基于牛舌草指纹图谱的多指标成分相对定量与鉴别方法

色谱指纹图谱法在中草药产地鉴别或中成药鉴别中发挥着重要作用,然而色谱峰强度和位置易受目标物提取和分离条件的影响,发展更加准确的指纹图谱具有重要意义。该文以色谱峰中某一个常见组分（芦丁）作为内标,分析其他组分的相对含量,以相对含量作为指纹图谱信息,结合化学模式识别可以精确给出产地的相似度。以牛舌草的液相色谱指纹图谱指纹峰为例,以其中芦丁的组分峰作为基准峰,其他指纹峰以芦丁为参照,计算每个指纹峰以芦丁计的含量,建立了牛舌草的指纹图谱。将获得的相对含量指纹图谱采用系统聚类分析和主成分分析进行化学模式识别研究,实现了不同产地牛舌草的区分。该方法建立了牛舌草多指标成分的相对定量与鉴别方法,既节省资源,又具有可操作性,对于其他中药或中成药的质量控制具有一定的借鉴意义。     

三波长融合指纹图谱结合6组分定量和主成分分析评价银翘解毒片的质量

建立了三波长融合高效液相色谱指纹图谱,并结合6组分定量和主成分分析(PCA)评价25批银翘解毒片的质量一致性。采用反相高效液相色谱法(RP-HPLC)分别于230、279、327 nm下检测。运用多波长融合指纹图谱技术建立银翘解毒片三波长融合指纹图谱,采用系统指纹定量法对其进行定性和整体定量评价。结果鉴别出25批银翘解毒片样品完全合格且区分良好。同时测定6组分含量,与指纹图谱结合,从整体和局部角度评价银翘解毒片质量。此外,运用PCA法对融合指纹图谱进行分析,通过主成分得分图可以明显区分来自两个厂家的25批银翘解毒片样品。方法综合性较强且有效,为科学评价与有效控制银翘解毒片的质量提供了可靠的参考。     

应用电化学指纹图谱优化黄连水提取工艺的研究

以黄连提取液中的总成分为定量指标,应用电化学振荡技术获取不同提取工艺的电化学指纹图谱.根据电化学指纹图谱特征参数诱导时间来考察其水溶性有效成分含量的高低,与高效液相色谱含量测定方法对照,验证了该方法的可靠性,确定黄连提取的最佳工艺为:采用12倍量的水,提取1.5 h,提取2次.结果表明利用电化学指纹图谱的诱导时间优化中药水溶性有效成分的提取工艺是可行的.     

葛根芩连配方颗粒标志性成分含量分析及指纹图谱研究

采用反相高效液相色谱法-二极管阵列检测器,以甲醇-水(内含1%三乙胺、1%乙酸,用磷酸调pH至3)为流动相,对中药复方制剂葛根芩连配方颗粒中3种主要标志性成分的含量同时分析测定,并采用相同的流动相对其指纹图谱进行研究。实验结果显示,3种主要标志性成分葛根素、黄芩苷和小檗碱在含量分析测定条件下,各组分分离良好,相关系数均为0.9999,线性范围分别为0.042~1.05、0.13~3.25和0.060~1.50μg,加样回收率在95%~106%之间;在指纹图谱研究中,以黄芩苷为参照物峰,标定了32个共有峰,分析的11批葛根芩连配方颗粒与共有模式之间具有良好的相似性,相似度均在0.9以上。建立的含量测定及指纹图谱分析方法灵敏度高、专属性强,可用于葛根芩连配方颗粒的质量控制。     

基于化学指纹图谱和抗血小板聚集效价的丹参质量评价

通过化学分析和生物活性评价考察丹参药材的品质差异,探讨丹参抗血小板聚集生物活性的主要贡献成分.采用高效液相色谱(HPLC)技术建立丹参药材HPLC指纹图谱,以抗血小板聚集相对效价作为指标,评价不同产地不同批次丹参药材的品质差异,构建基于化学表征及生物效价测定的评价模式.结果表明,不同批次丹参药材的HPLC指纹图谱相似度很高(相似度0.930~0.998),而其抗血小板聚集相对效价相差10倍,提示化学指纹图谱难以反映丹参的活性和质量差异.通过化学指纹图谱与抗血小板聚集生物效价进行谱效相关分析,筛选出与生物活性相关系数大于0.5的6个色谱峰:二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ、丹参酮ⅡA及2个未知化合物.对上述4种已知化合物单体进行活性验证发现,隐丹参酮的抗血小板聚集活性最强,而其它3种丹参酮类化合物几乎没有体外抗血小板聚集活性.进一步比较丹参中高含量成分丹酚酸B与低含量成分隐丹参酮的活性贡献,结果表明,两者的活性贡献基本相当,说明隐丹参酮是丹参中低含量高活性成分,对评价丹参质量具有重要贡献度.     

