Fingerprint Analysis of Codonopsis Radix by HPLC Coupled with Chemometrics Analysis

Objective To establish a validated high performance liquid chromatography(HPLC)for the fingerprint analysis of Codonopsis Radix and for the determination of lobetyolin.Methods HPLC coupled with diode array detection method was employed to establish the fingerprint profile and quantitative determination of lobetyolin in Codonopsis Radix.Principal component analysis method was employed to analyze the 52 Codonopsis Radix samples.Results The reference chromatogram was generated with 25 common peaks showing good separation from adjacent peaks.Conclusion Statistical analysis of the obtained data demonstrates that the developed HPLC fingerprint combined with chemometric is a reliable method for the similar evaluation and quality assessment of Codonopsis Radix,and other traditional Chinese herbs.     

Quality Evaluation of Astragali Radix Products by Quantitative Analysis of Multi-components by Single Marker

Objective To develop a quantitative analysis of multi-components by single-marker(QAMS)method for the simultaneous determination of eight components in Astragali Radix products,and to examine the feasibility of using the method among the different dosage forms and between two different types of compounds.Methods Eight main effective components,campanulin,genistin,ononin,calycosin,genistein,formononetin,methylnissolin,and astragaloside IV were selected as analytes for the quality control of Astragali Radix products.Calycosin was selected as the internal reference substance,the content of which was determined by external standard method;the relative correction factors(RCFs)of campanulin,genistin,ononin,genistein,formononetin,methylnissolin,and astragaloside IV were calculated.In total,twelve Astragali Radix specimen in decoction pieces,as well as in two different dosage forms,such as granule and oral liquid products,were used for the quality control by both methods of external standard and QAMS.The validity of the QAMS method was evaluated by comparison on the quantitative results of the two methods.Results These RCFs were obtained with good reproducibility(RSD<6.5%)by using ultra high performance liquid chromatography coupled with diode array detector under various chromatographic conditions.Meanwhile,no obvious differences(RSD<3.98%)were found in the quantitative results of the seven components in twelve samples of Astragali Radix products determined by the two methods.Conclusion QAMS is a reliable and feasible method in determining the components in products of Astragali Radix.     

Quality Evaluation of Astragali Radix based on DPPH Radical Scavenging Activity and Chemical Analysis

Objective To assess the quality of Astragali Radix from different areas based on the biological evaluation and chemical analysis. Methods The bioassay method of 1,1-diphenyl-2-picryl-hydrazyl(DPPH) radical scavenging activity for Astragali Radix was established. The parameters of DPPH assay including sample extraction time, reaction time, repeatability, and stability were detected. Furthermore, a method of HPLC-MS was developed to simultaneously determine calycosin-7-O-glucoside, ononin, formononetin, and astragaloside IV in Astragali Radix samples. And the total flavonoids and total saponins were detected by spectrophotometry. The relationship between DPPH evaluation and chemical analysis was studied by Pearson correlation analysis. Results Twelvebatches of Astragali Radix from different origins showed a wide range of DPPH radical scavenging activities(IC50 = 1.395-9.894 μg/mL). Based on DPPH assay, Sample 10 derived from Inner Mongolia Autonomous Region(IC50 = 1.395 μg/mL) showed the best quality of all samples. Chemical analysis showed that different compounds selected as indices would cause different results for quality evaluation. Pearson correlation analysis revealed that the contents of total flavonoids(P = 0.032), calycosin-7-glucoside(P =0.035), and astragaloside IV(P = 0.010) were positively correlated with DPPH radical scavenging activity. Conclusion Except for chemical analysis, DPPH radical scavenging activity can be used as a good alternative to assess and control the quality of Astragali Radix.     

