Discriminatory Components Retracing Strategy for Monitoring the Preparation Procedure of Chinese Patent Medicines by Fingerprint and Chemometric Analysis

Chinese patent medicines (CPM), generally prepared from several traditional Chinese medicines (TCMs) in accordance with specific process, are the typical delivery form of TCMs in Asia. To date, quality control of CPMs has typically focused on the evaluation of the final products using fingerprint technique and multi-components quantification, but rarely on monitoring the whole preparation process, which was considered to be more important to ensure the quality of CPMs. In this study, a novel and effective strategy labeling “retracing” way based on HPLC fingerprint and chemometric analysis was proposed with Shenkang injection (SKI) serving as an example to achieve the quality control of the whole preparation process. The chemical fingerprints were established initially and then analyzed by similarity, principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) to evaluate the quality and to explore discriminatory components. As a result, the holistic inconsistencies of ninety-three batches of SKIs were identified and five discriminatory components including emodic acid, gallic acid, caffeic acid, chrysophanol-O-glucoside, and p-coumaroyl-O-galloyl-glucose were labeled as the representative targets to explain the retracing strategy. Through analysis of the targets variation in the corresponding semi-products (ninety-three batches), intermediates (thirty-three batches), and the raw materials, successively, the origins of the discriminatory components were determined and some crucial influencing factors were proposed including the raw materials, the coextraction temperature, the sterilizing conditions, and so on. Meanwhile, a reference fingerprint was established and subsequently applied to the guidance of manufacturing. It was suggested that the production process should be standardized by taking the concentration of the discriminatory components as the diagnostic marker to ensure the stable and consistent quality for multi-batches of products. It is believed that the effective and practical strategy would play a critical role in the guidance of manufacturing and help improve the safety of the final products.     

Holistic Evaluation of Quality Consistency of Ixeris sonchifolia (Bunge) Hance Injectables by Quantitative Fingerprinting in Combination with Antioxidant Activity and Chemometric Methods

A widely used herbal medicine, Ixeris sonchifolia (Bge.) Hance Injectable (ISHI) was investigated for quality consistency. Characteristic fingerprints of 23 batches of the ISHI samples were generated at five wavelengths and evaluated by the systematic quantitative fingerprint method (SQFM) as well as simultaneous analysis of the content of seven marker compounds. Chemometric methods, i.e., support vector machine (SVM) and principal component analysis (PCA) were performed to assist in fingerprint evaluation of the ISHI samples. Qualitative classification of the ISHI samples by SVM was consistent with PCA, and in agreement with the quantitative evaluation by SQFM. In addition, the antioxidant activities of the ISHI samples were determined by both the off-line and on-line DPPH (2, 2-diphenyl-1-picryldrazyl) radical scavenging assays. A fingerprint–efficacy relationship linking the chemical components and in vitro antioxidant activity was established and validated using the partial least squares (PLS) and orthogonal projection to latent structures (OPLS) models; and the online DPPH assay further revealed those components that had position contribution to the total antioxidant activity. Therefore, the combined use of the chemometric methods, quantitative fingerprint evaluation by SQFM, and multiple marker compound analysis in conjunction with the assay of antioxidant activity provides a powerful and holistic approach to evaluate quality consistency of herbal medicines and their preparations.     

Quality control of herbal medicines

Different chromatographic and electrophoretic techniques commonly used in the instrumental inspection of herbal medicines (HM) are first comprehensively reviewed. Chemical fingerprints obtained by chromatographic and electrophoretic techniques, especially by hyphenated chromatographies, are strongly recommended for the purpose of quality control of herbal medicines, since they might represent appropriately the "chemical integrities" of the herbal medicines and therefore be used for authentication and identification of the herbal products. Based on the conception of phytoequivalence, the chromatographic fingerprints of herbal medicines could be utilized for addressing the problem of quality control of herbal medicines. Several novel chemometric methods for evaluating the fingerprints of herbal products, such as the method based on information theory, similarity estimation, chemical pattern recognition, spectral correlative chromatogram (SCC), multivariate resolution, etc. are discussed in detail with examples, which showed that the combination of chromatographic fingerprints of herbal medicines and the chemometric evaluation might be a powerful tool for quality control of herbal products.     

