高效液相法测定益母草中益母草碱含量研究

建立一种高效液相色谱(HPLC)检测鲜益母草及其制剂中益母草碱含量的方法。色谱柱为依利特Hypersil柱,流动相为0.00125 mol/L KH2PO4-CH3CN(87∶13,V/V),流速1 mL/min,柱温35℃,检测波长为276 nm。益母草碱的标准曲线回归方程是y=227 394x 1 292 605,r=0.9999(n=5)。该方法测定益母草及其制剂中益母草碱的含量,具有简便、准确及可靠的优点。     

运用RBF径向基神经网络识别基于小波变换的淫羊藿苷红外光谱指纹特征

运用RBF径向基神经网络对基于傅里叶变换红外光谱（FTIR）法、相关系数比对法及多层次小波变换所提取的淫羊藿药材样本有效成分指纹特征进行识别。样品包括药典选用的５个物种（淫羊藿Epimedium brevicornu Maxim.、箭叶淫羊藿E.sagittatum（Sieb. et Zucc.） Maxim.、柔毛淫羊藿E.pubescens Maxim.、朝鲜淫羊藿E.koreanum Nakai和巫山淫羊藿E.wushanense T.S.Ying）和其他物种共计250个样本。淫羊藿指标成分淫羊藿苷在原药材和甲醇提取物的红外光谱图上,具有较明显的1 259 cm-1特征峰,经典的HPLC法的定量分析结果也佐证了淫羊藿苷的含量与1 259 cm-1峰的位置具有很好的一致性。为此该峰可以用来作为判别各物种淫羊藿药材是否含有淫羊藿苷的重要依据。在此基础上,采用相关系数比对法和多层次的小波变换法消除了因淫羊藿苷含量低、吸收峰弱,信号和噪声大的问题,增强了RBF径向基神经网络的识别效果。初步建立了基于小波变换和RBF径向基神经网络淫羊藿原药材红外光谱快速识别淫羊藿苷指标成分的一种新方法。     

A comparative study of Mentha arvensis L. and Mentha haplocalyx Briq. by HPLC

We have developed a new method to simultaneously determine five marker compounds in Menthae Herba via HPLC/PDA – including hesperidin (1), rosmarinic acid (2), diosmin (3), didymin (4) and buddleoside (5). The newly developed method was successfully used to analyse for two species (Mentha arvensis L. and Mentha haplocalyx Briq.) of Menthae Herba, and the satisfactory results were obtained from the validation of developed method. The pattern analysis could greatly discriminate between M. arvensis L. and M. haplocalyx Briq. In conclusion, the proposed HPLC/PDA method is suitable for quality evaluation of Menthae Herba.     

Integrated Network Pharmacology Analysis and In Vitro Validation Revealed the Potential Active Components and Underlying Mechanistic Pathways of Herba Patriniae in Colorectal Cancer

Herba Patriniae (HP) are medicinal plants commonly used in colorectal cancer (CRC) patients. In this study, network pharmacology was used to predict the active components and key signaling pathways of HP in CRC. Patrinia heterophylla, one type of HP, was chosen for validation of the network pharmacology analysis. The phytochemical profile of Patrinia heterophylla water extract (PHW) was determined by UHPLC-MS. MTT, RT-PCR, and Western blot assays were performed to evaluate the bioactivities of PHW in colon cancer cells. Results showed that 15 potentially active components of HP interacted with 28 putative targets of CRC in the compound–target network, of which asperglaucide had the highest degree. Furthermore, the ErbB signaling pathway was identified as the pathway mediated by HP with the most potential against CRC. Both RT-PCR and Western blot results showed that PHW significantly downregulated the mRNA and protein levels of EGFR, PI3K, and AKT in HCT116 cells. Asperglaucide, present in PHW, exhibited an anti-migratory effect in HCT116 cells, suggesting that it could be an active component of PHW in CRC treatment. In conclusion, this study has provided the first scientific evidence to support the use of PHW in CRC and paved the way for further research into the underlying mechanisms of PHW against CRC.     

