Optimization of TLC identification for Cirsii Herba and research on changes in quality properties "carbonizing retains characteristics" of its charred product / 中成药

AIM To optimize the TLC identification method for Cirsii Herba and to study the changes in quality properties"carbonizing retains characteristics"of Cirsii Herba Carbonisata.METHODS After the improvement of thin layer plate and solvent in the TLC identification based on the Chinese Pharmacopoeia 2020 Edition for Cirsii Herba,Cirsii Herba Carbonisata had its TLC identification method and UPLC fingerprint established for the determination of its content of buddleoside and acacetin as well.RESULTS We used silica gel G plate,and the solvent of toluene-acetone-formic acid-methanol(6 ∶ 3 ∶ 0.5 ∶ 2.5)for TLC identification of Cirsii Herba.The content variations of buddleoside and acacetin in Cirsii Herba and its differently charred products were consistent with the result of the TLC identification.CONCLUSION The improved TLC identification method for Cirsii Herba is of lower cost and less solvent toxicity compared to the method in the Chinese Pharmacopoeia 2020 Edition,and can identify the changes in quality properties"carbonizing retains characteristics"of differently charred Cirsii Herba,therefore it can be used to control the quality of Cirsii Herba Carbonisata in the market.

Chemical Composition Analysis of Platycladi Cacumen Before and After Being Carbonized Based on Identification by UHPLC-QTOF-MS/MS / 中国实验方剂学杂志

Objective:To identify the chemical constituents of Platycladi Cacumen<italic> </italic>before and after being carbonized. Method:Chemical constituents in 3 batches of Platycladi Cacumen and its carbonized products<italic> </italic>were identified and compared by ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry （UHPLC-QTOF-MS/MS）. Chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.7 μm） with 0.1% formic acid aqueous solution （A）- acetonitrile （B） as mobile phase for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B； 18-22 min， 100%B）. The flow rate was 0.4 mL·min^-1 and the injection volume was 5 μL. Mass spectrometry was performed by an electrospray ionization， and the primary and secondary mass spectrometry data were collected with the full scan mode of positive and negative ions， the peaks containing MS/MS data were identified by self-established secondary mass spectrometry database and corresponding fragmentation law matching method. Result:A total of 77 and 76 substances with the same change trend were identified under positive and negative ion modes. After being<italic> </italic>carbonized， the disappeared components of Platycladi Cacumen were mainly amino acids， ketone aldehydes and other volatile components. Among newly produced components， there were 6 kinds of flavonoid aglycones （rhamnetin， 6，7，3'-trihydroxyflavone， 3，6，3'-trihydroxyflavone， 4'-hydroxy-2'-methyl-3，4，5-trimethoxychalcone， herbacetin and 3'，5'-dimethoxy-3，5，7，4'-tetrahydroxyflavone）， 3 kinds of coumarins （7-hydroxycoumarin， 7，8-dihydroxycoumarin and 8-acetyl-7-hydroxycoum-arin） and 3 kinds of benzoic acids （3-methylcatechol， pyrocatechol and chromone-3-carboxylic acid）. There were a total of 40 flavonoids （quercitrin， quercetin， kaempferol， etc.） among these identified chemical constituents. Conclusion:There are significant quantitative and qualitative changes in the chemical compositions of Platycladi Cacumen after being carbonized. The flavonoids， the identified main active ingredients， can provide data reference for further study on the material basis of efficacy changes of Platycladi Cacumen<italic> </italic>before and after being carbonized.

Analysis of volatile organic compounds (VOCs) fingerprint of raw and honey-fried licorice based on headspace-gas-chromatography ion-mobility spectrometry (HS-GC-IMS) / 中国中药杂志

Licorice is one of the most commonly used traditional Chinese medicine. In clinic, raw licorice and honey-fried licorice are used in medicines, with the main effects in clearing away heat and detoxifying, moistening lungs and removing phlegm. Honey-fried licorice has effects in nourishing the spleen and stomach and replenishing Qi and pulse. Because traditional Chinese medicine exerts the effects through multiple components and multiple targets, the index components used in the quality evaluation of licorice are often difficult to reflect their real quality. In addition, most of studies for the quality standards have shown that honey-fried licorice are the same as licorice, with a lack of quality evaluation standards that can demonstrate their processing characteristics. The quality of medicine is directly related to its clinical efficacy, so it is necessary to establish a more effective quality control method. Licorice has a beany smell, which is one of the main quality identification characteristics. In this study, by taking advantage of the odor characteristics, a headspace-gas chromatography-ion migration mass spectrometry technology was used to establish a quality evaluation method. A total of 76 volatile components were identified. Through the dynamic principal component analysis, 7 kinds of volatile substances in raw licorice and 13 kinds of volatile substances in honey-fried licorice were statistically obtained, and could be taken as index components for the quality evaluation of raw and honey-fried licorice, respectively. This study could help realize the combination and unification of modern detection and traditional quality evaluation methods, and make a more realistic evaluation for the quality of licorice.

