OBJECTIVE To develop the limited standard and supporting detection methods of commonly used pesticides in Lycium barbarum L. (L. barbarum). METHODS In previous research, the “the Principle of Conversion of Limit Standards of Traditional Chinese Medicine in <GB 2763 national food safety standard-maximum residue limits for pesticides in food>” have been drafted. In this research, the L. barbarum was used as the research object to study the conversion scope, risk assessment, detection methods and conversion judgment principle. RESULTS A method for the determination of 13 pesticide residues in the L. barbarum was established by using acetonitrile extraction and HLB column purification. And the GC-MS/MS and LC-MS/MS technology were used to detect the pesticide residues. In addition, the maximum residue limit of 13 pesticides in L. barbarum was finally formulated by sample determination. CONCLUSION The specific implementation steps are provided for the conversion of GB 2763 standard to traditional Chinese medicine standard in this study, and it is of great significance for promoting the connection between planting and circulation supervision of L. barbarum.

OBJECTIVE To screen the commonly used pesticides in Dendrobium officinale to understand their pesticide residues, and explore the establishment of a method for determining the transformation of pesticide residues in Dendrobium officinale and the development of limit standards in accordance with the requirements of GB 2763 Food Safety Standards for Maximum Residue Limits of Pesticides in Food and the Chinese Pharmacopoeia. METHODS LC-MS/MS methods were used to screen and risk assess 6 transformation pesticides in 53 batches of Dendrobium officinale, and the pesticide registration status and the maximum residue limit standards for Dendrobium officinale (dry) in GB 2763 were used to determine the indicators for the transformation of pesticides. RESULTS The pesticides in 53 batches of Dendrobium officinale were analyzed. The transformation pesticide were confirmed in accordance with GB 2763 and the requirements of the Chinese Pharmacopoeia 2020 Edition, and a method for determining the transformation of pesticide residues in Dendrobium officinale was established, along with the development of limit standards. CONCLUSION This study screened 40 commonly used pesticides and 6 transformation in 53 batches of Dendrobium officinale, established a method for determining the transformation of pesticide residues in accordance with GB 2763, and developed limit standards. This study is helpful for understanding the risk of pesticide residues in Dendrobium officinale, regulating the use of pesticides during its cultivation process, and improving its quality standards.

OBJECTIVE To establish a method for the transformation of some commonly used pesticide limit standards in Chrysanthemi Flos based on ‘GB 2763 National Food Safety Standard-Maximum Residue Limits for Pesticides in Food’. METHODS The indexes of Chrysanthemi Flos (dry) in GB 2763 were sorted out, and the quasi-transformation pesticides were determined as pymetrozine, imidacloprid, boscalid and fluazinam according to the transformation principle. The UPLC-MS/MS method was used to establish the transformation method of Chrysanthemi Flos, and the methodological verification was carried out. RESULTS The four pesticides had good linear relationship in the range of 1-50 ng·mL^-1, the correlation coefficients were 0.999 9, the average recovery was 77.9%-95.5%, and the RSD was 1.7%-6.8%. The overall detection rate of the samples was 60%, and the pesticide with the highest detection rate was imidacloprid. The maximum detectable amount was 1 mg·kg^-1 (imidacloprid), and the results did not exceed the maximum residue limit specified in GB 2763. The risk assessment system of pesticide residues in traditional Chinese medicine was used to evaluate the chronic risk of the four pesticide limit values to be transformed. The results showed that the chronic risk of fluazinam was greater than 1, and the chronic risk quotients of the other three pesticides were less than 1. Therefore, fluazinam is not converted temporarily, and only pymetrozine, imidacloprid and boscalid are converted. CONCLUSION The three limit standards of pymetrozine, imidacloprid and boscalid in Chrysanthemi Flos were transformed. The established limit standard transformation method is simple and accurate, which is of great significance for standardizing the use of pesticides and medicinal safety in the process of chrysanthemum planting.

OBJECTIVE To establish a method for simultaneous determination of registered pesticides, prohibited pesticides, and proposed new prohibited pesticide residues in Fritillaria Medicinal Herbs, and propose reference limits. METHODS The detection method for 95 pesticide residues in Fritillaria Medicinal Herbs was established using GC-MS/MS and LC-MS/MS. The prepared samples were extracted with acetonitrile and then purified with the QuEChERs method. A total of 70 batches of samples were detected by the established method. RESULTS According to the GB 2763 conversion guidelines, imidacloprid and avermectin in Fritillaria Medicinal Herbs should be converted to the maximum residue limit (MRL) standards. Additionally, reference limit values are proposed for the 18 prohibited pesticides that were previously suggested for addition. CONCLUSION The study of Fritillaria Medicinal Herbs aims to offer guidance and inspiration for the transformation of traditional Chinese medicine standards and the expansion of the list of banned pesticides. It also provides a reference for the regulatory policies and technological advancements related to pesticide residue management in Fritillaria Medicinal Herbs and other traditional Chinese medicines.

