Pharmacokinetic comparison between Tanreqing Capsules Substitute and Tanreqing Capsules in rats by LC-MS/MS / 中国中药杂志

Due to the limited resource of bear bile powder, the major raw material of Tanreqing Capsules(TRQ), cultured bear bile powder is used as a replacement to develop the Tanreqing Capsules Substitute(TRQS). An LC-MS/MS method was established in this study for simultaneous quantitation of 8 compounds from TRQS in rat plasma: tauroursodeoxycholic acid(TUDCA), taurocheno-deoxycholic acid(TCDCA), ursodeoxycholic acid(UDCA), chenodeoxycholic acid(CDCA), ferulic acid, wogonoside, baicalin, and forsythoside A. Thereby, the pharmacokinetic behaviors of TRQ and TRQS were evaluated. Concentration of endogenous compounds TUDCA, TCDCA, UDCA, and CDCA was determined with the stable isotope surrogate analytes: D4-TUDCA, D4-TCDCA, D4-UDCA, and D4-CDCA. Plasma samples were extracted by acetonitrile-induced protein precipitation. The LC conditions are as follows: Waters BEH C_(18) column(2.1 mm×100 mm, 1.7 μm), mobile phase of 10 mmol·L~(-1) ammonium formate aqueous solution(containing 0.01% formic acid) and acetonitrile-methanol mixture(1∶5). MS conditions are as below: multiple reaction monitoring(MRM), ESI~(+/-). Concentration of UDCA, CDCA, TUDCA, and TCDCA was corrected with a response factor, which is the ratio between the responses recorded for the surrogate and the authentic analyte at the equal concentration. Each of the plasma components showed good linearity(r > 0.995 1). Accuracy and precision met the criteria(inter-day RSD<7.0%, RE 89.98%-112.0%; intra-day RSD<12%, RE 90.41%-111.2%). The recovery was 64.83%-119.9% and matrix effect was 87.15%-113.8%. The validated method was applied for pharmacokinetic study of TRQS and TRQ(po, 0.94 g·kg~(-1)). There was no significant difference in C_(max) and AUC_(0-24 h) of baicalin, UDCA, TUDCA, and TCDCA between the two groups, indicating similar pharmacokinetic behaviors between TRQS and TRQ in rats.

Atomization Characteristics of Baicalin， Ursodeoxycholic Acid and Caffeic Acid in Tanreqing Inhalation Solution / 中国实验方剂学杂志

Objective:To investigate the effect of different angles of atomizer on the delivery rates and total delivery quantities of Tanreqing inhalation solution， so as to provide reference for the clinical use of this preparation. Method:Taking baicalin， ursodeoxycholic acid， caffeic acid as indexes， PARI Boy SX compression atomization inhaler （equipped with red core atomizing cup） and BRS2000 respiratory simulator were used， the effects of different angles of the atomizer （upper 15 degree， lower 15 degree， upper 30 degree， lower 30 degree， partial 15 degree， partial 30 degree， vertical） on the delivery rates and total delivery quantities of Tanreqing inhalation solution were investigated. The respiratory patterns of adults， children， infants and young children were selected to determine the delivery rates and total delivery quantities of three components in Tanreqing inhalation solution. Result:In the same atomization time， the delivery rates and the total delivery quantities of caffeic acid and ursodeoxycholic acid in Tanreqing inhalation solution were not significantly affected by the atomizer from different angles， but significantly affected baicalin. At the vertical angle， the delivery rate and total delivery quantity of baicalin were higher than the other angles. Under different respiratory modes， there were significant differences in the delivery rates and total delivery quantities of these three components in the inhalation solution. Compared with other respiratory modes， the delivery rates and total delivery quantities of baicalin， ursodeoxycholic acid and caffeic acid were the highest in the adult respiratory mode， with delivery rates of （555.5±16.61）， （226.3±6.54）， （26.1±0.32） μg·min-1 and total delivery quantities of （4 001.1±82.97）， （1 754.9±63.73）， （167.6±1.42） μg， respectively. Conclusion:The use angle of atomizer has a certain effect on the delivery rate and total delivery quantity of Tanreqing inhalation solution， so it is suggested that the vertical angle should be kept as far as possible in clinical use. Under the four respiratory patterns， the delivery rate and total delivery quantity of Tanreqing inhalation solution are different， suggesting that the atomization dose or atomization time should be adjusted according to the respiratory characteristics of the patients to ensure the safety and effectiveness of clinical medication.