SVM-Based Synthetic Fingerprint Discrimination Algorithm and Quantitative Optimization Strategy

Synthetic fingerprints are a potential threat to automatic fingerprint identification systems (AFISs). In this paper, we propose an algorithm to discriminate synthetic fingerprints from real ones. First, four typical characteristic factors—the ridge distance features, global gray features, frequency feature and Harris Corner feature—are extracted. Then, a support vector machine (SVM) is used to distinguish synthetic fingerprints from real fingerprints. The experiments demonstrate that this method can achieve a recognition accuracy rate of over 98% for two discrete synthetic fingerprint databases as well as a mixed database. Furthermore, a performance factor that can evaluate the SVM''s accuracy and efficiency is presented, and a quantitative optimization strategy is established for the first time. After the optimization of our synthetic fingerprint discrimination task, the polynomial kernel with a training sample proportion of 5% is the optimized value when the minimum accuracy requirement is 95%. The radial basis function (RBF) kernel with a training sample proportion of 15% is a more suitable choice when the minimum accuracy requirement is 98%.     

Measuring What Latent Fingerprint Examiners Consider Sufficient Information for Individualization Determinations

Latent print examiners use their expertise to determine whether the information present in a comparison of two fingerprints (or palmprints) is sufficient to conclude that the prints were from the same source (individualization). When fingerprint evidence is presented in court, it is the examiner''s determination—not an objective metric—that is presented. This study was designed to ascertain the factors that explain examiners'' determinations of sufficiency for individualization. Volunteer latent print examiners (n = 170) were each assigned 22 pairs of latent and exemplar prints for examination, and annotated features, correspondence of features, and clarity. The 320 image pairs were selected specifically to control clarity and quantity of features. The predominant factor differentiating annotations associated with individualization and inconclusive determinations is the count of corresponding minutiae; other factors such as clarity provided minimal additional discriminative value. Examiners'' counts of corresponding minutiae were strongly associated with their own determinations; however, due to substantial variation of both annotations and determinations among examiners, one examiner''s annotation and determination on a given comparison is a relatively weak predictor of whether another examiner would individualize. The extensive variability in annotations also means that we must treat any individual examiner''s minutia counts as interpretations of the (unknowable) information content of the prints: saying “the prints had N corresponding minutiae marked” is not the same as “the prints had N corresponding minutiae.” More consistency in annotations, which could be achieved through standardization and training, should lead to process improvements and provide greater transparency in casework.     

Fingerprint analysis of Flos Carthami by capillary electrophoresis

Capillary electrophoresis (CE) was employed in fingerprint analysis of Flos Carthami. A standardized procedure was used to develop the CE fingerprint. An electrophoretic profile of genuine Flos Carthami from Fengqiu, He'nan, China, was first established as the characteristic fingerprint. This profile was then used to identify and assess the consistency of the herb. A study with a limited number of samples from nine sources showed a fair consistency in their CE fingerprints with that of the genuine sample. Flos Carthami was well distinguished from Stigma Croci, a possible substitute in traditional Chinese medicine, and Flos Hemerocallis, a commercial adulterant, by comparing the fingerprints of each herb.     

The Nature of Expertise in Fingerprint Matching: Experts Can Do a Lot with a Little

Expert decision making often seems impressive, even miraculous. People with genuine expertise in a particular domain can perform quickly and accurately, and with little information. In the series of experiments presented here, we manipulate the amount of “information” available to a group of experts whose job it is to identify the source of crime scene fingerprints. In Experiment 1, we reduced the amount of information available to experts by inverting fingerprint pairs and adding visual noise. There was no evidence for an inversion effect—experts were just as accurate for inverted prints as they were for upright prints—but expert performance with artificially noisy prints was impressive. In Experiment 2, we separated matching and nonmatching print pairs in time. Experts were conservative, but they were still able to discriminate pairs of fingerprints that were separated by five-seconds, even though the task was quite different from their everyday experience. In Experiment 3, we separated the print pairs further in time to test the long-term memory of experts compared to novices. Long-term recognition memory for experts and novices was the same, with both performing around chance. In Experiment 4, we presented pairs of fingerprints quickly to experts and novices in a matching task. Experts were more accurate than novices, particularly for similar nonmatching pairs, and experts were generally more accurate when they had more time. It is clear that experts can match prints accurately when there is reduced visual information, reduced opportunity for direct comparison, and reduced time to engage in deliberate reasoning. These findings suggest that non-analytic processing accounts for a substantial portion of the variance in expert fingerprint matching accuracy. Our conclusion is at odds with general wisdom in fingerprint identification practice and formal training, and at odds with the claims and explanations that are offered in court during expert testimony.     