OBJECTIVE To carry out screening of pesticide residues in Panax notoginseng for the pesticide residues commonly used in Panax notoginseng,understand the pesticide residues situation and carry out the related risk assenssment study. METHODS GC-MS/MS and LC-MS/MS methods were used to establish detection methods for commonly used pesticides in Panax notoginseng, as well as those regulated by laws and regulations; the methods were used to conduct a comprehensive screening and risk assessment of the collected Panax notoginseng samples; and the indexes of the transformed pesticides were confirmed in accordance with the pesticide residue limits of Panax notoginseng in the GB 2763 standard. RESULTS In this study, the residue determination methods for about 40 commonly used pesticides and pesticides regulated by regulations in Panax notoginseng were successfully established, and the corresponding limits were set according to the GB 2763 standard. Eight pesticides including tebuconazole, phenyl ether metronidazole and carbendazim were finally recognized as transformed pesticide indicators. CONCLUSION The pesticide residue detection method for Panax notoginseng established in this study is characterized by simple and rapid operation, high specificity and high sensitivity, and the related risk assessment and limit setting provide technical support for the improvement of the quality standard of Panax notoginseng.

In recent years, in the process of drug research and development, insoluble drugs are faced with the problem of low bioavailability and poor absorption effect. Drug nanocrystals have attracted much attention due to their nano size and unique physical and chemical properties, which can effectively increase drug solubility and permeability, and have advantages such as low dosage of excipients, high drug load and few adverse reactions. However, instability limits their further development. Multieffect stabilizers can significantly improve their stability through steric hindrance, electrostatic repulsion and solvation. The author reviews the main characteristics and limitations of drug nanocrystals, the classification of multi-effect stabilizers and their applications in vivo and in vitro by reviewing and sorting out relevant literature, in order to provide reference for the research and development of drug nanocrystals.

OBJECTIVE To develop and validate a peptide mapping method of an anti-CD33 monoclonal antibody. METHODS Different types of chromatographs (HPLC, UPLC) and different mobile phase systems (formic acid, trifluoroacetic acid) were used for peptide map detection of anti-CD33 antibody. The signature peptide segments were localized using synthetic CDR peptide of the antibody and the localization results were confirmed by mass spectrometry. Based on the relative retention time (RRT), the specificity, precision, and robustness of the method were validated according to the Pharmacopoeia of the People's Republic of China (ChP, 2020). RESULTS The separation time of the peptides by UPLC was shorter than that by HPLC, and the degrees of separation with trifluoroacetic acid in the mobile phase were higher than that with formic acid. The identification results of the signature peptide segment using the maps of synthetic peptide segments were consistent with the results of mass spectrometry. The specificity validation demonstrated that the formulation blank and sample solution blank did not interfere with the detection of signature peptide segments, and there were significant differences between peptide mapping results of different antibodies. The repeatability validation showed that the RSDs (RRT) of signature peptide segments between six parallel samples were 0.01%-0.05%; the intermediate precision validation proved that the RSDs (RRT) of signature peptide segments for different analysts were 0.04%-0.32%; the robustness validation exhibited that the RSDs (RRT) of signature peptide segments were 0.02%-0.09% under different enzyme treatment conditions and 0.36%-1.43% under different chromatographic conditions. Within 25 h in detection, the RSDs (RRT) of the signature peptide segments were 0.01%-0.04%. CONCLUSION This study uses synthetic peptide segments for peptide localization in peptide mapping detection and uses relative retention time to determine the results, which provide a new approach for biopharmaceutical peptide mapping detection.

OBJECTIVE To investigate the feasibility of performing identification of Citri Sarcodactylis Fructus (CSF) and Citri Fructus (CF) based on high performance liquid chromatography (HPLC) fingerprint coupled with multivariate data analysis. METHODS HPLC method was applied to establish the fingerprints of CSF and CF. Similarity evaluation of 12 batches of CSF samples and 10 batches of CF samples was performed by Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2004A edition). Hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used for data analysis. RESULTS HPLC fingerprints of CSF and CF were established, respectively. The fingerprint similarities of CSF samples, as well as the fingerprint similarities of CF samples, were all greater than 0.9. The similarities between the fingerprints of CSF samples and the control fingerprint of CF, as well as the similarities between the fingerprints of CF samples and the control fingerprint of CSF, were all less than 0.9, which indicated significant differences in chemical composition between CSF and CF. The total 22 samples were divided into two groups by HCA, PCA and OPLS-DA, which was consistent with the two varieties. Three differential markers were screened by OPLS-DA and two of them were identified as 5, 7-dimethoxycoumarin and hesperidin by reference substances. The statistical analysis indicated that the ratio of absolute peak area (APA) of hesperidin and 5, 7-dimethoxycoumarin can distinguish CSF and CF from each other simply and accurately. CONCLUSION HPLC fingerprint coupled with multivariate data analysis can be used to identify CSF and CF.

