1.Characteristic ion Identification of Different Original Haliotidis Concha and Its Counterfeits
Xiaojie LIANG ; Guowei LI ; Lin ZHOU ; Qiping HU ; Muxiang LUO ; Jiehao TANG ; Xiangdong CHEN ; Liye PAN ; Dongmei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):263-269
ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits, and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai, 4 batches of H. ruber, 3 batches of H. laevigata, 3 batches of H. ovina, 3 batches of H. diversicolor, 3 batches of H. asinina, 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive-Orbitrap-MS/MS) was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits, and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry(UPLC-QqQ-MS/MS) was used to validate the characteristic ions, and the specific detection method of the characteristic ions was established. ResultsA total of 1 182, 167, 47, 89, 104, 203, 424 potential characteristic ions were screened from H. discus hannai, H. ruber, H. laevigata, H. ovina, H. diversicolor, H. asinina and H. iris, respectively. And 9 characteristic ions were selected. The precision, stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions, including m/z 631.83-886.48(double charge) and m/z 631.83-443.74(double charge) of H. discus hannai, m/z 699.28-232.11(double charge) and m/z 699.28-544.27(double charge) of H. ruber, m/z 535.76-752.37(double charge) and m/z 535.76-548.28(double charge) of H. laevigata, m/z 708.35-442.28(double charge) and m/z 708.35-215.14(double charge) of H. ovina, m/z 561.33-614.86(triple charge), m/z 561.33-468.28(triple charge), m/z 608.29-618.32(double charge) and m/z 608.29-390.21(double charge) of H. diversicolor, m/z 769.85-274.10(double charge), m/z 769.85-532.75(double charge), m/z 827.43-646.36(single charge), m/z 827.43-257.12(single charge) of H. asinina, and m/z 468.24-576.29(double charge) and m/z 468.24-505.26(double charge) of H. iris. ConclusionIn this study, a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits, and a specific identification method is established, which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha, and provide a basis for the production, circulation and medication quality.
2.Characteristic ion Identification of Different Original Haliotidis Concha and Its Counterfeits
Xiaojie LIANG ; Guowei LI ; Lin ZHOU ; Qiping HU ; Muxiang LUO ; Jiehao TANG ; Xiangdong CHEN ; Liye PAN ; Dongmei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):263-269
ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits, and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai, 4 batches of H. ruber, 3 batches of H. laevigata, 3 batches of H. ovina, 3 batches of H. diversicolor, 3 batches of H. asinina, 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive-Orbitrap-MS/MS) was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits, and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry(UPLC-QqQ-MS/MS) was used to validate the characteristic ions, and the specific detection method of the characteristic ions was established. ResultsA total of 1 182, 167, 47, 89, 104, 203, 424 potential characteristic ions were screened from H. discus hannai, H. ruber, H. laevigata, H. ovina, H. diversicolor, H. asinina and H. iris, respectively. And 9 characteristic ions were selected. The precision, stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions, including m/z 631.83-886.48(double charge) and m/z 631.83-443.74(double charge) of H. discus hannai, m/z 699.28-232.11(double charge) and m/z 699.28-544.27(double charge) of H. ruber, m/z 535.76-752.37(double charge) and m/z 535.76-548.28(double charge) of H. laevigata, m/z 708.35-442.28(double charge) and m/z 708.35-215.14(double charge) of H. ovina, m/z 561.33-614.86(triple charge), m/z 561.33-468.28(triple charge), m/z 608.29-618.32(double charge) and m/z 608.29-390.21(double charge) of H. diversicolor, m/z 769.85-274.10(double charge), m/z 769.85-532.75(double charge), m/z 827.43-646.36(single charge), m/z 827.43-257.12(single charge) of H. asinina, and m/z 468.24-576.29(double charge) and m/z 468.24-505.26(double charge) of H. iris. ConclusionIn this study, a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits, and a specific identification method is established, which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha, and provide a basis for the production, circulation and medication quality.
