Variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip / 中成药

AIM To investigate the variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip.METHODS UPLC-MS/MS was adopted in the content determination of formononetin,ononin,calycosin,calycosin-7-glucoside,medicarpin,genistein,luteolin,liquiritigenin,isoliquiritigenin,vanillic acid,ferulic acid,γ-aminobutyric acid,adenosine and betaine,after which cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were used for chemical pattern recognition to explore differential components.RESULTS After rubbing strip,formononetin,calycosin,liquiritigenin and γ-aminobutynic acid demonstrated increased contents,along with decreased contents of ononin,calycosin-7-glucoside and vanillic acid.The samples with and without rubbing strip were clustered into two types,calycosin-7-glucoside,formononetin,γ-aminobutynic acid,vanillic acid,calycosin-7-glucoside and formononetin were differential components.CONCLUSION This experiment clarifies the differences of chemical constituents in Hedysari Radix before and after rubbing strip,which can provide a reference for the research on rubbing strip mechanism of other medicinal materials.

Compositional Analysis of 11 Nucleosides and Bases in Fritillaria taipaiensis P.Y.Li from Different Origins and the Differences in Their Origin / 中药新药与临床药理

Objective The contents of 11 nucleosides and base components in 10 batches of samples from 5 provinces(cities)including Chongqing,Yunnan and Shaanxi were determined,and the differences in nucleosides and base components in Fritillaria taipaiensis were compared by chemometric analysis,and the quality was comprehensively evaluated,so as to provide a reference for the cultivation of excellent varieties and the selection of medicinal materials.Methods Nucleoside and base components were extracted from Fritillaria taipaiensis by ultrasonication in aqueous solutions,and the content of each component was determined by HPLC-DAD method.The origin was classified by principal component analysis(PCA)and hierarchical cluster analysis(HCA).Partial least squares discriminant analysis(PLS-DA)was used to determine the differentiated index components in Fritillaria taipaiensis.Then the differences in the contents of the index components among samples from different origins were compared.Results It was found that 11 nucleoside and base components differed significantly among different origins of Fritillaria taipaiensis.Principal component analysis and hierarchical cluster analysis indicated that all samples could be clustered into 4 categories.Five characteristic components,including uracil,cytosine,uridine,inosine,and adenosine,were identified by PLS-DA.The nucleosides and bases in samples from Chongqing and Hubei were relatively high,and the quality of the samples was comparatively superior.Conclusion This method is simple,reproducible,accurate and reliable.It has screened out the index nucleoside and base components in the identification of Fritillaria taipaiensis of different origins,which can be used to initially elucidate the differences of samples between different origins.Additionally,it can better reflect the quality of Fritillaria taipaiensis,and can provide reference for the selection of procurement origin and the quality control for Fritillaria taipaiensis.

Study on HPLC Fingerprint and Multi-Index Components Contents of Atractylodis Macrocephalae Rhizoma / 中药新药与临床药理

Objective To establish a method for simultaneous determination of HPLC fingerprint and multi-target ingredients in Atractylodis Macrocephalae Rhizoma(AMR),in order to provide reference for its quality control.Methods HPLC-DAD multi-wavelength switching method was used to establish fingerprint of AMR,similarity evaluation combined with hierarchical clustering analysis(HCA),principal components analysis(PCA)and discriminant analysis of partial least squares(PLS-DA)were used to carry out chemometric study.The contents of differential component such as atractylenolide Ⅰ,Ⅱ,Ⅲ and atractylon were determined simultaneously.Results The HPLC fingerprint of 37 batches of AMR was established.Nine common peaks were marked,and 4 of them were identified as atractylon,atractylenolide Ⅰ,Ⅱ,Ⅲ.The similarity degrees were between 0.539 and 0.996,the quality of AMR from different origin and different batches varies greatly.Atractylon,atractylenolide Ⅰ,Ⅱ,Ⅲ and one unknown component(peak 9)are the important factors affecting the quality of AMR.Conclusion The combination methods of HPLC fingerprint and simultaneous determinations of multiple components are simple,stable,accurate and reliable,which can provide reference for the quality evaluation of AMR and the improvement of quality standard,as well as lay a foundation for the basic research of its pharmacodynamic substances and related compound.