OBJECTIVE To investigate the ameliorative effect of Guifu Dihuang Wan (GFDHW) on intestinal lipid absorption dysfunction in aging mice and its potential mechanisms. METHODS Twenty-six SPF male C57BL/6J mice aged 11 months were raised to 21 months and divided into an aging model group (Model group) and a GFDHW group (1.17 g·kg^-1). The GFDHW group received medication via feed administration for 3 months, with free access to water during the administration period, while the Model group was fed normal feed and had free access to water. Weekly changes in body weight were recorded for each group of mice, and specimens were collected at 24 months of age. Nine SPF male C57BL/6J mice aged 8 months were used as the youth control group (Con group) and were acclimatized for 1 week before specimen collection. Prior to specimen collection, all groups of mice were fasted for 24 hours, and 0.2 mL of olive oil was administered by gavage to each group of mice at the end of the fasting period, followed by gavage of 0.2 mL of semi-solid nutrient solution 30 minutes before specimen collection. Small intestine propulsion tests were conducted to assess the small intestine motility of each group of mice. Biochemical reagent kits were used to measure the serum levels of triglycerides (TG), total cholesterol (TC), and low-density lipoprotein (LDL) in each group of mice. Transmission electron microscopy was used to observe changes in the number of lipid droplets in intestinal epithelial cells of each group. The calcium cobalt method was used for alkaline phosphatase staining to detect intestinal alkaline phosphatase (IAP) activity in each group of intestines. Hematoxylin-eosin (HE) staining was used to observe the morphology of intestinal epithelia in each group. Immunohistochemical staining was performed to detect the expression of Olfm4, Lgr5, PCNA, Notch1, Hes1 and Villin in the small intestines of each group of mice. Western blot analysis was conducted to determine the expression levels of lipid absorption-related molecular proteins in the small intestines of each group of mice. RESULTS Compared with the Con group mice, the Model group mice exhibited faster weight loss, decreased small intestine motility, increased serum TG, TC, and LDL levels, and increased number of lipid droplets in intestinal epithelial cells. The GFDHW group mice showed improved small intestine motility and decreased serum TG, TC, and LDL levels. HE staining revealed disordered arrangement and fracture of small intestinal villi, shortened villus length, decreased crypt depth, and reduced crypt numbers in Model group mice, whereas the GFDHW group mice exhibited orderly and continuous arrangement of small intestinal villi, increased villus length, higher crypt depth, and increased crypt numbers. The immunohistochemical results showed that compared to the Con group mice, the expression of Olfm4, PCNA, Notch1, Hes1, and Villin proteins in the small intestine of the Model group mice decreased, while the levels of these proteins increased in the GFDHW treated group mice. Western blot analysis revealed that compared to the Con group mice, the expression of CD36, FATP4, SR-BI, ACAT2, FABP1 and MTTP proteins in the small intestine of the Model group mice decreased, whereas the levels of these proteins increased in the GFDHW treated group mice. CONCLUSION GFDHW improves lipid absorption function in aging mice by upregulating Notch1 expression, promoting differentiation of intestinal stem cells into absorptive cells, and increasing the expression levels of related molecular proteins in the lipid absorption pathway.

OBJECTIVE To prepare risedronate sodium (RIS)-loaded dissolving microneedle (DMN) and evaluate its efficacy in preventing postmenopausal osteoporosis. METHODS The preparation process of RIS-DMN was optimized by Box-Behnken design of response surface methodology. The appearance, solubility, mechanical property, safety and transdermal effect of RIS-DMN were characterized by scanning electron microscope, intradermal dissolution test, puncture test, skin barrier recovery test and in vitro permeation test. The pharmacodynamic evaluation of RIS-DMN was performed in ovariectomized osteoporosis model rats. RESULTS The optimal formulation were determined to be 45% for solute (mixed with 1:0.86 PVP K30 and CS) and 55% for solvent. It was found that the RIS-DMN have good physical characteristics and properties, and showed great effects in regulating the level of Ca²⁺, P³⁺and alkaline phosphatase (ALP). Meanwhile, the RIS-DMN showed great effects in repairing bone microstructure and improving bone density in ovariectomized osteoporosis model rats, as similar as oral administration. CONCLUSION The RIS-DMN has stable quality, convenient use and precise efficacy, shows great potential in the treatment of postmenopausal osteoporosis.