人工蚕蛹虫草和冬虫夏草水溶性成分的相对归属分析

为对相近中药的化学指纹图谱进行比较研究,充分利用联用色谱所得的光谱、色谱信息与光谱相关色谱方法,分析了蚕蛹虫草和冬虫夏草的水溶性成分的异同,实现了蚕蛹虫草和冬虫夏草水溶性成分化学指纹图谱中相应化学组分的归属分析。其中,蚕蛹虫草和冬虫夏草有5个共有组分,分别有4个和6个相异组分。实验表明：该方法可对复杂中药成分及其指纹图谱进行快速比较分析。     

白术组分拆分及组间差异度评价方法的建立

通过多种分离方法联合应用,将白术化学成分拆分成5个部分,各拆分组分组内指纹图谱相似度评价表明,拆分工艺稳定、重现性良好。采用差异度表征中药化学拆分组分间的不交叉程度。利用主成分分析、聚类分析及夹角余弦、平方欧式距离和生药峰面积交叉分析法对白术化学拆分组分进行不交叉的定性与定量分析,表明白术化学拆分组分组间差异度不低于85%。其中生药峰面积差异度分析法是在中药指纹图谱基础上建立的表征中药拆分组分差异度的新方法。通过与不同分析方法比较,发现其较适合中药药效或药性物质研究中的拆分组分间不交叉程度的评价。     

中药组方指纹图谱研究方法和思路

中药组方就是将单味中药按照一定的规律配伍成方,因剂量配伍差异会导致组方理化性质的改变,从而导致组方功效强弱、性质方面发生变化,甚至产生新的化学成分。中药组方鉴别与质量控制一直是困扰中药工作者的一个难题。指纹图谱技术具有宏观、准确、稳定等优点,已被广泛用于中药领域。中药组方指纹图谱是解决中药合理组方配伍的最有效手段,其对中药质量控制已经得到普遍关注和重视。由于指纹图谱具有整体定量功能和特征性鉴别作用,对于成分复杂的中药组方能够全面地定性与定量评价。本文就中药组方指纹图谱的研究意义与目的进行了综述,并详细阐述了组方指纹图谱的研究方法与思路,以此为中药及其组方指纹图谱研究提供参考。     

具有油水分配系数测算功能的中药指纹图谱

建立了一种能同时测算中药中主要成分的油水分配系数Pow的功能化中药指纹图谱。采用微乳液电动色谱,以中药女贞子为例,对运行缓冲液浓度、pH值、十二烷基硫酸钠浓度等参数进行了优化。利用SPSS程序对磺胺等从女贞子中提取的6种标准化合物组分的迁移时间tm与其log Pow文献值进行非线性拟合,得到女贞子提取组分的标准方程线性关系良好（r=0.9880）。在相同的实验条件下测得化合物的log Pow与文献报道值较好的符合,证明该方法可靠。获得了能代表组分疏水化学特征的中药指纹图谱,测得女贞子的主要有效成分齐墩果酸的log Pow为3.63。该指纹图谱较为可靠地提供了女贞子中各种成分的log Pow,直观地反映出各组分油水分配性质的分布,对中药有效成分提取溶剂的选择、剂型的设计及制剂工艺的改进具有指导意义。     

山东不同产地丹参的HPLC指纹图谱-化学模式识别研究

建立RPHPLC指纹图谱-化学模式识别评价丹参质量的方法.利用RPHPLC法测定不同产地丹参药材的指纹图谱及丹参酮ⅡAt和丹参酮的含量,采用主成分分析、系统聚类分析和逐步判别分析对指纹图谱信息进行化学模式识别研究.在主成分分析的基础上,以前4个主成分为聚类分析的指标进行系统聚类分析,取阈值为8时,所有样品可被分为4类;并建立了相应的判别函数,回判准确率100%;以此为依据,初步建立了丹参化学模式识别的评价方法.     