Identification of Dalbergia odorifera and Its Counterfeits by HS-GC-MS / 中国实验方剂学杂志

ObjectiveTo screen the differential markers by analyzing volatile components in Dalbergia odorifera and its counterfeits， in order to provide reference for authentication of D. odorifera. MethodThe volatile components in D. odorifera and its counterfeits were detected by headspace gas chromatography-mass spectrometry（HS-GC-MS）， and the GC conditions were heated by procedure（the initial temperature of the column was 50 ℃， the retention time was 1 min， and then the temperature was raised to 300 ℃ at 10 ℃ for 10 min）， the carrier gas was helium， and the flow rate was 1.0 mL·min-1， the split ratio was 10∶1， and the injection volume was 1 mL. The MS conditions used electron bombardment ionization（EI） with the scanning range of m/z 35-550. The compound species were identified by database matching， the relative content of each component was calculated by the peak area normalization method， and principal component analysis（PCA）， orthogonal partial least squares-discrimination analysis（OPLS-DA） and cluster analysis were performed on the detection results by SIMCA 14.1 software， and the differential components of D. odorifera and its counterfeits were screened out according to the variable importance in the projection（VIP） value>2 and P<0.05. ResultA total of 26， 17， 8， 22， 24 and 7 volatile components were identified from D. odorifera， D. bariensis， D. latifolia， D. benthamii， D. pinnata and D. cochinchinensis， respectively. Among them， there were 11 unique volatile components of D. odorifera， 6 unique volatile components of D. bariensis， 3 unique volatile components of D. latifolia， 6 unique volatile components of D. benthamii， 8 unique volatile components of D. pinnata， 4 unique volatile components of D. cochinchinensis. The PCA results showed that， except for D. latifolia and D. cochinchinensis， which could not be clearly distinguished， D. odorifera and other counterfeits could be distributed in a certain area， respectively. The OPLS-DA results showed that D. odorifera and its five counterfeits were clustered into one group each， indicating significant differences in volatile components between D. odorifera and its counterfeits. Finally， a total of 31 differential markers of volatile components between D. odoriferae and its counterfeits were screened. ConclusionHS-GC-MS combined with SIMCA 14.1 software can systematically elucidate the volatile differential components between D. odorifera and its counterfeits， which is suitable for rapid identification of them.

Application of HPLC multi-components quantification combined with chemometrics and entropy weight-TOPSIS method in comprehensive quality evaluation of Solanum nigrum / 中国药师

Objective To establish a method for simultaneous determination of 11 components of Solanum nigrum from different producing areas,and to evaluate the quality by chemometrics and entropy weight-technique for order preference by similarity to ideal solution(EW-TOPSIS).Methods The 17 batches of Solanum nigrum samples from 8 provinces were collected.The high performance liquid chromatography(HPLC)method was used to simultaneously determine the contents of medioresino,pinoresinol,quercetin,rutoside,solasonine,solamargine,khasianine,solasodine,desgalactotigonin,diosgenin and β-sitosterol,and the multi-components quantitative control mode of Solanum nigrum was established.The quality evaluation model of Solanum nigrum was established by using chemical recognition pattern and EW-TOPSIS method,and the overall quality was evaluated comprehensively.Results When the 11 components were in the 0.78-39.00,0.55-27.50,0.34-17.00,0.21-10.50,41.87-2 093.50,60.95-3 047.50,2.58-129.00,1.02-51.00,0.46-23.00,1.05-52.50 and 0.42-21.00 μg/mL(r＞0.999 0),their linear relationships were good.The average recovery was 96.81％-100.28％with the RSD＜2.0％(n=9).17 batches of samples clustered into 3 categories.Solamargine,solasonine,desgalactotigonin and medioresino may be the main potential markers affecting the quality of Solanum nigrum.The results of EW-TOPSIS method showed that,the quality evaluation closeness of 17 batches of Solanum nigrum were 0.433 6,0.416 8,0.624 2,0.500 8,0.479 1,0.636 1,0.568 3,0.250 0,0.190 9,0.222 1,0.170 7,0.720 0,0.698 3,0.744 7,0.717 9,0.720 9 and 0.718 3,respectively,indicating that the overall quality of Solanum nigrum from Liaoning,Jilin and Heilongjiang were better,followed by Jiangsu,Henan and Anhui.Conclusion The established HPLC method for simultaneous determination of 11 components in Solanum nigrum is convenient and accurate.Chemometrics and EW-TOPSIS method are objective and comprehensive,which can be used for the overall quality evaluation of Solanum nigrum.