高良姜等五味山姜属中药乙酸乙酯部位的HPLC指纹图谱

以95%乙醇作为提取的溶媒,将加热回流提取得到的提取液水浴挥干得到浸膏,并用甲醇配制成1 mg·m L~(-1)供试液。采用phenomenex-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,流速为1.0 m L·min~(-1),柱温为30℃,检测波长为260 nm HPLC色谱法测定,采用国家药典委员会相似度评价软件进行分析。结果表明:建立了精密度、稳定性和重现性均较好的5味山姜属中药乙酸乙酯部位HPLC指纹图谱,确定了9个共有峰,6个强峰;其中高良姜、大高良姜含有5个共有峰,高良姜和草豆蔻含有3个共有峰,大高良姜和红豆蔻含有2个共有峰,高良姜和益智含有3个共有峰。对5味山姜属中药乙酸乙酯部位化学成分进行相似度分析,得出其相似度分别为0.955、0.805、0.371、0.794、0.345。所建立的5味山姜属中药乙酸乙酯部位的指纹图谱方法稳定、简便、可靠,其特征峰所代表的化学成分的相似性不尽相同,高良姜、大高良姜、草豆蔻的相似性较大,红豆蔻、益智的相似性较小,说明同基原中药之间的化学成分有一定的相关性。该研究结果为探讨山姜属中药亲缘关系与化学成分相关性提供了思路与基础。     

Patent and Exclusivity Status of Essential Medicines for Non-Communicable Disease

Objective

The threat of non-communicable diseases (“NCDs”) is increasingly becoming a global health crisis and are pervasive in high, middle, and low-income populations resulting in an estimated 36 million deaths per year. There is a need to assess intellectual property rights (“IPRs”) that may impede generic production and availability and affordability to essential NCD medicines.

Methods

Using the data sources listed below, the study design systematically eliminated NCD drugs that had no patent/exclusivity provisions on API, dosage, or administration route. The first step identified essential medicines that treat certain high disease burden NCDs. A second step examined the patent and exclusivity status of active ingredient, dosage and listed route of administration using exclusion criteria outlined in this study.

Materials

We examined the patent and exclusivity status of medicines listed in the World Health Organization’s (“WHO”) Model List of Essential Drugs (Medicines) (“MLEM”) and other WHO sources for drugs treating certain NCDs. i.e., cardiovascular and respiratory disease, cancers, and diabetes. We utilized the USA Food and Drug Administration Orange Book and the USA Patent and Trademark Office databases as references given the predominant number of medicines registered in the USA.

Results

Of the 359 MLEM medicines identified, 22% (79/359) address targeted NCDs. Of these 79, only eight required in-depth patent or exclusivity assessment. Upon further review, no NCD MLEM medicines had study patent or exclusivity protection for reviewed criteria.

Conclusions

We find that ensuring availability and affordability of potential generic formulations of NCD MLEM medicines appears to be more complex than the presence of IPRs with API, dosage, or administration patent or exclusivity protection. Hence, more sophisticated analysis of NCD barriers to generic availability and affordability should be conducted in order to ensure equitable access to global populations for these essential medicines.     

Chemical fingerprint analysis of rhizomes of Gymnadenia conopsea by HPLC-DAD-MSn

A high-performance liquid chromatography-diode array detection-tandem mass spectrometry (HPLC-DAD-MS(n)) method has been firstly developed for chemical fingerprint analysis of rhizomes of Gymnadenia conopsea R. Br. and rapid identification of major compounds in the fingerprints. Comparing the UV and MS spectra with those of reference compounds, seven main peaks in the fingerprints were identified as adenosine (1), 4-hydroxybenzyl alcohol (2), 4-hydroxybenzyl aldehyde (3), dactylorhin B (4), loroglossin (5), dactylorhin A (6) and militarine (7). Compounds 4-7 were succinate derivative esters and firstly discovered from this species. The Computer Aided Similarity Evaluation System (CASES) for chromatographic fingerprint of traditional Chinese medicine was employed to evaluate the similarities of 10 samples of the rhizomes of G. conopsea collected from Sichuan, Qinghai and Hebei provinces, Tibet autonomous region of China, and Nepal. These samples from different sources had similar chemical fingerprints. This method is specific and may serve for quality identification and comprehensive evaluation of this traditional Tibetan remedy.     