3.Study on the changes of chemical components of Aristolochia contorta before and after honey processing based on HPLC fingerprint and multivariate statistical analysis
Xiaoxia LIU ; Yongwei FENG ; Cuijie WEI ; Chunxiu WU ; Minyou HE ; Liye PAN ; Dongmei SUN ; Zhenyu LI
International Journal of Traditional Chinese Medicine 2024;46(11):1485-1492
Objective:To establish HPLC fingerprints of Aristolochia contorta and honey-processed Aristolochia contorta; To analyze the changes of chemical components before and after honey processing with multivariate statistics; To provide a reference for the study on the toxicity reduction of Aristolochia contorta.Methods:The fingerprints of 11 batches of Aristolochia contorta and honey-processed Aristolochia contorta were established through HPLC. Clustering analysis (HCA), principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA) and independent sample t-test were used to compare the changes of chemical components of Aristolochia contorta before and after honey processing.Results:The results showed that there were 14 common peaks in the fingerprints of Aristolochia contorta and Aristolochia contorta. 7 common peaks were identified. Both HCA and PCA could clearly distinguish the samples of Aristolochia contorta before and after honey processing. OPLS-DA found and screened 7 differential markers, and the order of difference significance was peak 3 > peak 7 (7-hydroxy aristolochic acid A) > peak 5 (aristolochic acid C)> peak 8 (aristolochic acid D) > peak 6 > peak 2 (Magnolia alkaloid) > peak 14 (aristolochic acid Ⅰ). After honey processing, the content of chemical components represented by peaks 2, 3, 5, 6, 7, 8 and 14 decreased ( P<0.05). Conclusion:This method is simple and specific, which can be used for the fingerprint analysis of Aristolochia contorta and honey-processed Aristolochia contorta, and can effectively distinguish Aristolochia contorta and honey-processed Aristolochia contorta, and provide a reference for the processing research of toxicity reduction of Aristolochia contorta honey processing.
4.Quality study of Prunellae spica dispensing granules based on standard decoction
Zhenyu LI ; Qing DING ; Wanfa CHEN ; Liwei WANG ; Yueyi LIANG ; Xiaojun PAN ; Xiangyuan ZHOU ; Liye PAN ; Wenjie HUO
International Journal of Traditional Chinese Medicine 2022;44(2):188-194
Objective:To establish the quality evaluation method of Prunellae spica dispensing granules based on three quality indexes of standard decoction. Methods:Fourteen batches of Prunellae spica were collected from different habitats. According to technical requirements, fourteen batches of Prunellae spica standard decoction and three batches of formula granules were prepared and the paste-forming rates were calculated. The fingerprints of Prunellae spica standard decoction and formula granules were established by Ultra High Performance Liquid Chromatography (UPLC). The similarity values of fingerprints between dispensing granules and standard decoction were calculated. The content and transferring rate of Rosmarinic acid were determined and calculated. Results:The average paste-forming rate of Prunellae spica was (12.59±2.32)%. The paste-forming rates of the three batches were 11.14%, 10.78% and 10.39% respectively. The average content of Rosmarinic acid in standard decoction was (18.99±9.74)mg/g. The average transferring rate was (60.58±7.87)%. The contents of three batches were 7.40 mg/g, 7.49 mg/g and 7.09 mg/g. The transferring rates were 52.06%, 50.10% and 50.40% respectively. Nine common fingerprint peaks were identified in the fingerprints of standard decoction and formula granules, two of which were identified as Rosmarinic acid and Caffeic acid by comparison of reference substance. The fingerprints similarity of Prunellae spica dispensing granules and standard decoction were 0.954, 0.973 and 0.952, respectively. Conclusions:The quality indexes of three batches of formulation granules are consistent with standard decoction. This method could provide reference for the establishment of quality standard of Prunellae spica dispensing granules.
5.Establishment of UPLC Fingerprint of Pyrrosia petiolosa from Southwest China and Content Determination of Phenolic Acids Component
Yunjing QIU ; Caixian SUO ; Liye PAN ; Minyou HE ; Xiangdong CHEN ; Guowei LI
China Pharmacy 2021;32(9):1093-1099
OBJECTIVE:To esta blish a UPLC fingerprint of Pyrrosia petiolosa from southwest China ,and to determine the contents of 4 kinds of phenolic acids (neochlorogenic acid ,caffeic acid ,chlorogenic acid and cryptochlorogenic acid ). METHODS:The determination was performed on Waters Cortecs T 3 C18 column(100 mm×2.1 mm,1.6 μm)with mobile phase consisted of methanol- 0.1% phosphoric acid (gradient elution )at the flow rate of 0.35 mL/min. The detection wavelength was set at 326 nm. The column temperature was 30 ℃,and injection volume was 1 μL. UPLC method was used to establish the UPLC fingerprint of P. petiolosa in combination with the Similarity Evaluation System of TCM Chromatographic Fingerprints (2012 edition). Cluster analysis and principle component analysis (PCA)were performed by using SPSS 20.0 software. The contents of 4 kinds of phenolic acids in 20 batches of P. petiolosa were determined by external standard method. RESULTS :There were 9 common peaks for the UPLC fingerprint of P. petiolosa . Peaks 1,3,4,5 and 9 were identified as neochlorogenic acid ,caffeic acid,chlorogenic acid ,cryptochlorogenic acid and isochlorogenic acid C ,respectively. RSDs of the relative retention time of each peak in different batches of P. petiolosa were 0-0.68%,and the RSDs of the relative peak area were 0-62.35%. The similarities between the fingerprint of 20 batches of medicinal materials and the control chromatogram were not less than 0.990. The result of cluster analysis showed that P. petiolosa from different regions could be sorted into three species. Results of PCA showed the differences among P. petiolosa from different regions. The linear range of neochlorogenic acid ,caffeic acid ,chlorogenic acid and cryptochlorogenic acid were 0.61-61.41,0.18-17.60,2.00-200.11,0.62-61.51 μ g/mL (R2>0.999 9). RSDs of precision , reproducibility and stability tests were all lower than 2.00%. The recoveries were 96.23%-98.17%(RSD=0.96%-2.28%, n=6). Among 20 batches of samples ,the contents of above 4 kinds of phenolic acids were 0.385 3-1.891 9,0.018 0-0.129 5,2.569 5-10.676 0,0.563.5-1.860 5 mg/g. CONCLUSIONS : The established UPL C fingerprint could reflect the main chemical constituents of P. pedunculata . Phenolic acids could be used as the main evaluation indexes for the quality of P. petiolosa . The quality order of P. petiolosa from southwest China was Chongqing product>Sichuan product >Guizhou product.