Study on Quality Evaluation of Classical Formula-Linggui Zhugan Decoction Based on UPLC Fingerprint Chromatogram Combined with Chemometric Analysis / 中国中医药信息杂志

Objective To establish the UPLC fingerprint chromatogram combined with chemometric analysis for the quality evaluation of classical formula Linggui Zhugan Decoction.Methods SHIMADZU Shim-Pack GIST C18 column(100 mm×2.1 mm,2.0 μm)was used with acetonitrile-0.1%phosphoric acid aqueous solution as mobile phase,gradient elution;flow rate was 0.2 mL/min;the detection wavelength was 266 nm for the first 30 minutes and 235 nm for the last 36 minutes;the column temperature was 30℃.The UPLC fingerprint of Linggui Zhugan Decoction was established by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012.130723 version),and the common peak was determined and the similarity evaluation was carried out.Based on the peak area determination results of the common peak of the fingerprint,the quality of different batches of Linggui Zhugan Decoction was evaluated by chemometrics such as clustering analysis and principal component analysis.Results A total of 24 common peaks were confirmed and 14 components were identified by using reference substances.The similarity of 10 batches of Linggui Zhugan Decoction samples was greater than 0.950,which could be divided into two categories by chemometrics,and the principal component 1-4 were the main factors affecting its quality evaluation.OPLS-DA identified 6 differential markers.Conclusion The fingerprint research method established in the study is simple,reliable and reproducible.Through the method of fingerprint combined with chemometrics analysis,the differences between Linggui Zhugan Decoction from different origins of medicinal materials are identified,which provides a reference for the internal quality evaluation of Linggui Zhugan Decoction.

DERSR-B. A Brief Scale for Detection of Emotional Dysregulation Risk / DERSR-B. Una Escala Breve para la Detección del Riesgo de Desregulación Emocional

Abstract Emotional dysregulation (ED) is related to problems in understanding, perceiving, and regulating emotions. The aim is to find the psychometric properties of an instrument that measures ED and classifies the high/low ED group membership with the least possible error. For statistical purposes (factor analysis), two independent samples of males and females (n1 = 476) and (n2 = 562) were obtained, with ages sample 1 (15 -19 years; M= 15.8; SD=0.71) and sample 2 (15-19 years; M=15.6; SD= 0.69). Three factors were formed by sex, males with 14 items and females with 13 items, each loading on a single factor (total α=0.71 - 0.78 ɷ =0.67- 0.79 females; α= 0.70 - 0.79 ɷ=0.73 - 0.75 males) and good fit indices. In sum, a validated cut version instrument (DERSR-B), a risk screening instrument, was obtained.

Resumen La desregulación emocional (DE) se relaciona con problemas para comprender, percibir y regular las emociones. Determinar las propiedades psicométricas de un instrumento que mide DE y que clasifica con el menor error posible la pertenencia de grupo alto/bajo de DE se propuso como el objetivo de este estudio. Para propósitos estadísticos (análisis factoriales) se obtuvieron dos muestras independientes de hombres y mujeres (n1 = 476) y (n2 = 562) respectivamente, con edades para muestra 1 (15-19 años; M= 15.8; DE= 0.71) y muestra 2 (15-19 años; M=15.6; DE= 0.69). Se obtuvieron tres factores por sexo, hombres con 14 ítems y mujeres con 13 ítems cada uno cargando en un solo, un único factor (total α =0.71 - 0.78 ɷ =0.67- 0.79 mujeres; α=0.70 - 0.79 ɷ=0.73-0.75 hombres) y con índices de ajuste aceptables. Se obtuvo un instrumento válido en versión corta de detección rápida de riesgo de Desregulación Emocional (DERSR-B).

Effects of Abacus Training on Auditory Spatial Maturation in Children with Normal Hearing

Abstract Introduction The spatial auditory system, though developed at birth, attains functional maturity in the late childhood (12 years). Spatial changes during childhood affect navigation in the environment and source segregation. Accommodation of a new skill through learning, especially during childhood, can expedite this process. Objective To explore the auditory spatial benefits of abacus training on psychoacoustic metrics in children. The study also aimed to identify the most sensitive metric to abacus training related changes in spatial processing, and utilize this metric for a detailed spatial error profiling. Methods A standard group comparison analysis with 90 participants divided into three groups: I: children with abacus training (C-AT); II: children with no training (C-UT); III: adults with no training (A-UT). The groups underwent a series of psychoacoustic tests, such as interaural time difference (ITD), interaural level difference (ILD), and virtual auditory space identification (VASI), as well as perceptual tests such as the Kannada version of the speech, spatial, and quality questionnaire (K-SSQ). Results Significant group differences were observed in the multivariate analysis of variance (MANOVA) and post-hoc tests, with the C-AT group showing significantly lower ILD scores (p = 0.01) and significantly higher VASI scores (p < 0.001) compared to the CUT group, which is indicative of better spatial processing abilities in the former group. The discriminant function (DF) analyses showed that the VASI was the most sensitive metric for training-related changes, based on which elaborate error analyses were performed. Conclusions Despite the physiological limits of the immature neural framework, the performance of the C-AT group was equivalent to that of untrained adults on psychoacoustic tests, which is reflective of the positive role of abacus training in expediting auditory spatial maturation.