Effects of processing technology on essential oil and pharmacological action of frankincense

Background:According to the traditional Chinese medicine theory,the common processing methods for Ruxiang(frankincense)include stir-frying processing and vinegar processing.Methods:With network pharmacology and thermal analysis methods,we selected and identified the main and irritating ingredients,established the pyrolysis characteristic parameters for identifying frankincense quality,quantified the starting and optimum temperature limits for processing frankincense,and analyzed the influences of processing adjuvants(vinegar)on combustion pyrolysis characteristics of frankincense.By applying Fourier transformation infrared spectrometry,high performance liquid chromatography,gas chromatography and UV-visible spectrometry,we evaluated the processing technology developed in our study of frankincense and its processed products.Results:Based on network pharmacology,we can find that the classical compounds of essential oil are the main pharmacodynamics components of frankincense possessing the anti-inflammatory,analgesia,anti-tumor,anti-ulcer and ant-inflammatory bowel disease effects.The pyrolysis combustion rate peaks of frankincense at 285.27±7.05℃and 476.99±13.46℃were the characteristic peaks used to identify frankincense quality.The specific temperature of slow-fire processing,including the vinegar processing and stir-frying processing,was 148.72±5.09℃.Heating for 5.67±0.98 min after reaching processing temperature was needed for processing frankincense with vinegar.The kinetics analysis showed that both frankincense and its vinegar mixture exhibited good linear relationships.The feasibility and practicability for our processing technology were verified through the determination of medicinal compounds by Fourier transformation infrared spectrometry,high performance liquid chromatography,gas chromatography and UV.Conclusion:Frankincense essential oils were the main active components of frankincense,which also can be taken as the internal chemical control indexes to judge the quality control and to optimize the processing technology of frankincense.The pyrolysis combustion rate peaks of frankincense at 285.27±7.05℃and 476.99±13.46℃were the characteristic peaks that can be used to identify frankincense quality.The optimum processing conditions of vinegar-processed frankincense with were as follows:with vinegar,20%;heating temperature,148.72±5.09℃;and heating time,5.67±0.98 min.Under these conditions,frankincense essential oil could cause a moderate effect on combustion pyrolysis characteristics to exert its corresponding pharmacological effects.     

Comparative chemical characters of Pseudostellaria heterophylla from geographical origins of China

Objective: Pseudostellaria heterophylla has been paid more attention in recent years, mainly as a medicine food homology plant. The content determination of P. heterophylla is not specified in the Chinese Pharmacopoeia (version 2020). The environmental conditions in different production areas could exert an influence on the quality of P. heterophylla. The purpose of this study is to discriminate P. heterophylla collected from different geographical origins of China. Methods: In this study, the content of polysaccharide in 28 batches of P. heterophylla was determined using phenol-sulfuric acid. HPLC fingerprints were established under optimised HPLC-PDA methods. Subsequently, the similarity analysis (SA) and the quantification of heterophyllin B were analyzed. The metabolites of P. heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and orthogonal PLS-DA (OPLS-DA) were performed based on all peak areas. Results: The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas, which varied significant from different origins. While the content of heterophyllin B in Anhui and Jiangsu was high. The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990, and the characteristic map can be used to identify and evaluate the quality of P. heterophylla. The samples from Fujian, Guizhou, Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA, PLS-DA, HCA, OPLS-DA, indicating that their metabolic compositions were significantly different. Ultimately, a total of 15 metabolites which were filtrated by a VIP-value > 1 and a P-value ＜ 0.05 associated with the separation of different origins were identified. Conclusion: HPLC fingerprint was established to evaluate the quality and authenticity of P. heterophylla. The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions. A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P. heterophylla from different origins in China for the first time. Overall, this study provides insights to metabolomics of P. heterophylla and supplies important reference values for the development of functional foods.     

Simultaneous Determination of Saponins in Dripping Pills Made from Astragali Radix and Panax notoginseng by UPLC-ELSD