6.Fingerprint Establishment of Rhizoma Ligustici from Different Origins and Their Chemometrics Analysis
Hui LIANG ; Xiaojun PAN ; Weisheng LYU ; Wenhui YANG ; Yuqin LUO ; Liye PAN ; Wenjie HUO ; Zhenyu LI
China Pharmacy 2021;32(19):2358-2362
OBJECTIVE:To establish UPLC fingerpri nt of Ligusticum sinense ,Ligusticum jeholense and Conioselinum vaginatium,and to conduct their chemometrics analysis so as to provide reference for the identification of Rhizoma Ligustici from different origins. METHODS :UPLC method combined with Similarity Evaluation System of TCM Chromatographic Fingerprints (2012 edition) were used to establish the fingerprints of Rhizoma Ligustici from different origins. Chromatographic peak identification and similarity evaluation were carried out. Cluster analysis (CA),principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA)were performed to analyze Rhizoma Ligustici from different origins,and screen differential components. RESULTS :Totally 13,11,11 characteristic peaks were identified in UPLC fingerprints of L. sinense ,L. jeholense and C. vaginatium ,respectively. Similarity evaluation showed that the similarity between C. vaginatium and L. jeholense were 0.312-0.541;that between C. vaginatium and L. sinense were 0.324-0.682;that between L. sinense and L. jeholense were 0.312-0.671,indicating there was great difference among Rhizoma Ligustici from different origins. CA,PCA and OPLS-DA showed that Rhizoma Ligustici from different origins were clustered into each category respectively ; chemical components represented by peak 10,peak 13,peak 12,peak 7 and peak 6 were differential components for Rhizoma Ligustici from 3 origins. CONCLUSIONS :The study establishes UPLC fingerprint of Rhizoma Ligustici from different origins , and screens 5 differential components ,which can be used to identify Rhizoma Ligustici from different origins.
7.Unqualified rate of anti-HIV detection in the laboratories from blood banks in Beijing-Tianjin-Hebei region
Wei ZHEN ; Hongwei GE ; Rui WANG ; Tong PAN ; Wei HAN ; Peng WANG ; Li YANG ; Shaoqiu SUN ; Xiao CAO ; Liye CUI ; Chao WEI ; Guijun YU ; Yunpeng XU ; Jinjuan FANG ; Caixia LIU ; Xuegang WANG ; Zhijun ZHEN ; Xiaojie LIU ; Wengong DU ; Lunan WANG ; Jiang LIU ; Hongjie WANG
Chinese Journal of Blood Transfusion 2021;34(4):371-376
【Objective】 To investigate the unqualified rate of anti-HIV detection of blood screening laboratories in Beijing-Tianjin-Hebei region, and explore the differences in anti-HIV detection ability and influencing factors in each laboratory. 【Methods】 Through filling questionnaires via e-mail, the anti-HIV ELISA unqualified rate and confirmed (WB) positive results (data) from January to December 2018 from 15 blood screening laboratories in Beijing-Tianjin-Hebei region were collected. Our laboratory was responsible for data collection and confirmation, and statistics software SPSS22.0 was used for analysis. 【Results】 1) There was a statistically significant difference among the unqualified rate of anti-HIV ELISA(6.77‱~35.71‱) and confirmed positive rate(0.60‱~3.56‱) in 15 blood screening laboratories in Beijing-Tianjin-Hebei region (P<0.05); 2) There were significant differencse among the ELISA unqualified rate and the confirmed positive rate of 8 reagents for anti-HIV detection(P<0.01), and the sensitivity of the 4th generation detection reagent and the imported reagent was higher than that of the 3rd generation reagent and the domestic reagent. The anti-HIV ELISA unqualified rate of R5 was the highest (19.08‱). 3)There were significant differences in the anti-HIV ELISA unqualified rate of R1, R2, R3, R5 and R7 reagents among different blood station laboratories(P<0.05), and there were no significant differences in the anti-HIV ELISA unqualified rate of R4, R6 and R8 reagents among different blood station laboratories(P>0.05). 