Discrimination of cultivation modes of Dendrobium nobile based on content of mineral elements and ratios of nitrogen stable isotopes / 中国中药杂志

This study explored the feasibility of mineral element content and ratios of nitrogen isotopes to discriminate the cultivation mode of Dendrobium nobile in order to provide theoretical support for the discrimination of the cultivation mode of D. nobile. The content of 11 mineral elements(N, K, Ca, P, Mg, Na, Fe, Cu, Zn, Mn, and B) and nitrogen isotope ratios in D. nobile and its substrate samples in three cultivation methods(greenhouse cultivation, tree-attached cultivation, and stone-attached cultivation) were determined. According to the analysis of variance, principal component analysis, and stepwise discriminant analysis, the samples of different cultivation types were classified. The results showed that the nitrogen isotope ratios and the content of elements except for Zn were significantly different among different cultivation types of D. nobile(P<0.05). The results of correlation analysis showed that the nitrogen isotope ratios, mineral element content, and effective component content in D. nobile were correlated with the nitrogen isotope ratio and mineral element content in the corresponding substrate samples to varying degrees. Principal component analysis can preliminarily classify the samples of D. nobile, but some samples overlapped. Through stepwise discriminant analysis, six indicators, including δ~(15)N, K, Cu, P, Na, and Ca, were screened out, which could be used to establish the discriminant model of D. nobile cultivation methods, and the overall correct discrimination rates after back-substitution test, cross-check, and external validation were all 100%. Therefore, nitrogen isotope ratios and mineral element fingerprints combined with multivariate statistical analysis could effectively discriminate the cultivation types of D. nobile. The results of this study provide a new method for the identification of the cultivation type and production area of D. nobile and an experimental basis for the quality evaluation and quality control of D. nobile.

Simultaneous determination of eleven volatile components in Cinnamomi Oleum by GC-MS / 中国中药杂志

A gas chromatography-triple quadrupole mass spectrometry(GC-MS) method was established for the simultaneous determination of eleven volatile components in Cinnamomi Oleum and the chemical pattern recognition was utilized to evaluate the quality of essential oil obtained from Cinnamomi Fructus medicinal materials in various habitats. The Cinnamomi Fructus medicinal materials were treated by water distillation, analyzed using GC-MS, and detected by selective ion monitoring(SIM), and the internal standards were used for quantification. The content results of Cinnamomi Oleum from various batches were analyzed by hierarchical clustering analysis(HCA), principal component analysis(PCA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) for the statistic analysis. Eleven components showed good linear relationships within their respective concentration ranges(R~2>0.999 7), with average recoveries of 92.41%-102.1% and RSD of 1.2%-3.2%(n=6). The samples were classified into three categories by HCA and PCA, and 2-nonanone was screened as a marker of variability between batches in combination with OPLS-DA. This method is specific, sensitive, simple, and accurate, and the screened components can be utilized as a basis for the quality control of Cinnamomi Oleum.

Metabonomics Analysis of Brain Stem Tissue in Rats with Primary Brain Stem Injury Caused Death / 法医学杂志

OBJECTIVES@#To explore the potential biomarkers for the diagnosis of primary brain stem injury (PBSI) by using metabonomics method to observe the changes of metabolites in rats with PBSI caused death.@*METHODS@#PBSI, non-brain stem brain injury and decapitation rat models were established, and metabolic maps of brain stem were obtained by LC-MS metabonomics method and annotated to the HMDB database. Partial least square-discriminant analysis (PLS-DA) and random forest methods were used to screen potential biomarkers associated with PBSI diagnosis.@*RESULTS@#Eighty-six potential metabolic markers associated with PBSI were screened by PLS-DA. They were modeled and predicted by random forest algorithm with an accuracy rate of 83.3%. The 818 metabolic markers annotated to HMDB database were used for random forest modeling and prediction, and the accuracy rate was 88.9%. According to the importance in the identification of cause of death, the most important metabolic markers that were significantly up-regulated in PBSI group were HMDB0038126 (genipinic acid, GA), HMDB0013272 (N-lauroylglycine), HMDB0005199 [(R)-salsolinol] and HMDB0013645 (N,N-dimethylsphingosine).@*CONCLUSIONS@#GA, N-lauroylglycine, (R)-salsolinol and N,N-dimethylsphingosine are expected to be important metabolite indicators in the diagnosis of PBSI caused death, thus providing clues for forensic medicine practice.