循环制备液相色谱分离芳香新塔花中的化学成分

建立了交替循环和直接循环液相色谱相结合的方法用于制备芳香新塔花中的化学成分.芳香新塔花样品经溶剂提取、柱色谱和中压制备色谱初步分离后得到芳香新塔花的不同馏分.以甲醇-水为流动相,利用双柱交替循环法对组分进行分离,同时,流动相经恒流泵循环输入色谱柱.以馏分Ⅰ和馏分Ⅱ为例,在混合循环模式下分离得到5个化合物.通过核磁共振对其进行鉴定,确定分别为乔松素-7-O-芸香糖苷、白杨素-7-O-芸香糖苷、金合欢素-7-O-芸香糖苷、云杉素和原儿茶酸.实验结果表明,该制备方法分离效率高,节省流动相,是分离天然产物的有效手段.     

制备型高效液相色谱法分离纯化川西獐牙菜提取物中的龙胆苦苷

龙胆苦苷(GPS)是龙胆类药材及其相关制品质量控制的指标成分。本研究利用制备型高效液相色谱从川西獐牙菜提取物中分离纯化龙胆苦苷对照品。对制备色谱的流动相组成、流速、进样量和检测波长等制备参数进行了优化。采用的色谱柱为C18柱(200 mm×50 mm, 5 μm),流动相为甲醇和0.1%乙酸水溶液(体积比为30:70),流速为75 mL/min,检测波长为254 nm,进样体积为500 μL。在30 min的运行时间内,龙胆苦苷与其他干扰成分得到了很好的分离,产品纯度达到了99%以上。此方法具有快速高效、产品纯度高的特点,可用于制备龙胆苦苷对照品和对龙胆苦苷制品的质量控制。     

Improved quality assessment of proprietary Chinese medicines based on multi‐chemical class fingerprinting

Our present study constitutes the first successful attempt to employ eight distinctive chemical groups of compounds for the quality evaluation of a complex traditional Chinese herbal medicine (TCHM) material: BaZhen YiMu (BZYM). Due to the complexity of its matrix, which is composed of nine different herbal ingredients, five representative chemical groups encompassing representative bioactive markers were initially chosen as targets for the quality assessment of this preparation. Furthermore, with the aid of LC‐ESI‐MS, three additional chemical groups were characterized. In summary, a total of nineteen markers belonging to eight different chemical groups were selectively displayed in the chromatographic fingerprint of BZYM preparation. With this fingerprint, the overall quality of any BZYM preparation can be comprehensively authenticated. The chromatographic separation was performed on an HP C₁₈ AQ column with a gradient elution of ACN and aqueous solution containing 0.1% phosphoric acid at the optimal detection wavelength of 230 nm. The established method was rigorously validated with respect to the ICH guidelines and represents the most extensive and facile HPLC quality control technique for this formulation. Compared with the conventional method of using a single or only a few markers of the same chemical group, this technique provides a new dimension for TCHM quality control.     

Solid-phase microextraction of hop volatiles. Potential use for determination and verification of hop varieties

The composition of hop essential oil is an important tool for evaluation of hop quality. As each hop variety has a typical essential oil pattern (fingerprint), hop oil analyses can be used to distinguish between hop varieties. The headspace solid-phase microextraction (SPME) method as described in this contribution is a simple sample preparation technique and represents an alternative procedure for essential oil fingerprint determination. Different SPME parameters (extraction temperature, extraction time and sample mass) were studied and the results were compared with those obtained by the routine distillation method. It is shown that SPME results can be used for determination and verification of varieties grown in Slovenia by means of principal components analysis.     

高效制备液相色谱法从荷叶中分离制备黄酮类化合物

采用高效制备液相色谱法从荷叶(Nelumbo nucifera Gaertn)中分离制备荷叶黄酮类化合物。用60%乙醇回流提取荷叶，粗提液浓缩后经D-101柱及聚酰胺柱色谱分离，再在Symmetry PrepTM C18柱上分离,以水-乙腈为流动相进行梯度洗脱（流速为5.0 mL/min）,得到了3种黄酮类化合物。经紫外光谱、红外光谱、核磁共振及质谱分析,确定该3种物质分别为金丝桃苷、异槲皮苷和紫云英苷。所制备的3种化合物的纯度都在97%以上,其中紫云英苷为首次从荷叶中分离得到。     

制备色谱法分离纯化高良姜黄酮中高良姜素和山柰素

建立了制备型高效液相色谱(Prep-HPLC)分离高良姜黄酮中高良姜素和山柰素的方法。高良姜黄酮经HPD-600树脂吸附洗脱纯化后,采用Prep-HPLC分离高良姜黄酮中高良姜素和山柰素。制备色谱条件:流动相为甲醇-0.6%(v/v)乙酸水溶液(58:42, v/v),柱温为常温,流速为7.0 mL/min,检测波长为360 nm,进样量为700 μ L,上样质量浓度为10.0 g/L。分离的单体由质谱和核磁共振氢谱、碳谱鉴定确证为高良姜素和山柰素,HPLC外标法定量,纯度分别为99.5%和99.7%。该方法分离效果好、高效、低毒,可用于高良姜中高良姜素和山柰素的分离制备。     