4)The unqualified rate of anti-HIV ELISA of laboratories using different regents showed significant differences(P<0.05), except H, J, M. The unqualified rate of imported reagent was significantly higher than that of domestic reagents of laboratories using imported and domestic reagents combinations(P<0.05), except O. 62.5% (5/8) laboratories using domestic 3rd and 4th generation reagent combination showed significant differences in the unqualified rates among different reagents(P<0.05); 5) The positive rate of single-reagent(62.02%~95.45%)in 15 blood screening laboratories showed significant difference(P<0.001), and A was the lowest (62.02%). 【Conclusion】 The anti-HIV detection ability among 15 blood screening laboratories in Beijing-Tianjin-Hebei region is quite different. The application of different reagents is the main factor for the difference, and other factors such as personnel, instruments and test strategies also has a great impact on the detection of anti-HIV. It is still necessary to promote the process of homogenization of blood testing quality among blood screening laboratories in Beijing-Tianjin-Hebei region.
8.Comparison of Component Differences in Chebulae Fructus and Chebulae Fructus Immaturus Based on Chemical Pattern Recognition and Multi-index Quantitative Analysis
Caixian SUO ; Yunjing QIU ; Wenping WU ; Dongmei SUN ; Xiuzhi LI ; Liye PAN ; Guowei LI
China Pharmacy 2020;31(20):2512-2518
OBJECTIVE:To provide reference for the identification of Chebulae Fructus and Chebulae Fructus Immaturus . METHODS:UPLC method was adopted. The determination was performed on Waters Cortecs T 3 C18 column with mobile phase consisted of acetonitrile- 0.2% phosphoric acid solution (gradient elution )at the flow rate of 0.35 mL/min. The column temperature was 30 ℃,and the detection wavelength was set at 270 nm. The sample size was 1 μL. Using gallic acid as reference,UPLC fingerprints of 17 batches of Chebulae Fructus and 14 batches of Chebulae Fructus Immaturus were established and their similarity was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System (2012 edition). By comparing substance control , UV absorption spectrum and related literaturs ,common peaks were identified. PCA and PLS-DA were performed by using SPSS 20.0 and SIMCA 14.1 software. The contents of main difference components in Chebulae Fructus and Chebulae Fructus Immaturus were determined by above UPLC method and compared. RESULTS :There were 8 common peaks in UPLC fingerprint of Chebulae Fructus and Chebulae Fructus Immaturus ,i.e. chebulic acid (peak 1),gallic acid (peak 2),punicalagin A (peak 3),punicalagin B (peak 4),corilagin(peak 6),chebulagic acid (peak 7)and chebulinic acid (peak 8). The similarities of 17 batches of Chebulae Fructus were from 0.92 to 0.99,while 14 batches of Chebulae Fructus Immaturus were all above 0.99. The similarity of control fingerprint between Chebulae Fructus and Chebulae Fructus Immaturus was 0.909. PCA demonstrated the differences between Chebulae Fructus and Chebulae Fructus Immaturus . The results of PLS-DA were consistent with those of PCA ,and the variable importance in projection (VIP)values of peak 5,4,7,3 and 2 were above 1 in the PLS-DA model. In 31 batches of samples ,the contents of gallic acid (peak 2),punicalagin A(peak 3),punicalagin B (peak 4)and chebulagic acid (peak 7)were 2.63-10.31, 5.37-44.63,8.02-60.77,44.07-162.98 mg/g;RSDs were 40.14%, 47.91% ,53.97% ,36.22%(n=31). There was statistical significance in the differences of the mentioned 4 components between Chebulae Fructus and Chebulae Fructus Immaturus 719412818@qq.com (P<0.05). CONCLUSIONS :There are significant differences between Chebulae Fructus and Chebulae Fructus Immaturus gallic acid ,punicalagin A ,punicalagin B and chebulagic acid are the main difference components for identification.