Objective To develop a simple and fast method for removing polyethylene glycol(PEG) and simultaneous determination of fives saponins, i.e. astragaloside IV, notoginsenoside R_1, ginsenoside Rg_1, ginsenoside Rb_1, and ginsenoside Rd in dripping pills made from Astragali Radix and Panax notoginseng. Methods The extraction method was based on a liquid-liquid extraction using water-saturated n-butanol and the quantitative determination was based on ultra-performance liquid chromatography coupled with evaporative light scattering detection(UPLC-ELSD). The chromatographic analysis was performed on an Acquity UPLC HSS T3 column(100 mm × 2.1 mm, 1.8 μm) with a gradient elution of acetonitrile-0.1% formic acid aqueous solution within a runtime of 15 min. Results Compared to different methods, the proposed method could remove the interference of PEG in formulation. And the calibration curves showed good linearity during the test ranges. The method was validated for limits of detection and quantification, precision, and reproducibility. The recoveries were within the range of 96.87%-99.97%. In addition, the verified method was firstly applied to determination of the five active ingredients in Qishen Yiqi Dripping Pills(QYDP) simultaneously.Conclusion The contents of five active ingredients are stable and homogeneous in QYDP, which indicates that the method could be readily utilized as a quality evaluation method for this traditional Chinese medicine dripping pills made from Astragali Radix and Panax notoginseng.     

Identification and activity evaluation of Astragalus Radix from different germplasm resources based on specific oligosaccharide fragments

Objective: Based on trifluoroacetic acid(TFA) hydrolysis, polyacrylamide gel electrophoresis(PAGE) and high performance thin layer chromatography(HPTLC) analysis, the carbohydrate responsible for immunomodulatory activity are used as quality indicators for Astragalus Radix(AR).Methods: In this study, 24 batches of AR from different germplasm resources were selected as the research object, and AR polysaccharides were extracted. PAGE and HPTLC methods were used to analyze the partial acid hydrolyzate of AR polysaccharides and obtain a series of saccharide fingerprints. The data were analyzed by principal component analysis to obtain the difference between AR from different germplasm resources.Results: The results showed that trisaccharide and tetrasaccharide could be used as differential fragments to distinguish AR of different cultivation methods;Disaccharides and trisaccharides can be used as differential fragments to distinguish different species of AR. The immunological activity analysis of the specific oligosaccharide fragment of AR showed that the specific oligosaccharide fragment of AR could promote the secretion of TNF-α, IL-1β, IL-6, and NO in THP-1 cells in a concentration-dependent manner.Conclusion: Both PAGE and HPTLC methods can be used to evaluate AR from different germplasm resources. This study laid the foundation for the quality evaluation of AR medicinal herbs.     

Toxicity Classification and Detoxification Strategies of Chinese Materia Medica

In recent years, more and more poison incidents of Chinese Materia Medica (CMM) were reported by China and other countries, which made people to doubt about the safety of CMM, especially for the toxic CMM. In this review, the toxicity of CMM and the toxicity classification of CMM were introduced. And traditional TCM theory and methods particularly used for control and attenuation of CMM’s toxicity were described in detail. The traditional processing technology and CMM drug combination were most important detoxifying measures used in CMM formulation and clinical practice. Finally, some factors that have influence on occurrence of CMM’s toxicity were discussed such as different species, producing technology and so on.     

The riddles of number nine in Chinese medicine processing method

The“nine cycles of steaming and shining”,“nine making”,“nine turns”and“nine cycles of calcining and quenching”methods that are recorded and used since ancient times are merely one aspect of the unique processing methods of traditional Chinese medicine.Inducing the Chinese medicine processing method“nine cycles of steaming and shining”from historical review and summarizing the practical wisdom based on the canonical aspects of traditional Chinese medicine and the experiences of ancient Chinese medicine sages to promote the new development of traditional Chinese medicine.After the long-term and multiple“nine”processing,the materials of traditional Chinese medicine exhibit significant beneficial changes in terms of taste,efficacy,and chemical composition contents,thus emphasizing that Chinese medicine processing plays a significant role in their efficacy enhancement and toxicity reduction.Heshouwu(Polygoni Multiflori Radix),Dihuang(Rehmanniae Radix),Huangjing(Rhizoma Polygonati),Dahuang(Radix et Rhizoma Rhei),and Xixiancao(Herba Siegesbeckiae)are representatives of Chinese medicinal materials prepared using the“nine-system”processing method.This review discovers the aim and the molecular mechanism of“nine”processing of the abovementioned herbs from the viewpoints of modern pharmacochemistry and pharmacology to provide a theoretical support for the“nine”processing method of traditional Chinese medicine and to promote the international market of traditional Chinese medicine.     