OBJECTIVE To establish an analysis method of the charge heterogeneity of human urinary kininogenase (HUK) by using image capillary isoelectric focusing (iCIEF) and complete the methodological verification. METHODS The 35 μL 1% methyl cellulose, 10 μL 200 mmol·L^-1 IDA, 4 μL pharmalyte electrolyte, 48 mg urea, 0.5 μL pI marker with pI 6.15 and 7.05 were added in the sample solution. The focusing condition was pre-focusing voltage 1 500V, duration 1 min, focusing voltage 3 000 V, duration 6 min. RESULTS The optimized method has stable baseline, and the target protein was significantly different from the unrelated protein, The recovery rate of accuracy verification was within 90%-110%, and the linearity verification result had r² of 0.995 7, The RSD of each isomer pI in the repeatability verification was less than 0.2%. The limit of quantification was 0.013 mg·mL^-1. The concentration of urea durability, IDA durability and the electrolyte pharmalyte durability are good. Using this method, the charge isomers of HUK from different manufactures were analyzed effectively. CONCLUSION The developed iCIEF method has good specificity, precision, linearity and durability, and can solve the problem of charge heterogeneity evaluation of protein products, which is of great significance to the quality control of such products from the perspective of charge heterogeneity.

OBJECTIVE To establish a quantitative nuclear magnetic resonance coupled with high performance liquid chromatography (qNMR-HPLC) technique for the rapid determination of cilostazol impurity Ⅰ correction factor. METHODS The mixture of cilostazol and cilostazol impurity Ⅰ was dissolved in deuterated dimethyl sulfoxide. A portion of the solution was determined by qNMR, while the other portion of the solution was diluted with water-acetonitrile (60:40) and analyzed by HPLC. The correction factor of cilostazol impurity Ⅰ was calculated with the response signals from qNMR and the peak areas from HPLC. The correction factor of cilostazol impurity Ⅰ was also determined by HPLC standard curve method. A mixed solution containing residual solvent was prepared to simulate the effect of solvents in determining correction factors. When the content of cilostazol impurity Ⅰ was inaccurately assigned due to residual solvent, difference between qNMR-HPLC method and standard curve method was compared. RESULTS When the contents of cilostazol and cilostazol impurity Ⅰ were assigned accurately, the correction factors for cilostazol impurity Ⅰ by qNMR-HPLC method and HPLC standard curve method were 1.74 and 1.76, respectively, which were basically consistent with the pharmacopoeial results. When cilostazol impurity Ⅰ contained residual solvents, and there was an error in the content assignment, the correction factor of the determination by the qNMR-HPLC technique was still 1.72, and the result was not affected by the accuracy of the content. While the correction factor of HPLC standard curve was 2.01, which was deviated from the actual results. CONCLUSION Compared with the HPLC standard curve method, the qNMR-HPLC coupling technique is independent of the accuracy of the content of the substance to be measured and the weighing volume, and does not require purification to prepare a high purity compound. qNMR-HPLC is a powerful tool in the determination of impurity correction factors.

OBJECTIVE To investigate the efficacy and safety of different preparations of intravenous immunoglobulin (IVIG) for Kawasaki disease treatment in children. METHODS A retrospective analysis of medical records of pediatric Kawasaki disease patients at our institution in recent three years was conducted. Patients were divided into preparation A group and preparation B group based on the type of IVIG preparations used. Both groups received IVIG (2 g·kg^-1) in conjunction with aspirin therapy following Kawasaki disease diagnosis. Changes in body temperature, white blood cell count, platelet count, C-reactive protein levels, IVIG cost, length of hospital stay, and adverse events were compared. RESULTS According to the drug labeling, there are certain differences between preparation A and preparation B. Preparation B exhibited a significantly shorter duration of fever resolution compared with preparation A (P<0.005), and at the first temperature test after infusion, the body temperature of preparation B group was significantly lower than that of preparation A group (P<0.05). Furthermore, preparation B demonstrated a more significant reduction in white blood cell count following treatment (P=0.010). In terms of hospital stay duration and IVIG monotherapy costs, preparation B had significantly shorter hospital stay (P<0.05) and lower per-unit IVIG expense (P<0.001). No significant differences were observed between the two groups in terms of adverse events, including rashes, infusion interruptions due to high fever, liver injury, and coronary dilation. CONCLUSION There is significant difference in efficacy between different preparations of intravenous immunoglobulin in the treatment of Kawasaki disease in children, but the safety is similar.