Systematical analysis of multiple components in drainage bear bile powder from different sources / 中国中药杂志

Bear Bile Powder contains bile acids, protein, amino acids, bilirubin and microelements and other compounds. Among them, the bile acids are the most active components. Currently, there are many studies on bile acids, but few reports on other components. Therefore, the purpose of this study is to carry out a systematical analysis of multiple components in drainage Bear Bile Powder from different sources. Bilirubin and protein were quantified by microplate spectrophotometer. The contents of bile acids and amino acids were determined by liquid chromatography coupled with mass spectrometry (LC-MS). The contents of microelements were determined by inductively coupled plasma mass spectrometry (ICP-MS) The result indicated that among 20 batches of bear bile powder from different sources there is high similarity (0.922-0.977). Tauroursodeoxycholic acid (TUDCA) and taurochenodeoxycholic acid (TCDCA) were the two most abundant components. The total contents of them were 41%-59% and met the current standard for quality control of bear bile powder. However, significant differences were found in their contents among samples from different sources. Besides, bilirubin, protein, amino acids and microelements also contributed to the differentiation of samples from different sources. The main components of bear bile powder from the different sources were with satisfactory similarity. But bile acids, bilirubin, protein, amino acids and microelements all contributed to the different among samples. Our present study provided a systematical approach for the better quality control and evaluation of bear bile powder.

Identification of bear bile powder and its adulterants by using DNA barcoding technology / 中国中药杂志

To identify the precious bile powder and its adulterants by DNA barcoding, and establish its standard experimental process to ensure the safe and effective utilization. Total twelve sequences from samples of bear bile powder which come from Ursus thibetanus for DNA extraction, PCR(polymerase chain reaction) and sequence, then using CodonCode Aligner V 7.0.1 shear primer region to obtain COI sequence. The COI sequences of U. arctos and their adulterants were obtained from GenBank. MEGA7.0 software was applied for analyzing mutation, calculating intraspecific and interspecific K2P(Kimura 2-Parameter) genetic distance and constructing the Neighbor-joining tree(NJ). The results showed that the maximum K2P genetic distance of bear bile powder of U. thibetanus and U. arctos are far less than minimum K2P genetic distance within its adulterants species, and the results of NJ tree demonstrated that each species could be distinguished from the counterfeits obviously. DNA barcoding is a safe, convenient and reliable technique for species identification, and it is important to establish the standard sequence of COI sequences for animal medicines.

Primary exploration of key influential factors recognization method of Tanreqing injection / 中国中药杂志

To recognize the key influential factors during the liquid preparation process of Tanreqing injection, the near infrared(NIR) spectra of the raw materials and the operating parameters of 24 batches of physical manufacturing were recorded as independent variables, and the total soluble solids contents and the light inspection acceptance rate of the final products were collected as dependent variables. The calibration models were developed using the partial least-square regression (PLSR) method, and the correlation coefficients between the independent variables and the dependent variables were calculated. For the quantitative models, the correlation coefficients for the calibration and inner cross validation of total soluble solids contents and the light inspection acceptance rate reached 0.911 9, 0.724 2 and 0.873 8, 0.795 9, respectively. Using the correlation coefficients diagrams, several key influence factors were preliminarily determined, and the physical significance were analyzed combined with production experience. This work demonstrated that NIR spectroscopy with PLSR algorithm could be used for the key influential factors recognization during the liquid preparation process of Tanreqing injection and can be popularized to solve similar problems..

Screening and identification of multiple components in Tanreqing injection using RP-HPLC combined with DAD and ESI-TOF/MS / 中国天然药物

A liquid chromatography coupled with diode array detector (DAD) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF/MS) method was developed for the screening and identification of the multiple components in Tanreqing injection, a well-known Chinese medicine injection in China. By combining the DAD spectrum and the accurate mass measurement of ESI-TOF/MS, twelve components in Tanreqing injection were identified. This study contributes to clarifying the nature of Tanreqing injection, and provides an effective and reliable process for the comprehensive and systematic characterization of complex traditional Chinese medicine preparations.

NIRS-based total quality control system construction of Tanreqing injection and relative thinking / 中国中药杂志

In this paper, NIRS (near infrared spectroscopy)-based total quality control system for the Tanreqing injection was introduced briefly. By analyzing and summing up the significance and difficulties, several important problems of the practical applications which need urgent solutions are proposed. And also the applicationprospect of NIRS is fully discussed and put forward in the end.

Demonstration of nested case-control study design in mechanisms research of allergic reaction of tanreqing injection / 中国中药杂志

The paper is focused on the clinical applications of Tanreqing injection after listing, detecting and analyzing the related blood indicators of patients with allergic reactions based on prospective, multi-center, large sample, registration-type clinical safety monitoring nested case-control study (NCCS) to explore the possible mechanisms of allergic reaction of Tanreqing injection, 3 006 patients cases used with Tanreqing injection were monitored, including 3 cases of adverse reactions and 2 cases of allergic reactions. Each patient of allergic reactions, according to the ratio of 1:4 matches four cases of not adverse reactions as a control group of patients, while 5 healthy and 5 cases of volunteers into the healthy group. We examined the correlation detection of cases of allergic reactions among groups such as T-IgE, IgA, IgG, C3, C4, IFN-g, IL-2, IL-4, IL-6, IL-10. Allergic reactions of Tanreqing injection may be mediated by IgE as type I based on existing research data. This results and conclusions will promote the justifiability and safety of clinical applications.