Spectrum-Toxicity Correlation Study Revealed the Influence of the Nine-Time Steaming and Sun Drying Method on Hepatotoxic Components of Polygoni Multiflori Radix

Objective: Polygoni Multiflori Radix (PM) is a traditional herbal medicine with repeated reports of liver injury events in recent years. We wondered whether the classical processing method, namely, nine-time steaming and sun drying (NSSD), had toxicity-attenuating effects on PM and the relationships between toxicity and times of processing, as well as with the alteration trends of its compounds. Materials and Methods: The chemical fingerprints of different PM extracts were developed using ultra-high-performance liquid chromatography. The spectrum-toxicity correlation between the chemical fingerprints and hepatocellular toxicity was analyzed with multiple correlation analysis. Results: The results suggested that the hepatotoxicity of NSSD processing products markedly decreased with the repeated steaming and sun drying, which was obviously superior to the product processed by the modern method. Comprehensive analysis revealed that the contents of cis-stilbene glycoside and emodin-8-O-β-D-glucoside related to liver injury susceptibility were reduced with the times of NSSD processing, which was consistent with the decreased trend of hepatocellular toxicity. After the five times of NSSD, the contents of them as well as the hepatotoxicity of PM were steady. Moreover, we found that the contents of catechin and physcion declined rapidly after the one time of NSSD and then remained stable until the nine times of NSSD. Based on the fact, they could be utilized to indicate whether PM products were processed by steaming and sun drying. Conclusions: This paper confirmed that the NSSD had a good influence on the toxicity attenuating to PM and found four compounds which could apply for the quality control of PM.     

基于AHP-熵权法结合色差原理评价不同辅料大米对米炒党参饮片的影响

目的 研究不同地区、不同种类的15批大米理化性质、米炒党参粉末颜色值与米炒党参主要有效成分含量的相关性,探究不同大米对米炒党参饮片质量的影响。方法 测定大米理化性质,利用色度仪对不同米炒制的党参粉末颜色值进行测量,采用紫外可见分光光度计对党参多糖进行定量,HPLC法测定党参炔苷、5-羟甲基糠醛含量,并测定醇浸出物含量,采用AHP-熵权法结合多元相关分析评价不同大米炒制的党参饮片质量。结果 在相同的炮制条件下,不同大米炒制的党参饮片质量均符合《中国药典》2020年版要求,但饮片内在质量存在一定差异。相关性分析结果显示米炒党参样品中党参多糖含量与颜色值L^*、a^*、E_ab^*及大米的过氧化氢酶活动度、直链淀粉含量呈正相关,党参炔苷含量与大米过氧化氢酶活动度成正相关,醇浸出物含量与颜色值L^*、a^*、b^*、E_ab^*及大米水分含量、直链淀粉含量均呈正相关。结论 不同地区、不同种类的大米对米炒党参饮片内在质量产生不同影响,米炒党参饮片主要有效成分含量与颜色值、大米理化性质有一定的相关性。     

干燥方法对何首乌块根中多元功效物质转化的影响

目的 研究不同干燥方法对何首乌药材中二苯乙烯苷类和蒽醌类活性成分转化的影响,为何首乌药材产地干燥加工适宜方法的建立提供科学依据。方法 以新鲜野生何首乌块根为材料,分别采用阴干、晒干、烘干、冷冻干燥、微波干燥、远红外干燥6种干燥加工方法处理,采用HPLC法同时测定经各种干燥方法制备的样品及不同烘干时间样品中二苯乙烯苷、游离型蒽醌类和结合型蒽醌类化学成分的量。结果 经6种不同干燥方法处理的样品中,二苯乙烯苷的量由高至低依次为晒干＞远红外干燥＞冷冻干燥＞阴干＞烘干＞微波干燥;阴干法和烘干法更适宜于结合型蒽醌类成分转化为游离型蒽醌类成分;采用远红外干燥的样品,其结合型蒽醌的量最高。主成分分析综合评价显示,经远红外干燥法制备的何首乌药材其综合评价指数明显高于其他干燥方式。结论 远红外干燥法为何首乌药材适宜的产地干燥加工方法。     