Differentiation of Herba Cistanches by fingerprint with high-performance liquid chromatography-diode array detection-mass spectrometry

Herba Cistanche (Rou Cong Rong in Chinese), dried succulent stems of Cistanche deserticola or C. tubulosa, is a famous Chinese herbal medicine and has been recorded in the Chinese Pharmacopoeia. In recent years, another two non-official species, C. salsa and C. sinensis have also been used as Herba Cistanche in some regions of China. To investigate the possibility of using these two non-official species as alternatives to the official species, a high-performance liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS) fingerprint method was developed to comparatively analyze the crude herbs of these four species. The fingerprint of C. deserticola, a historically certified species of Herba Cistanche, serves as 'standard pattern' for comparing the similarities with the other species by means of similarity and Principle Component Analysis. Additionally, 18 characteristic peaks in the fingerprints were identified by comparing their retention times, UV spectra and ESI-MS data with those of the reference substances and/or the data in the literatures. The comparative results demonstrate that the fingerprints of C. tubulosa and C. salsa possess high similarity to the standard pattern, suggesting that these two species may be used as alternative species; while that of C. sinensis has low similarity (0.053 correlation coefficient) to the standard pattern, indicating that it cannot be used as the substitute of the official herb. However, the varying fingerprint patterns among the samples of C. deserticola collected from various habitats illustrate that the quality consistency of crude herbs is still a problem worthy of serious concern.     

High-Performance Liquid Chromatographic Method for Determination and Pharmacokinetic Study of Chlorogenic Acid in the Plasma of Rats After Administration of the Chinese Medicinal Preparation Luying Decoction

A simple and sensitive high-performance liquid chromatographic method has been developed for determination of chlorogenic acid in rat plasma. Chlorogenic acid was extracted from plasma samples with methanol. HPLC analysis of the extracts was performed on a C₁₈ column (250 mm × 4.6 mm i.d., 5 µm particles). The mobile phase was acetonitrile −1% formic acid (9:91, v/v). The calibration plot was linear over the range 0.0420–2.10 µg mL⁻¹ and the lower limit of quantification was 0.0420 µg mL⁻¹. The method was reproducible and reliable with intra-day precision better than 8.2%, inter-day precision better than 9.1%, accuracy within ±8.3%, and mean extraction recovery above 84.4%. The validated method was successfully applied to pharmacokinetic studies of chlorogenic acid in rat plasma after administration of Luying decoction.     

Protective effects on ovalbumin-induced mouse asthma models and qualitative and quantitative analysis of multiple compounds in Gerberae Piloselloidis Herba

Gerberae Piloselloidis Herba is widely used to treat cough and asthma in China. However, its effects on allergic asthma as related to its chemical compositions have not been fully elucidated, and there is a scarcity of methods to determine multi-component contents for quality control. In this study, protective effects of Gerberae Piloselloidis Herba on ovalbumin-induced asthma models were investigated, while qualitative and quantitative analyses of multiple constituents in Gerberae Piloselloidis Herba were conducted by using an ultrahigh-performance liquid chromatography–Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry and an ultrahigh-performance liquid chromatography–photodiode array detection. The results showed that Gerberae Piloselloidis Herba could significantly mitigate asthma symptoms, reduce eosinophils counts in the bronchoalveolar lavage fluid, as well as decrease IgE, IL-5, and IL-13 concentration, and inflammatory cellular infiltration in lung tissues. A total of 51 compounds were tentatively identified, in which the content of 10 representative compounds was determined in 24 batches of Gerberae Piloselloidis Herba by using an ultrahigh-performance liquid chromatography method with good linearity, precision, repeatability, accuracy, and stability. This research presents a comprehensive strategy combining biological activity evaluation with chemical profiling, providing a useful and comprehensive reference for further application and quality control of Gerberae Piloselloidis Herba.     