Primary study on scientific connotation "carbonizing retains characteristics" of Platycladi Cacumen / 中草药

Objective: To study the scientific connotation "carbonizing retains characteristics" of Platycladi Cacumen (PC). Methods: Chemical constituents of PC and Platycladi Cacumen Carbonisata (PCC) of different carbonizing degree were compared by HPLC characteristic chromatogram, and the chromatographic peaks were assigned; PC and PCC of different carbonizing degree and the characteristic components changed before and after being carbonized were used to compare the impact to hemostasis in vitro and inhibition of zebrafish cerebral hemorrhage. Results: Chemical compositions of PC were affected by different carbonizing degree, when carbonizing degree is moderate, the content of myricitrin, quercitroside, isoquercitroside, amentoflavone and hinokiflavone original in PC were significantly reduced and the amount of quercetin and kaempferol which were newly produced were higher. In vitro hemostatic experiments showed that compared with the blank group, the APTT and FIB of PC which was carbonized moderately were significantly different (P < 0.01) and TT was significantly different (P < 0.05). It also had obvious inhibition effect on zebrafish cerebral hemorrhage when the concentration of PC carbonized moderately was 50 μg/mL (P < 0.01). The enhanced hemostasis was significantly related to newly produced compositions quercetin and kaempferol. Conclusion: Chemical compositions of PC changed significantly after being carbonized and the hemostatic effect was enhanced,which were related to the processing degree. The scientific connotation and traditional processing requirements of "carbonizing retains characteristics" was preliminarily analyzed when the carbon medicine was processed by this study, which provides a certain idea for the research on traditional processing theory of carbon medicine.

Study on internal and external quality control methods of Platycladi Cacumen Carbonisata based on QAMS and color recognition / 中国中药杂志

In order to effectively solve the over-processing problem of Platycladi Cacumen Carbonisata, which was commonly used as a hemostatic drug in clinical application, we used the quantitative analysis method of multi-components by single marker(QAMS) in this study, with quercetin as internal reference to simultaneously determine the content of six flavonoids which can be used to control the internal quality of Platycladi Cacumen Carbonisata. Based on the comparison of QAMS and external standard method(ESM) results, the limit standards of contents were established as follows: isoquercitroside ≥0.002 0%, quercitroside ≥0.050%, quercetin ≥0.030%, kaempferol and amentoflavone both ≥0.010%, hinokiflavone ≥0.050%. Based on the color detection of Platycladi Cacumen and Platycladi Cacumen Carbonisata with different processing degrees, the law of influence of different processing degrees on the color of Platycladi Cacumen Carbonisata was found. A new external quality standard of Platycladi Cacumen Carbonisata was established by fitting curve of color recognition for the external quality control, based on which the standard ranges of ΔL~*, Δb~* and ΔE were-50.00--44.00, 6.00-11.00 and 45.00-50.00 respectively. Effective combination of established internal and external quality control standards by this study can be used to evaluate the processing degree and quality of Platycladi Cacumen Carbonisata more comprehensively and objectively, which can guarantee its clinical efficacy. At the same time, this study also provides reference and basis for further improving the quality control standard of Platycladi Cacumen Carbonisata.

Analysis of Quality Control Standard of “Carbonizing Retains Characteristics” Based on Quality Investigation of Commercial Platycladi Cacumen Carbonisata Pieces / 中国实验方剂学杂志

Objective: To investigate the quality of Platycladi Cacumen carbonisata pieces in the market,then to explore control standard of "carbonizing retains characteristics" of Platycladi Cacumen.Method: The properties,water content and alcohol extract content of commercial Platycladi Cacumen carbonisata pieces were determined by the method in the 2015 edition of Chinese Pharmacopoeia.Thin layer chromatography(TLC) identification of quercetin and contents of quercetin and quercitin were determined.Result: The carbonizing degree of 9 batches of Platycladi Cacumen carbonisata pieces were heavier than which is specified in the 2015 edition of Chinese Pharmacopoeia.The conditions of TLC identification in the 2015 edition of Chinese Pharmacopoeia should be revised to avoid stratification;the carbonizing degree could affect the contents of quercetin and quercitin in Platycladi Cacumen carbonisata pieces;the simultaneous detection of quercetin and quercitin can be used as one of the quality control indexes of this carbonisata pieces.Conclusion: Most of commercial Platycladi Cacumen carbonisata pieces have the quality problem of excessive carbonizing degree,the established quality control standard of "carbonizing retains characteristics" can be used to effectively ensure the quality of Platycladi Cacumen carbonisata pieces.

Investigation on quality of Haematitum and Fluoritum pieces in venalicium and study on related problems of mineral drugs / 中国中药杂志

The main problems and solutions of mineral drugs in clinical use were studied based on the investigation on the quality of Haematitum and Fluoritum pieces in venalicium. The outward appearance and intrinsic quality of Haematitum and Fluoritum pieces in venalicium were studied by the requires which were in the first part of Chinese Pharmacopoeia published in 2010. The outward appearance and intrinsic quality of mineral drugs had large differences, the disqualification rate was 41.67% in 12 batches of Haematitum pieces and the disqualification rate was 53.85% in 13 batches of Fluoritum pieces. The crushing granularity of mineral drugs should be defined, the quality standards should be further improved. The drug adiministration ought to strengthen inspection and supervision in order to ensure the stability and reliability of the clinical efficacy.