Epidemiological Fingerprinting of Pseudomonas aeruginosa by the Production of and Sensitivity to Pyocin and Bacteriophage

A new method has been devised to trace cross-infection by Pseudomonas aeruginosa. Unknown strains growing logarithmically in liquid media were treated with mitomycin C to induce the liberation of pyocin and phage. The lysates were then tested against 27 selected indicator strains, and the zones of clearing were differentiated as to killing by pyocin or lysis by phage. Twenty-four standard pyocin-phage lysates were then applied to each of the unknowns, and the sensitivity pattern was recorded. Thus, an “epidemiological fingerprint” consisting of 51 operational characteristics was established for each isolate. Organisms from the same source had identical or similar fingerprints, but organisms from different origins could easily be distinguished. Pyocin production, pyocin sensitivity, and phage production were found to be stable genetic characters; however, spontaneous mutations in phage sensitivity were frequently encountered. The epidemiological fingerprint has proven to be a sensitive tool in establishing the identity or dissimilarity of unknown strains. This method has been of great value in tracing the epidemiology of P. aeruginosa in the hospital environment. Each of the 157 P. aeruginosa strains tested has been typable by this method.     

Combination with a Defucosylated Anti-HM1.24 Monoclonal Antibody plus Lenalidomide Induces Marked ADCC against Myeloma Cells and Their Progenitors

The immunomodulatory drug lenalidomide (Len) has drawn attention to potentiate antibody-dependent cellular cytotoxicity (ADCC)-mediated immunotherapies. We developed the defucosylated version (YB-AHM) of humanized monoclonal antibody against HM1.24 (CD317) overexpressed in multiple myeloma (MM) cells. In this study, we evaluated ADCC by YB-AHM and Len in combination against MM cells and their progenitors. YB-AHM was able to selectively kill via ADCC MM cells in bone marrow samples from patients with MM with low effector/target ratios, which was further enhanced by treatment with Len. Interestingly, Len also up-regulated HM1.24 expression on MM cells in an effector-dependent manner. HM1.24 was found to be highly expressed in a drug-resistant clonogenic “side population” in MM cells; and this combinatory treatment successfully reduced SP fractions in RPMI 8226 and KMS-11 cells in the presence of effector cells, and suppressed a clonogenic potential of MM cells in colony-forming assays. Collectively, the present study suggests that YB-AHM and Len in combination may become an effective therapeutic strategy in MM, warranting further study to target drug-resistant MM clonogenic cells.     

Analyzing the Distribution of Decay Constants in Pulse-Fluorimetry Using the Maximum Entropy Method

The maximum entropy method (MEM) is used to analyze time-resolved pulse-fluorescence spectrometry. The central problem in such analyses is the recovery of the distribution of exponentials describing the decay of the fluorescence (i.e., inverting the Laplace transform) which is, in turn, convolved by the shape of the excitation flash. MEM is shown to give high quality results from both computer-generated “noisy” data and experimental data from chemical and biological molecules.

The use of the Shannon-Jaynes entropy function is justified and both the theoretical and practical advantages of MEM are presented. The MEM results are easy to interpret and can help to overcome some experimental limitations. In particular MEM could be a powerful tool to analyze the heterogeneity of fluorescent emission of biological macromolecules which can be correlated with their conformational dynamics in solution.

     

How Subtle Is the “Terroir” Effect? Chemistry-Related Signatures of Two “Climats de Bourgogne”

The chemical composition of grape berries is influenced by various environmental conditions often considered to be representative of a “terroir”. If grapes from a given terroir are assumed to reflect this origin in their chemical compositions, the corresponding wine should also reflect it. The aim of this work was therefore to reveal the “terroir” expression within the chemodiversity of grapes and related wines, using ultrahigh-resolution mass spectrometry. Grapes and corresponding wines, from two distinct – though very proximate – terroirs of Burgundy were analyzed over three vintages (2010, 2011 and 2012). Ultrahigh-resolution mass spectrometry and ultra-high performance liquid chromatography were used as untargeted and targeted approaches to discriminate complex chemical fingerprints for vintages, classes (wines, skins or musts), and terroirs. Statistical analyses revealed that even if vintages have the most significant impact on fingerprints, the most significant terroir differences are seen in the grapes of a given vintage.     