Forensic Pathological Diagnosis of Acute and Old Myocardial Infarction Using Fourier Transform Infrared Spectroscopy / 法医学杂志

OBJECTIVES@#Fourier transform infrared spectroscopy (FTIR) was used to analyze myocardial infarction tissues at different stages of pathological change to achieve the forensic pathology diagnosis of acute and old myocardial infarction.@*METHODS@#FTIR spectra data of early ischemic myocardium, necrotic myocardium, and myocardial fibrous tissue in the left ventricular anterior wall of the sudden death group of atherosclerotic heart disease and the myocardium of the normal control group were collected using hematoxylin-eosin (HE) and immunohistochemistry (IHC) staining as a reference, and the data were analyzed using multivariate statistical analysis.@*RESULTS@#The mean normalized spectra of control myocardium, early ischemic myocardium and necrotic myocardium were relatively similar, but the mean second derivative spectra were significantly different. The peak intensity of secondary structure of proteins in early ischemic myocardium was significantly higher than in other types of myocardium, and the peak intensity of the α-helix in necrotic myocardium was the lowest. The peaks of amide Ⅰ and amide Ⅱ in the mean normalized spectra of myocardial fibrous tissue significantly shifted towards higher wave numbers, the peak intensities of amide Ⅱ and amide Ⅲ were higher than those of other types of myocardium, and the peak intensities at 1 338, 1 284, 1 238 and 1 204 cm-1 in the mean second derivative spectra were significantly enhanced. Principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) showed that FTIR could distinguish different types of myocardium.@*CONCLUSIONS@#FTIR technique has the potential to diagnose acute and old myocardial infarction, and provides a new basis for the analysis of the causes of sudden cardiac death.

Characteristics of the urinary microbiota in UTUC patients / 中华泌尿外科杂志

Objective:To analyzed the urinary microbiota characteristics of upper tract urothelial carcinoma(UTUC) patients.Methods:Urine samples were collected from 23 patients with UTUC (UTUC group) and 22 patients with benign diseases (control group) admitted to Yijishan Hospital, the First Affiliated Hospital of Wannan Medical College from July 2021 to July 2022. The differences in age [(60.9±5.7) years vs. (61.4±8.8) years], sex (male/female: 15/8 vs. 9/13), and body mass index [(22.9±1.8) kg/m 2 vs. (23.4±1.7) kg/m 2] between the UTUC group and the control group were not statistically significant ( P>0.05). The V4 region of the 16S rRNA of urinary microorganisms was sequenced using the Illumina NovaSeq6000, and the results were processed using QLLME2. Differences in α-diversity between groups were analyzed by using the Shannon, Simpson, and Chao1 indices. Differences in β-diversity between groups were analyzed by using unweighted principal coordinates analysis (PCoA). Linear discriminant analysis Effect Size(LEfSe)was used to identify the bacterial taxa with different abundances between groups. Significant differences were defined as LDA>2. Results:The Chao1 index (703.12±265.54 vs. 506.20±214.02) and Shannon index (5.61±1.85 vs. 5.07±1.34) were significantly higher in the UTUC group compared to that in the control group ( P<0.05). The α-diversity of urinary microbes was elevated in the UTUC group compared to that in the control group but the difference in β-diversity was not statistically significant ( P=0.161). The enrichment of Bacteroidaceae, Ruminococcaceae, Acidaminococcaceae, Thermaceae, Erysipelatoclostridiaceae, and Coriobacteriaceae abundance was higher in the urine of UTUC patients(LDA > 2). Further subgrouping analyses of the UTUC patients showed that the differences in Chao1 index (706.44±271.84 vs. 784.09±272.72), Shannon index (6.04±1.30 vs. 5.91±1.67), and Simpson index (0.94±0.08 vs. 0.89±0.22) between the muscle-invasive group and the non-muscle-invasive group were not statistically significant ( P>0.05). The difference in α-diversity between muscle-invasive and non-muscle-invasive group was not statistically significant, but the difference in β-diversity was statistically significant ( P=0.047). The urinary microbial communities of Gammaproteobacteria, Cutibacterium, Rhodococcus and Nocardiaceae were enriched in muscle-invasive group and differed from that in non-muscle-invasive group(LDA>2). Conclusions:This study suggests that the urinary microbial community was more abundant in UTUC patients than in non-UTUC patients and that Bacteroidaceae, Ruminococcaceae, Acidaminococcaceae, Thermaceae, Erysipelatoclostridiaceae, and Coriobacteriaceae were more abundant in the urine of UTUC patients. The urinary microbial community was more abundant in the urine of non-muscle-invasive patients than in the muscle-invasive patients, and Gammaproteobacteria, Cutibacterium, Rhodococcus and Nocardiaceae were more abundant in the urine of non-muscle-invasive patients.