Slurry sampling electrothermal atomic absorption spectrometric determination of chromium after separation/enrichment by mercaptoundecanoic acid modified gold coated TiO2 nanoparticles

In this study, a novel preconcentration/separation technique based on the slurry analysis of chromium loaded on mercaptoundecanoic acid modified TiO₂ core-Au shell nanoparticles prior to its determination by electrothermal atomic absorption spectrometry was described. For this purpose, at first, TiO₂ nanoparticles were coated with gold shell and then modified with mercaptoundecanoic acid (MUA). Cr (III) was collected on the prepared sorbent by conventional batch technique. After separation of liquid phase, slurry of the sorbent was prepared and directly introduced into graphite furnace of atomic absorption spectrometry. By this way, all drawbacks due to elution procedure were eliminated. Optimum conditions for quantitative sorption and preparation of the slurry were investigated. The chromium in certificated sea-water and spiked drinking water was recovered in the range of 95% confidence level. The proposed technique was fast and simple as well as the risks of contamination and loss during elution were low. The limit of detection (3σ, N = 10) was 0.34 μg L^{− 1}.     

基于主成分分析的骨碎补药材乙醇和环己烷提取物的高效液相色谱指纹图谱及指标成分的定量分析

建立了骨碎补药材乙醇和环己烷提取物的高效液相色谱(HPLC)指纹图谱,并利用主成分分析法(PCA)对指纹图谱进行统计分析,以各主要色谱峰的保留时间和峰面积为变量得到score图和loading图。在score图和loading图中,骨碎补的正品和非正品可明显区分,且揭示出对此区分贡献最大的4个潜在指标成分,其中已知成分为柚皮苷、新北美圣草苷和E-4-O-β-D-葡萄糖酰咖啡酸。同时测定了这3种成分在19批正品和非正品骨碎补药材中的含量,其中10批骨碎补药材正品中3种成分的含量为: 柚皮苷6.36～10.1 mg/g,新北美圣草苷5.14～9.21 mg/g,E-4-O-β-D-葡萄糖酰咖啡酸1.87～3.19 mg/g。该方法更全面地反映了药材的化学成分信息,并能从定性和定量两方面控制骨碎补药材的内在质量。     

Development of a method using high‐performance liquid chromatographic fingerprint and multi‐ingredients quantitative analysis for the quality control of Yangxinshi Pian

A simple and reliable method of high‐performance liquid chromatography with diode array detection method was developed for fingerprint analysis and simultaneous determination of six compounds including puerarin, salvianolic acid B, berberine hydrochloride, palmatine chloride, dehydrocorydaline, and icariin in the Chinese medicine preparation Yangxinshi Pian. The separation was performed on an Agilent Eclipse XDB‐C₁₈ reserved‐phase column (250 mm × 4.6mm I.D., 5 μm) using gradient elution with 50 mmol/L monopotassium phosphate aqueous solution and methanol as mobile phase at a flow rate of 1.0 mL/min. The column operating temperature was set at 30°C, and the detection wavelength was 280 nm. The method was validated by linearity, precision, accuracy, stability, and recovery. For fingerprint analysis, 25 peaks were selected as the common peaks, and four kinds of similarities including cosine similarity (S), ratio of similarity (S′), projection content similarity (C), and content similarity (P) were applied to evaluate the quality consistency of different batches of Yangxinshi Pian. The results showed that the developed method was an efficient tool for quality evaluation of Yangxinshi Pian.     

Determination of phthalic,terephthalic, and isophthalic acids in the presence of each other by gas chromatography

A procedure was developed for the simultaneous determination of phthalic acid isomers; the procedure includes the preparation of diisoamyl esters by reaction with isoamyl alcohol in a benzene solution in the presence of sulfuric acid with the distillation of an azeotrope mixture of benzene with water, the neutralization of acid with triethylamine, and the separation of esters by gas-liquid chromatography on a steel column (1 m × 3 mm) filled with Chromaton N-AW-HMDS with 5% Apiezon L in the temperature programming mode with flame-ionization detection. The time of separation is 13 min. The time of the sample preparation step is 2–2.5 h. The lower determination limit is 20 ng. The procedure allows the determination of phthalic acids in the concentration range from 0.02 to 2 mg/L in liquid samples and from 0.03 to 3 mg/m³ in air.