Comparative Investigation for Raw and Processed Products of Euodiae Fructus Based on High-Performance Liquid Chromatography Fingerprints and Chemical Pattern Recognition

As a traditional Chinese medicine, Euodiae Fructus is widely used due to its analgesic, anti-inflammatory, and antihypertensive effects. However, Euodiae Fructus has also been documented to be toxic, and the toxic effects can be reduced by processing. To distinguish Euodiae Fructus from its processes products and study the changes of raw and processed products before and after processing, we evaluated four auxiliary material processing methods including vinegar, Zingiberis Rhizoma, Coptidis Rhizoma, and Glycyrrhizae Radix et Rhizoma. The raw Euodiae Fructus and four processed Euodiae Fructus samples were analyzed and compared based on the high-performance liquid chromatography (HPLC) fingerprints combined with chemometrics, including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and principal component analysis-class (PCA-Class). A total of 27 common peaks were obtained by fingerprint analysis. The fingerprint similarity of raw and processed samples was between 0.86–0.999. We also determined the contents of the main active ingredients - Evodiamine and Rutaecarpine. PCA and PLS-DA analyses were used to distinguish between the raw and processed samples of Euodiae Fructus, and 14 chemical markers were screened out. Four kinds of processed products were further analyzed and the results showed that they could be successfully distinguished under the established models, and 12 chemical markers were labeled. PCA-Class results revealed that the classification models constructed in this study had adequate discrimination ability. The method combined with HPLC fingerprinting and multi-component chemical pattern recognition technology could be used to differentiate raw and processed Euodiae Fructus with adequate predictive power. Our findings confirmed the rationality of the pharmacopoeial method and provided a reference for the quality control of the Glycyrrhizae Radix et Rhizoma processed Euodiae Fructus.     

Identification of the Authenticity and Geographical Origin of Bear Bile Powder by Using High Performance Liquid Chromatography - Charged Aerosol Detector Fingerprints Combined with Chemometrics

Bear bile powder (BBP) is a rare animal-derived traditional Chinese medicine, and it has been widely used to treat visual disorders and hepatobiliary diseases in East Asia. However, there is still a lack of reliable quality control methods for BBP. This study was designed to establish a comprehensive quality map of BBP based on bile acids. High-performance liquid chromatography coupled with charged aerosol detector (HPLC-CAD) was used for fingerprint establishment and quantitative analysis of BBP. The similarities of HPLC-CAD chromatograms for 50 batches of BBP were more than 0.95, while the similarities of reference chromatograms between 6 other animal bile and BBP were low than 0.7. Additionally, five bile acids in BBP, including tauroursodeoxycholic acid, taurocholic acid, taurochenodeoxycholic acid, ursodesoxycholic acid, and chenodeoxycholic acid, were simultaneously quantified. This method has been validated with good regression as well as satisfactory precision, sensitivity, stability, repeatability, and accuracy. Using this method, the contents of five bile acids in BBP samples from five producing areas were determined and compared. Furthermore, Fisher linear discriminant analysis was performed to discriminate the geographic origins of BBP. The result demonstrated that HPLC-CAD fingerprint combined with multi-components quantification is an effective and reliable method for quality control of BBP, it could be a meaningful reference for the quality evaluation of medicinal bile.     

Prediction and interpretation of the antioxidant capacity of green tea from dissimilar chromatographic fingerprints