UHPLC-Q-Orbitrap/MS-based analysis of saponins from Panacis Quinquefolii Radix of different origins / 中成药

AIM To analyze saponins from Panacis Quinquefolii Radix of different regions(Jilin,Liaoning,Heilongjiang,Shandong)based on UHPLC-Q-Orbitrap/MS technology.METHODS Panacis Quinquefolii Radix had its saponins qualitatively analyzed by UHPLC-Q-Orbitrap/MS;and its differential saponins revealing different regions screened by the principal component analysis,orthogonal partial least squares discriminant analysis and differential component analysis.RESULTS A total of 62 saponins were identified;saponin variations due to the growth areas verified by the principal component analysis;and 28 differential saponins including 13 protopanaxadiol,6 protopanaxatriol,4 oleanolane,2 oxytetracycline,and 3 C-17 side chain variants further screened by orthogonal partial least-squares discriminant analysis model.The relative contents of protopanaxadiol-type and protopanaxatriol-type saponins in the four producing areas reduced following the order of Liaoning,Jilin,Heilongjiang,and Shandong.CONCLUSION This simple,accurate and efficient method provides a theoretical basis for the quality evaluation of Panacis Quinquefolii Radix.

Study on HPLC Fingerprint of Alpiniae Oxyphyllae Fructus Before and After Salt-Processing Based on Chemical Pattern Recognition / 中药新药与临床药理

Objective A HPLC fingerprint method of Alpiniae Oxyphyllae Fructus(AOF)before and after salt-processing was established,to compare the differences of chemical components between raw and processed AOF combined with chemical pattern recognition.Methods HPLC method was used to establish the fingerprint of raw and salt-processed AOF.Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were applied to explore the different components of raw and salt-processed AOF in different batches.Results Totally 30 and 32 common peaks in the HPLC fingerprint from the raw and salt-processed AOF were detected,respectively.And 8 of them were identified by comparison with the standards.They were peak X2(5-hydroxymethylfurfural),peak 1(protocatechuic acid),peak 2(protocatechualdehyde),peak 4(epicatechin),peak 21(chrysin),peak 22(kaempferide),peak 25(tectochrysin)and peak 26(nootkatone).The results of PCA and OPLS-DA showed that raw and salt-processed AOF can be grouped into two categories.A total of 12 components,which were considered as differential markers of raw and salt-processed AOF,were screened by method of variable importance in projection(VIP).The 12 components were peak X1,peak 26(nootkatone),peak 16,peak 3,peak X2(5-hydroxymethylfurfural),peak 25(tectochrysin),peak 15,peak 12,peak 8,peak 10,peak 17 and peak 20.Conclusion The combination of HPLC fingerprint and chemical pattern recognition can be used to analyze the quality differences of AOF before and after salt-processing.

Comparison of Effect of Ancient and Modern Processing Methods on Chemical Components of Lilii Bulbus Decoction Based on UHPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