Technological parameter optimization method for washing of coarse bear gall powder extracts based on NIR spectroscopy / 中国中药杂志

During the washing process of coarse bear gall powder extracts, it is necessary to adjust the amount of ethyl acetate according to the properties of raw materials, which aims to improving the yield and purity of the final product. In the research, using NIR spectra to reflect the comprehensive properties of coarse bear gall powder extracts, the process is optimized in a flexible way. Forty batches experiments are designed according to the weight ratio of ethyl acetate and coarse extracts of bear gall powder. The NIR spectra of the coarse extracts of bear gall powder are collected and processed using principal component analysis (PCA) method. The first 8 principal components combined with the amount of the ethyl acetate are used as the input variables, and calibration models are established to predict the yield and purity of the final product 30 batches are used as calibration set, which is used to establish the models, and other 10 batches are used as validation set, which is used for the performance appraisal of the established models. The correlation coefficients of the calibration, inner cross-validation and external validation for the purity model are 0.902, 0.896 and 0.883, respectively, and the RMSEC, RMSECV and RMSEP are 1.22%, 1.48% and 1.59%, respectively. The correlation coefficients of the calibration, inner cross-validation and external validation for the yield model are 0.921, 0.859 and 0.916, respectively, and the RMSEC, RMSECV and RMSEP are 1.39%, 1.65% and 1.53% respectively. This work demonstrated that NIR spectra combined with technology parameter could be used to predict the yield and purity of the final product. Using the established models, the most appropriate amount of the ethyl acetate can be determined according to the properties of the coarse bear gall powder extracts, and the yield and purity of the final product can be improved.

Quality control of Lonicerae japonicae flos with standard reference extract / 中国中药杂志

The high price of the reference substances is an obstacle for the HPLC analysis of Lonicerae Japonicae Flos. To solve this problem, a new method based on the standard reference extract (SRE) was proposed. In this study, the extract of Lonicerae Japonicae Flos was calibrated, and the long-term stability was investigated. Different concentration solutions of SRE were prepared for establishment of the calibration profiles, and 6 organic acids were determined. T-test was used for the comparison of the determination results via reference substances and SRE, and the results demonstrated that there is no significant difference between the two methods. The presented method can be used for the quality control of Lonicerae Japonicae Flos, and will also offer reference to resolve similar problems.

Effect of Tanreqing Injection in various formula compatibilities on pharmacokinetics of main components / 中草药

Objective: To study the pharmacokinetic effect of three active markers, baicalin (BC), ursodeoxycholic acid (UDCA), and chenodeoxycholic acid (CDCA) in plasma of rats after iv administration of Tanreqing Injection (TI) in various compatibilities, which contained scutellaria extraction (SE), bear gall powder extraction (BE), Cornu caprae hircus extraction (CE) and Lonicerae Flos extraction (FE), including SE, BE, SE-BE (SE and BE), SE-CE (SE and CE), SE-BE-CE (SE, BE and CE), and SE-BE-CE-FE (SE, BE, CE, and FE):. To discuss the rationality of TI compatibility, the effects of TI in various compatibilities on the pharmacokinetics of BC, UDCA, and CDCA in plasma of rats were investigated. Methods: Thirty-six experimental rats were randomly divided into six groups, and treated with SE, BE, SE-BE, SE-CE, SE-BE-CE, and SE-BE-CE-FE. After the simultaneous extraction of the three major bioactive components in plasma of rats, the concentration of BC was determined using LC-UV method, and UDCA as well as CDCA was determined using LC-MS method. The experimental data were analyzed by WinNonlin 6.3 software and the pharmacokinetic parameters of BC, UDCA, and CDCA in these recipes were evaluated. Results: The pharmacokinetic parameters of UDCA and CDCA did not change after iv administration of TI in various compatibilities. However, the TI in various compatibilities increased the AUC of BC after iv administration. The change degrees were SE < SE-CE < SE-BE-CE < SE-BE-CE-FE but with no statistical significance. The pharmacokinetic parameters of UDCA and CDCA did not obviously change, which indicated that the different compatibilities did not change the pharmacokinetic behavior of UDCA and CDCA. Conclusion: TI in various compatibelities could increase the exposure of BC following the iv administration, but could not affect the pharmacokinetic behaviors of UDCA and CDCA.