基于熵权法的灰色关联法-TOPSIS法对不同产地三七及其炮制品质量的评价研究

目的 利用熵权法结合灰色关联法-TOPSIS法建立三七Panax notoginseng质量评价模型,评价不同产地三七及其炮制品质量。方法 以水分、醇浸出物、皂苷含量、多糖得率、多糖总糖含量、糖蛋白含量及糖醛酸含量作为检测指标,以熵权法计算权重,结合灰色关联法与TOPSIS法对各项指标进行统计分析,建立三七质量评价模型,综合考察红河、文山和曲靖3个产地共15个批次的三七及其炮制品的质量。结果 检测结果表明不同产地不同批次的三七各指标水平大不相同;将三七炮制前后各项指标进行对比,发现蒸制后水分降低,浸出物含量增加,皂苷减少,多糖含量增加;15批三七及炮制品质量排序结果TOPSIS与灰色关联分析基本一致。结论 基于熵权法的灰色关联法-TOPSIS法建立的质量评价模型方便有效,可用于三七质量评价。     

不同熏硫方式对党参中二氧化硫残留量的影响

目的：探讨不同含水量、不同熏硫时间对党参中二氧化硫残留量的影响,为建立党参药材的标准化熏硫工艺研究奠定基础。方法：采用《中国药典》2010版一部附录Ⅸ U中二氧化硫残留量测定法,对熏硫后的党参中二氧化硫残留量进行测定。结果：不同水分党参经过不同时间熏硫后,发现当党参水分含量分别控制在15％、25％、35％时,熏硫时间分别控制在21h、18h、15h以内,二氧化硫低于400mg·kg^-1;水分含量在65％左右或趁鲜熏蒸后,二氧化硫残留量超过400mg·kg^-1。结论：二氧化硫残留量与熏硫时药材水分及熏硫时间有相关性,药材水分越高、熏硫时间越长,二氧化硫残留量越大。     

黄芪趁鲜切制饮片与传统饮片化学成分及体外抗氧化活性比较研究

目的 采用HPLC、UV等方法测定并比较黄芪趁鲜切制饮片与黄芪传统饮片的指标性成分及体外抗氧化活性,为不同加工方式的黄芪饮片质量控制提供科学依据。方法 采用HPLC、UV等方法对黄芪趁鲜切制饮片与黄芪传统饮片的7种指标性成分(黄芪甲苷、毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮、总黄酮、总多糖、水溶性浸出物)进行含量测定,采用熵权法结合逼近理想解排序法(technique for order preference by similarity to ideal solution,TOPSIS)评价不同产地加工方式对黄芪药材质量的影响,并结合主成分分析(principal component analysis,PCA)和偏最小二乘判别分析(partial least squares discriminant analysis,PLS-DA)等化学计量学方法对黄芪趁鲜切制饮片与黄芪传统饮片进行区分和比较,同时以DPPH自由基、羟基自由基和ABTS自由基清除能力为评价指标对黄芪趁鲜切制饮片与黄芪传统饮片的体外抗氧化活性进行比较。结果 通过熵权综合评分法比较2种不同加工方式对黄芪饮片质量的影响,结果发现黄芪趁鲜切制饮片综合评分均高于黄芪传统饮片;同时,黄芪趁鲜切制饮片组与黄芪传统饮片组在体外抗氧化活性方面,DPPH自由基清除能力的半抑制浓度(half maximal inhibitory concentration,IC₅₀)分别为5.560、8.168mg/mL,羟基自由基清除能力的IC₅₀分别为10.994、15.045mg/mL,ABTS自由基清除能力的IC₅₀分别为8.126、14.546mg/mL,表明黄芪趁鲜切制饮片体外抗氧化活性强于黄芪传统饮片。结论 通过熵权TOPSIS综合评分法结合化学计量学分析方法及体外抗氧化活性对黄芪趁鲜切制饮片与黄芪传统饮片进行分析,为不同加工方式的黄芪饮片质量控制提供参考。     