ObjectiveTo compare the effects of different processing methods in ancient and modern times on the chemical components of Lilii Bulbus decoction， and to provide experimental support for the origin processing， decoction piece processing and clinical application of this herb. MethodUltra high performance liquid chromatography tandem quadrupole electrostatic field orbitrap high resolution mass spectrometry（UHPLC-Q-Orbitrap HRMS） was used for structural identification of the compounds using excimer ions， secondary MS and characteristic fragment ions， and referring to relevant literature and database information. Principal component analysis（PCA） and orthogonal partial least squares discriminant analysis（OPLS-DA） were used to screen the main differential components， the differential components were quantitatively studied by high performance liquid chromatography（HPLC）， in order to compare the types and contents of chemical components in the decoction of different processing products of Lilii Bulbus. ResultA total of 24 chemical components were identified from the decoction of different processed products of Lilii Bulbus， water extract and scalding liquid of fresh Lilii Bulbus， including 17 phenols， 5 saponins and 2 alkaloids. Compared with the fresh Lilii Bulbus decoction， the contents of regaloside A， p-coumaric acid， colchicine and other components in the decoction of dry Lilii Bulbus processed by scalding method decreased， the content of regaloside C in the decoction of dry Lilii Bulbus processed by steaming method decreased， and the contents of regaloside A and regaloside C in the decoction of fresh Lilii Bulbus processed by water immersion also decreased. Compared with the decoction of dry Lilii Bulbus processed by scalding method， the overall content of components in the fresh Lilii Bulbus decoction and the decoction of fresh Lilii Bulbus processed by water immersion was higher， the contents of components in the decoction of dry Lilii Bulbus processed by steaming method was higher， except for the slightly lower content of regaloside C. ConclusionDifferent processing processes have a certain effect on the types and contents of chemical components in Lilii Bulbus decoction. Scalding process is beneficial to the preservation of Lilii Bulbus， but can cause the loss of effective components. Compared with scalding method， steaming method can prevent browning of Lilii Bulbus and reduce the loss of its active ingredients. The processing method of removing foam after overnight immersion proposed by ZHANG Zhongjing may be more conducive to the treatment of Baihe disease， which can provide reference for the clinical rational application and mechanism research of different processed products of Lilii Bulbus.

HPLC Fingerprint Analysis of Benchmark Sample of Yanghetang / 中国实验方剂学杂志

ObjectiveTo establish a high performance liquid chromatography（HPLC） fingerprint of Yanghetang benchmark sample， and evaluate its quality with chemometric methods， so as to provide a reference for the quality control of this benchmark sample. MethodHPLC was used to establish the fingerprint of Yanghetang benchmark sample with ZORBAX SB-C18 column（4.6 mm×250 mm， 5 μm）， the mobile phase was consisted of acetonitrile（A） -0.05% phosphoric acid aqueous solution （containing 0.05% triethylamine solution）（B） for gradient elution（0-5 min， 2%-3%A； 5-15 min， 3%-5%A； 15-65 min， 5%-30%A； 65-90 min， 30%-70%A）， the flow rate was 1.0 mL·min-1， the column temperature was 35 ℃， and the detection wavelength was 210， 260 nm. Traditional Chinese Medicine（TCM） Chromatographic Fingerprint Similarity Evaluation System （2012 edition） combined with cluster analysis， principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA） were used to evaluate the quality differences between different batches of Yanghetang benchmark samples， and to find the main chemical components responsible for the quality differences. ResultHPLC fingerprint of Yanghetang benchmark sample was established， 13 common peaks were identified and attributed to each common peak， including peaks 2 and 8 from Rehmanniae Radix Praeparata， peaks 10 and 11 from Cinnamomi Cortex， peaks 1， 3-6 from fried Sinapis Semen， peak 13 from Ephedrae Herba， and peaks 7， 9， 12 from Glycyrrhizae Radix et Rhizoma. Eight of them were identified by comparing with control substance， which were 5-hydroxymethylfurfural（peak 2）， sinapine thiocyanate（peak 4）， glycyrrhizin（peak 7）， verbascoside（peak 8）， cinnamic acid（peak 10）， cinnamaldehyde（peak 11）， glycyrrhizic acid（peak 12） and ephedrine hydrochloride（peak 13）. The similarities of the HPLC fingerprints of 15 batches of Yanghetang benchmark samples with the control fingerprint were all greater than 0.80. The three chemometric methods could classify the samples into two categories. Eight differential components were screened out， among which 5-hydroxymethylfurfural， sinapine thiocyanate， verbascoside and ephedrine hydrochloride were identified. ConclusionThe established fingerprint analysis method is accurate， stable and reproducible， which basically reflects the overall chemical composition of Yanghetang benchmark sample， and can provide a basis for establishment of quality standards for compound preparations of this famous classical formula.

Comparison on volatile components between Artemisiae Verlotori Folium and Artemisiae Argyi Folium based on GC-MS and chemometrics / 中国中药杂志