9.Study on UPLC Fingerprint of Vaccariae Semen before and after Processing and Content Determination of Erythrine and Vaccarin
Siqiong CAO ; Wenping WU ; Yuqin LUO ; Ruirui MA ; Liye PAN ; Guowei LI ; Xiangdong CHEN
China Pharmacy 2020;31(19):2365-2370
OBJECTIVE:Compare the fingerprint difference of Vaccariae Semen before and after processed (stir-fried),and to determine the contents of erythrine and vaccarin before and after stir-fried. METHODS :UPLC method was adopted. The determination was performed on YMC Trait C 18 column with mobile phase consisted of acetonitrile-water (gradient elution )at the flow rate of 0.35 mL/min. The detection wavelength was set at 219 nm,and the column temperature was 35 ℃. The sample size was 1 μL. Using vaccarin as reference,the fingerprints of Vaccariae Semen crude product and its processed product (each of 17 batches,S1-S17,CS18-CS34) were drawn. The similarity evaluation and common peak identification were carried out by Similarity Evaluation System of TCM Chromatographic Fingerprint (2012 edition);cluster analysis ,principle component analysis (PCA)and factor analysis were performed by using SPSS 20.0 software. The contents of erythrine and vaccarin in Vaccariae Semen crude product and its processed product were determined by UPLC. RESULTS :There were 5 common peaks in UPLC fingerprints of 17 batches of Vaccariae Semen crude product and its processed product. The similarities were all higher than 0.99. Among them , 2 common peaks were identified ,i.e. erythrine ,vaccarin. Results of cluster analysis showed that S 1-S17 were clustered into one category and CS 18-CS34 were clustered into one category. Results of PCA and factor analysis showed that variance contribution rate of the first principle component was 76.418%;erythrine and vaccarin had higher loading on the first principal component (eigenvalues were 0.976 and 0.966,respectively). The linear ranges of above 2 components were 6.437-321.832 μg/mL and 7.729-386.437 μg/mL,respectively(r>0.999). The limits of detection and quantitation were 0.085,0.284 ng (crude product) and 0.739, 2.465 ng (processed product ), respectively. RSDs of precision ,reproducibility,stability(12 h)and durability tests were all lower than 3%(n=6 or n=5). E-mail:1083656123@qq.com Average recoveries were 96.42%(RSD=0.85%,n=6)and 99.13%(RSD=1.74%,n=6). The contents of the two components were 0.11%-0.20%,0.42%-0.63%(crude product )and 0.08%-0.11%,0.34%-0.50%(processed product ). CONCLUSIONS :UPLC fingerprint of Vaccariae Semen crude product and its processed product are established successfully. Although the chemical constituents in Vaccariae Semen are consistent before and after stir-fried ,the contents of erythrine and vaccarin are all decreased after stir-fried.
10.Dose reconstruction method for photon external radiation accident based on human voxel phantom
Yuan ZHAO ; Liye LIU ; Qinjian CAO ; Xueli HOU ; Hongjuan PAN ; Hua LI ; Yu WANG ; Yunshi XIAO ; Sanqiang XIA
Chinese Journal of Radiological Medicine and Protection 2019;39(8):624-628
Objective To study the method of dose reconstruction in human body under the photon external radiation accident condition,and to verify the accuracy of the method for the local dose distribution.Methods Based on the open source Monte Carlo tool kit Geant 4 and using the human voxel phantom recommended by ICRP Publication 103,the dose reconstruction method under the condition of external radiation accident was studied to evaluate the average absorbed dose,organ absorbed dose and local dose distribution.To validate the code,several irradiation experiments were implemented in some standard radiation fields by putting TLDs in the tissue equivalent physical phantom ART.A voxel phantom was used to reconstruct the radiation doses,which was created based on the CT scan image of the ART phantom with resolution of 1.57 mm× 1.57 mm× 10.00 mm.The result of experiment were compared with those of dose reconstruction simulation.Results The relative uncertainty of the measured values was 10.9%.The relative uncertainty of the dose reconstruction simulation values was 7.10% at the non-tissueinterface area and 16.6% at the tissue-interface area.For 451 measuring points,the average of the simulated value divided by the measured value was 0.972,with the standard deviation of 0.083 8.In the range of 0.95-1.05,0.90-1.10 and 0.80-1.20,and the proportions were 49.2%,79.4% and 96.4%,respectively.Conclusions The method of Monte Carlo dose reconstruction based on human voxel phantom meets the accuracy requirement of actual uses both at the whole body or organ level and at the local dose distribution level.It can be used as a powerful tool for dose assessment of the exposed people in an external radiation accidents and provide support for diagnosis and treatment.

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