党参属种质资源多样性及可药用种质创新研究进展

桔梗科党参属Codonopsis植物全球约40种,《中国药典》2020年版仅收录党参、素花党参和川党参。党参属野生药用植物资源稀少,药材以栽培品为主。因党参需求量大,连作重茬较为普遍,种子种苗流通及全球气候变暖交融因素加速了党参种质资源性状多元化,种性混杂退化使党参根部病害加重、产量和质量降低,严重制约党参大健康产业的可持续化发展。通过文献查阅,在资源调查和田间试验基础上,对党参属植物分布、多样性、种质提纯及创新现状进行系统概述,旨在引导党参资源利用和加快新品种选育进程。     

基于化学组分特征评价不同干燥方法对麦冬品质的影响

目的通过化学组分测定和特征图谱分析,评价不同干燥方式对麦冬品质的影响。方法总皂苷、总黄酮和总多糖量采用紫外分光光度法进行测定,麦冬皂苷D和麦冬甲基黄烷酮A的量采用HPLC法进行测定;特征图谱采用HPLC法建立。结果不同干燥方法对麦冬化学组分量的影响差异显著,变异系数在6.9%~20.8%,其中,阴干、晒干、冷藏干燥和晒半干后烘干对麦冬化学组分破坏程度均较小,远红外干燥和微波干燥对麦冬化学组分破坏最大;麦冬样品有18个HPLC特征峰,聚类分析可将14种干燥方法分为3类,聚为一类的干燥方法干燥条件相近。结论通过化学组分量变化和HPLC图谱特征能够有效地反映干燥方法的差异,可作为麦冬产地干燥方法筛选的技术指标,在麦冬产地加工过程中建议晒干和空气源热泵烘干机烘干相结合应用。     

基于UPLC-Triple TOF-MS/MS技术分析不同加工方法对太子参化学成分的影响

目的 分析不同加工方法对太子参Pseudostellariae Radix中化学成分的影响。方法 采用UPLC-Triple TOF-MS/MS技术对20批样品进行测定,通过多级串联质谱分析,结合Mass数据、软件数据库及相关文献对总离子流图主要分子离子峰进行归属,数据处理用SIMCA-P软件进行多元统计分析。结果 鉴定出11种化合物;主成分得分图显示,不同加工方法太子参样品间的差异得到明显区分,通过载荷图筛选出差异显著的11种标志物,且这11种标志物呈现出不同的变化规律。结论 为揭示不同加工方法对太子参代谢产物差异性的影响规律提供科学依据。     

小柴胡颗粒制剂-药材谱峰匹配指纹图谱研究

目的 建立小柴胡颗粒制剂的多波长指纹图谱并进行药材-制剂谱峰匹配研究,进一步验证单一波长下指纹图谱质量评价的局限性。方法 采用RP-HPLC法对3个厂家各3个批次及实验室自制3批小柴胡颗粒（共12批样品）进行多波长指纹图谱相似性评价,并与相同色谱条件下的实验室自制小柴胡颗粒时所用的原药材指纹图谱进行谱峰匹配;RP-HPLC法测定不同来源的小柴胡颗粒中的指标成分的量。结果 在210、254、280、323 nm 4个波长下,同一药厂3个批次小柴胡颗粒之间及实验室自制的3批制剂之间的指纹图谱相似度差别较小,不同厂家的制剂之间,仅A厂制剂与实验室自制的3批制剂在4个波长下相似度均在0.9以上,其余二药厂除280 nm波长相似度在0.9以上外,在210、254、323 nm的相似度均小于0.9。在210、254、280、323 nm 4个波长,柴胡、甘草、党参和黄芩与小柴胡颗粒模式指纹图谱分别有5、5、2、18个色谱峰匹配;不同来源的小柴胡颗粒制剂中甘草酸和黄芩苷的量差异较大。在制剂中未能检测到柴胡皂苷a和d、阿魏酸、苍术内酯III、党参炔苷。结论 仅以280 nm波长指纹图谱相似度评价小柴胡颗粒制剂的质量存在很大的局限性。由于制剂过程及工艺的影响,部分药材的指标成分并未在制剂中测得;多波长相似度评价结合制剂用药材与制剂指纹图谱谱峰匹配及活性组分的定量测定相结合的方法,可为较全面评价中药制剂质量提供参考。