Quantitative analysis of Cistanches Herba using high‐performance liquid chromatography coupled with diode array detection and high‐resolution mass spectrometry combined with chemometric methods

We aim to determine the chemical constituents of three species of Cistanches Herba using HPLC coupled with diode array detection and high‐resolution MS. Ten phenylethanoid glycosides were identified and further quantified as marker substances by HPLC coupled with diode array detection method. The separation was conducted using an Agilent TC‐C18 column with 0.1% formic acid and methanol as the mobile phases under gradient elution. The analytical method was fully validated in terms of linearity, sensitivity, precision, repeatability as well as recovery, and subsequently applied to evaluate the quality of 36 batches of Cistanche plants. The chemometric procedures (i.e., hierarchical clustering analysis and principal component analysis) were used to compare different species of Cistanches Herba, leading to successful classification of the Cistanche samples in accordance with their origins. In conclusion, this study provides a chemical basis for quality control of Cistanches Herba.     

Quality Evaluation of Taxilli Herba from Different Hosts Based on Simultaneous Determination of Multiple Bioactive Constituents Combined with Multivariate Statistical Analysis

Taxilli Herba (TAXH) is an important traditional Chinese medicine with a long history, dating from the Eastern Han Dynasty to the present times. However, the active constituents in it that parasitize different hosts vary, affecting its clinical efficacy. Given the complexity of the host origins, evaluating the quality of TAXH is critical to ensure the safety and effectiveness of clinical medication. In the present study, a quantitative method based on ultra-fast liquid chromatography tandem triple quadrupole mass spectrometry (UFLC-QTRAP-MS/MS) was established, which simultaneously determined the content of 33 active constituents, including 12 flavonoids, 4 organic acids, 12 amino acids, and 5 nucleosides in 45 samples. Orthogonal partial least squares discriminant analysis (OPLS-DA) was employed to classify and distinguish between TAXH and its adulterants, Tolypanthi Herba (TOLH). A hierarchical clustering analysis (HCA) was conducted combined with a heatmap to visually observe the distribution regularity of 33 constituents in each sample. Furthermore, gray relational analysis (GRA) was applied to evaluate the quality of samples to get the optimal host. The results demonstrated that TAXH excelled TOLH in quality as a whole. The quality of TAXH parasitizing Morus alba was also better, while those that were parasitic on Cinnamomum camphora and Glyptostrobus pensilis had relatively poor quality. This study may provide comprehensive information that is necessary for quality control and supply a scientific basis for further exploring the quality formation mechanism of TAXH.     

淫羊藿药材一些混淆物种的FTIR鉴别研究

淫羊藿是一味药用历史悠久的传统中药, 具有补肾壮阳之功效。《中国药典》规定使用五种来源的淫羊藿——淫羊藿Epimedium brevicornu Maxim.、箭叶淫羊藿E.sagittatum(Sieb. et Zucc.) Maxim.、柔毛淫羊藿E.pubescens Maxim.、朝鲜淫羊藿E.koreanum Nakai和巫山淫羊藿E.wushanense T.S.Ying,但这几种药材的鉴别特征描述比较简单,传统鉴定技术很难实现这些种类和常见混淆物种的鉴别。文章采用傅里叶变换红外光谱(FTIR)及其二阶导数谱对常见淫羊藿易混淆药材——淫羊藿与朝鲜淫羊藿,柔毛淫羊藿与保靖淫羊藿E.baojingense Q.L.Chen & B.M.Yang、星花淫羊藿E.stellulatum Stearn、竹山淫羊藿E.zhushanense K.F.Wu&S.X.Qian,柔毛淫羊藿与部分天平山淫羊藿E.myrianthum Stearn,箭叶淫羊藿与粗毛淫羊藿E.acuminutum Franch.共4对易混淆药材进行鉴别研究。根据不同物种淫羊藿药材红外光谱特征的不同,基本上实现常见易混淆物种的快速鉴别。该方法有助于解决淫羊藿药材物种混乱问题。