of fingerprints and capable of ensuring the quality consistency in traditional Chinese medicine.     

Capillary electrophoresis fingerprinting coupled with chemometrics to evaluate the quality consistency and predict the antioxidant activity of Sanhuang tablet as part of its quality control

A capillary electrophoresis fingerprint was constructed for Sanhuang tablet, a Chinese traditional patent medicine, that was commonly used in clinical practice, where the isosceles trapezoid method was first applied for the optimization of background electrolyte solution, and the resolution index was performed to assess the experimental conditions; furthermore, a novel linear quantitative fingerprint method was established for accurate qualitative and quantitative discrimination of the test samples from diverse commercial brands. The fingerprint analysis coupled with quantitative determination of two components was employed to elucidate that the quality consistency of the products was relatively good within one manufactory, but poor among different companies for the 30 batches of samples. In addition, the fingerprint–efficacy relationship between chemical components and antioxidant activity in vitro was investigated using partial least squares analysis, and the calibration and prediction of the antioxidant activity of the selected samples via fingerprint data were presented with the desired results. This work illustrates that the proposed fingerprint analysis based on linear quantitative fingerprint method can be applied for the quality evaluation of traditional Chinese medicine and herbal preparations as part of their quality control, and the constructed mathematical model is particularly suitable for depicting the fingerprint–efficacy relationship.     

指纹图谱法在参麦注射液质控中的应用

 中医药理论和实践要求综合评价中药的质量 ,指纹图谱法是对中药制剂进行综合宏观分析的可行手段之一 ,因此采用反相高效液相建立了参麦注射液的特征指纹图谱。条件 :Hypersil C18(4 6mmi d × 2 5 0mm ,5 μm)反相柱 ,流动相由水 (A)和乙腈 (B)组成 ,B的体积分数在 5 0min内由 5 %线性增长到 95 % ,流速为1 0mL/min ;紫外检测 ,波长为 2 0 2nm。 2 3个特征指纹峰与内标 (联苯 )的峰面积比作为指标 ,结合主成分分析 投影判别法比较了同一厂家不同批次产品和不同厂家同类产品的化学指纹差异。     

Monitoring quality consistency of Liuwei Dihuang Pill by integrating the ultraviolet spectroscopic fingerprint,a multi‐wavelength fusion fingerprint method,and antioxidant activities

Liuwei Dihuang Pill, a classical traditional Chinese medicine, has been widely used to treat kidney yin deficiency in China for hundreds of years. Little attention, however, has been paid to quality control methods for this formulation. In the present study, we aimed to establish a comprehensive and practical quality evaluation system for Liuwei Dihuang Pill. Thus, ultraviolet spectroscopic fingerprints and multi‐wavelength fusion fingerprints have been proposed for quality consistency evaluation of the popular patent. The similarity analysis of Liuwei Dihuang Pill samples was evaluated by systematic quantitative fingerprint method from qualitative and quantitative perspectives. The results showed that 26 batches of samples were classified into five grades for fusion fingerprints and three grades for ultraviolet spectroscopic fingerprints. Moreover, the fingerprint–efficacy correlation was also established and confirmed through the partial least squares model to visualize the antioxidant activity of Liuwei Dihuang Pill in vitro. In conclusion, these results indicated that integrating ultraviolet spectroscopic fingerprints and multi‐wavelength fusion fingerprints coupled with antioxidant activities provide a rapid and effective approach to monitor the quality consistency of Liuwei Dihuang Pill.     

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.     

基于水溶性浸出物及HPLC指纹图谱评价不同贮存年限半夏的品质

研究不同贮存年限半夏药材的浸出物,建立浸出物的HPLC特征指纹图谱,为半夏药材品质评控提供参考。浸出物测定方法采用药典法;HPLC指纹图谱的色谱条件:采用C_(18)色谱柱(150 mm×4.6 mm,5μm),以水–甲醇为流动相,梯度洗脱,流量为0.8 m L/min,检测波长为260 nm,柱温为25℃,进样体积为50μL。采用相似度评价及聚类分析技术揭示14批样品的相似性及差异性。14批半夏浸出物有12批合格,2批不合格。建立14批半夏浸出物样品的高效液相指纹图谱,确定了3个共有峰,共有峰保留时间的相对标准偏差小于2%,峰面积的相对标准偏差差异较大。1~#~7~#半夏样品有12个共有峰,共有峰保留时间的相对标准偏差小于1.5%,峰面积的相对标准偏差差异较大。各批次药材化学成分组成及含量均存在一定差异。以半夏浸出物数据与其高效液相色谱指纹图谱数据为基础,将指纹图谱相似度评价与聚类分析结合起来,用浸出物含量及评价软件测评结果对半夏品质进行综合评估,可以更精确地对半夏药材进行质量控制。     

Quality Assessment of Artemisia rupestris L. Using Quantitative Analysis of Multi-Components by Single Marker and Fingerprint Analysis

The chromatographic fingerprint of 14 batches of Artemisia rupestris L. samples were established in this study. The constituents of ten components in Artemisia rupestris L. were determined using quantitative analysis of multi-components by single marker (QAMS) and the external standard method (ESM). Due to their stability and accessibility, chlorogenic acid and linarin were used as references to calculate the relative correction factors (RCFs) of apigenin-C-6,8-pentoside-hexoside, apigenin-C-6,8-di-pentoside, luteolin, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, chrysosplenetin B, and sbsinthin, based on high-performance liquid chromatography (HPLC). The value calculated by QAMS was consistent with that of the ESM, and the reproducibility of RCFs was found to be reliable. In conclusion, simultaneous determination of the ten components by the QAMS method and chromatographic fingerprint analysis were feasible and accurate in evaluating the quality of Artemisia rupestris L. and can be used as reference in traditional Chinese medicine quality control.