Previously, multivariate calibration techniques have been successfully applied to model and predict the antioxidant activity of green tea from its chromatographic fingerprint. Since the selectivity differences between dissimilar chromatographic systems have already been valuably used in several applications, in this paper it is studied whether combining the complementary information contained in two dissimilar fingerprints can improve the predictive capacity of the multivariate calibration model. The simplest way of combining the data is concatenating both fingerprints for each sample. The resulting matrix can then be subjected to Orthogonal Projections to Latent Structures (O-PLS). Unfortunately, this approach resulted in a more complex model with a prediction error of about the average of the errors obtained with the individual fingerprints. Secondly, only the peaks with high loading and low orthogonal loading from both chromatograms were included in the O-PLS model. This resulted in a reduced complexity, but not in better predictions, probably due to a lack of complementarity of the information concerning the antioxidant capacity. Finally, the concatenated fingerprints were subjected to stepwise multiple linear regression (MLR) in order to build a model based on the variables most correlated with the antioxidant capacity. The obtained prediction error was lower than those of both previous approaches, but still higher than the error of the model based on a single analysis. This is probably again caused by a lack of complementarity in the variables. Nevertheless, it was advantageous to develop fingerprints on dissimilar system, because it enables to choose the most suited chromatographic profile to build a multivariate calibration model for the considered purpose. In contrast to what was expected, the study showed that the most simple (so the worst separated) fingerprints resulted in the best predictions. On the other hand, a more complex fingerprint in which more compounds are separated is still important to improve the interpretability of the model.     

滁菊的HPLC指纹图谱制作及其化学计量学分析

建立滁菊的HPLC指纹图谱,结合化学计量学手段对不同滁菊的指纹图谱进行分析研究,以期为滁菊的质量控制和产地追溯提供依据。采用HPLC法建立指纹图谱,并用中药色谱指纹图谱相似度评价系统(2012版)和系统聚类、主成分分析2种化学计量学法对指纹图谱和特征峰进行分析。分析结果发现样品有27个共有峰,12个滁菊样品具有较高的相似度,杭菊与滁菊的相似度较差,聚类分析与主成分分析结果和相似度分析结果一致。将HPLC指纹图谱与化学计量学结合可对滁菊进行鉴别和质量评价,为其质量控制和追溯提供了理论参考。     

Fingerprinting of Cyanobacteria Based on PCR with Primers Derived from Short and Long Tandemly Repeated Repetitive Sequences

The presence of repeated DNA (short tandemly repeated repetitive [STRR] and long tandemly repeated repetitive [LTRR]) sequences in the genome of cyanobacteria was used to generate a fingerprint method for symbiotic and free-living isolates. Primers corresponding to the STRR and LTRR sequences were used in the PCR, resulting in a method which generate specific fingerprints for individual isolates. The method was useful both with purified DNA and with intact cyanobacterial filaments or cells as templates for the PCR. Twenty-three Nostoc isolates from a total of 35 were symbiotic isolates from the angiosperm Gunnera species, including isolates from the same Gunnera species as well as from different species. The results show a genetic similarity among isolates from different Gunnera species as well as a genetic heterogeneity among isolates from the same Gunnera species. Isolates which have been postulated to be closely related or identical revealed similar results by the PCR method, indicating that the technique is useful for clustering of even closely related strains. The method was applied to nonheterocystus cyanobacteria from which a fingerprint pattern was obtained.     

Quality Evaluation of Potentilla fruticosa L. by High Performance Liquid Chromatography Fingerprinting Associated with Chemometric Methods

The present study was performed to assess the quality of Potentilla fruticosa L. sampled from distinct regions of China using high performance liquid chromatography (HPLC) fingerprinting coupled with a suite of chemometric methods. For this quantitative analysis, the main active phytochemical compositions and the antioxidant activity in P. fruticosa were also investigated. Considering the high percentages and antioxidant activities of phytochemicals, P. fruticosa samples from Kangding, Sichuan were selected as the most valuable raw materials. Similarity analysis (SA) of HPLC fingerprints, hierarchical cluster analysis (HCA), principle component analysis (PCA), and discriminant analysis (DA) were further employed to provide accurate classification and quality estimates of P. fruticosa. Two principal components (PCs) were collected by PCA. PC1 separated samples from Kangding, Sichuan, capturing 57.64% of the variance, whereas PC2 contributed to further separation, capturing 18.97% of the variance. Two kinds of discriminant functions with a 100% discrimination ratio were constructed. The results strongly supported the conclusion that the eight samples from different regions were clustered into three major groups, corresponding with their morphological classification, for which HPLC analysis confirmed the considerable variation in phytochemical compositions and that P. fruticosa samples from Kangding, Sichuan were of high quality. The results of SA, HCA, PCA, and DA were in agreement and performed well for the quality assessment of P. fruticosa. Consequently, HPLC fingerprinting coupled with chemometric techniques provides a highly flexible and reliable method for the quality evaluation of traditional Chinese medicines.