Artemisiae Argyi Folium is commonly used in clinical practice. Artemisiae Verlotori Folium, the dried leaves of Artemisia verlotorum, is often used as a folk substitute for Artemisiae Argyi Folium in Lingnan area. In this study, gas chromatography-triple quadrupole mass spectrometry(GC-MS) was used to detect the volatile oil components of 27 samples of Artemisiae Verlotori Folium and 13 samples of Artemisiae Argyi Folium, and the volatile components were compared between the two species. The internal standard method was combined with multi-reaction monitoring mode(MRM) to determine the content of six major volatile components. Hierarchical clustering analysis(HCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were carried out for the content data. The results showed that the Artemisiae Argyi Folium samples had higher content and more abundant volatile oils than the Artemisiae Verlotori Folium samples. Artemisiae Argyi Folium mainly had the components with lower boiling points, while Artemisiae Verlotori Folium mainly had the components with higher boiling points. Terpenoids were the main volatile components in Artemisiae Verlotori Folium(mainly sesquiterpenoids) and Artemisiae Argyi Folium(monoterpenoids). In addition, Artemisiae Argyi Folium had higher content of oxygen-containing derivatives than Artemisiae Verlotori Folium. Furthermore, the stoichiometric analysis showed that the two species could be distinguished by both HCA and OPLS-DA, indicating that the volatile components of the two were significantly different. This study can provide a scientific basis for the quality evaluation and data support for the local rational application of Artemisiae Verlotori Folium in Lingnan.

Postmortem Interval Estimation Using Protein Chip Technology Combined with Multivariate Analysis Methods / 法医学杂志

OBJECTIVES@#To estimate postmortem interval (PMI) by analyzing the protein changes in skeletal muscle tissues with the protein chip technology combined with multivariate analysis methods.@*METHODS@#Rats were sacrificed for cervical dislocation and placed at 16 ℃. Water-soluble proteins in skeletal muscles were extracted at 10 time points (0 d, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, 7 d, 8 d and 9 d) after death. Protein expression profile data with relative molecular mass of 14 000-230 000 were obtained. Principal component analysis (PCA) and orthogonal partial least squares (OPLS) were used for data analysis. Fisher discriminant model and back propagation (BP) neural network model were constructed to classify and preliminarily estimate the PMI. In addition, the protein expression profiles data of human skeletal muscles at different time points after death were collected, and the relationship between them and PMI was analyzed by heat map and cluster analysis.@*RESULTS@#The protein peak of rat skeletal muscle changed with PMI. The result of PCA combined with OPLS discriminant analysis showed statistical significance in groups with different time points (P<0.05) except 6 d, 7 d and 8 d after death. By Fisher discriminant analysis, the accuracy of internal cross-validation was 71.4% and the accuracy of external validation was 66.7%. The BP neural network model classification and preliminary estimation results showed the accuracy of internal cross-validation was 98.2%, and the accuracy of external validation was 95.8%. There was a significant difference in protein expression between 4 d and 25 h after death by the cluster analysis of the human skeletal muscle samples.@*CONCLUSIONS@#The protein chip technology can quickly, accurately and repeatedly obtain water-soluble protein expression profiles in rats' and human skeletal muscles with the relative molecular mass of 14 000-230 000 at different time points postmortem. The establishment of multiple PMI estimation models based on multivariate analysis can provide a new idea and method for PMI estimation.

Comprehensive quality evaluation of Patrina scabiosaefolia Fisch based on HPLC multi-component quantitative control and PCA,OPLS-DA combined with gray correlation analysis / 中国药师

Objective To establish a HPLC method for the simultaneous determination of hederagenin,oleanolic acid,ursolic acid,scabioside C,protocatechuic acid,chlorogenic acid,caffeic acid,aesculetin,scopoletin,rutoside,quercetin,kaempferol in Patrina scabiosaefolia Fisch,and to explore the application of principal component analysis(PCA),orthogonal partial least squares-discriminant analysis(OPLS-DA)and grey relational analysis(GRA)in the quality evaluation of Patrina scabiosaefolia Fisch.Methods The HPLC method was used with the Waters Atlantis T3 C18 column(250 mm×4.6 mm,5 μm).The mobile phase was acetonitrile-0.1% phosphate acid solution with gradient elution.The detection wavelengths were 210,260 and 360 nm.The overall quality of Patrina scabiosaefolia Fisch was comprehensively evaluated by PCA,OPLS-DA and GRA analysis of the results of multi-component content.Results The results of methodological verification of external standard method met the requirements.The 12 components had good linear relationships within their respective ranges(r>0.999 0),and the average recovery rate were 96.82% -100.16% (RSD<2.0% ,n=9).PCA and OPLS-DA results showed that oleanolic acid,ursolic acid,chlorogenic acid and quercetin were the main potential markers affecting the quality of Patrina scabiosaefolia Fisch.The relative correlation degree of GRA were 0.380 6-0.571 4,and there was a certain difference between batches of Patrina scabiosaefolia Fisch.Conclusion The HPLC method can simultaneously determine the 12 components in Patrina scabiosaefolia Fisch,which is simple and practical,and PCA,OPLS-DA and GRA can be used to evaluate the quality of Patrina scabiosaefolia Fisch.