To evaluate the differences of aqueous extract protein content and kinds among Hudilong origins in order to provide reference for the identification and quality evaluation.

The protein content of aqueous extract was determined, and the differences among the origins were compared by nonparametric tests. The protein fingerprints were established by SDS-PAGE, and the band matrix was constructed based on the molecular weight of the bands. The differences of the protein kinds among the 3 origins were compared and analyzed by CA and OPLS-DA.

There was no statistically significant difference in the aqueous extract protein content between the P. vulgaris and P. guillelmi, but both of them differed significantly from that of P. pectinifera (P<0.01). Aqueous extract protein content. from P. pectinifera was higher than from P. vulgaris or P. guillelmi. The results of SDS-PAGE showed that there were differences in the molecular masses and quantities of the protein bands of the 3 origins with relative molecular masses ranging from 10×10³ to 250×10³. The results of the CA and OPLS-DA showed that each of the origin of Hudilong could be differentiated. The differences of the protein relative molecular masses were 133.7×10³, 10.4×10³, 66.9×10³, 51.9×10³, 13.4×10³, 7×10³, 29.4×10³, 14.6×10³, 62.2×10³, 81.1×10³, 89.2×10³, 7.7×10³, and 101.7×10³, respectively.

There are differences in the aqueous extract protein among 3 origins of Hudilong. The P. vulgaris and P. guillelmi have high similarity in the aqueous extract protein components content and kinds. SDS-PAGE technique could be used to provide reference for the identification and quality evaluation.

To identify the risks and provide a scientific basis for the formulation of the limit standards by the determination and risk assessment of pesticide residues in Yixintong tablets and hawthorn leaf extracts.

The samples were extracted with acetonitrile by high speed homogenater, and purified by solid - phase extraction(SPE) column of envi-carb/NH₂. 262 pesticides in hawthorn leaf extracts and Yixintong tablets were detected by GC-MS/MS and LC-MS/MS. Moreover, the acute and chronic intake risks were calculated using the deterministic risk assessment model.

25 and 31 types of pesticides were detected in hawthorn leaf extracts and Yixintong tablets, respectively. 18 types of pesticides were detected in all of them. The contents were 0.004-1.457 mg·kg^-1, 0.007-2.1 mg·kg^-1, respectively. The results of risk assessment revealed that the short-term hazard index(HIa) of hexaconazole was 1 and the risk was needed to be paid more attention. HIa and long-term hazard index(HIc) of other pesticides detected in Yixintong tablets and hawthorn leaf extracts were much lower than 1, indicating an acceptable risk.

In the present study, the pesticide residues were explored, and their health risks were assessed；therefore our study provided technical support for the improvement of risk assessment applicable to traditional Chinese medicines (TCMs) and the formulation of relevant pesticide residue limit standards.

To identify the authenticity of the Longgu, and reveal the scientific illustration for superior quality of white Longgu.

In this study, the quality of 88 batches of Longgu was evaluated by cluster analysis, principal component analysis and correlation analysis based on character, ultraviolet fluorescence and results obtained with inductively coupled plasma mass spectrometry, color analyzer and nuclear radiation detector.

The Longgu spots on the surface of Longgu were the key identification points of genuine Longgu, and the Longgu spots were mainly composed of Mn elements. Bright blue fluorescence could be seen in the fresh section of keel under 365 nm. Among the 42 elements determined, only Mg and Zn were higher in the fake Longgu than in the genuine Longgu. Other elements were lower in the fake Longgu than in the genuine Longgu. Heavy metals and harmful elements in counterfeit samples were below limits in pharmacopoeia, and cluster analysis showed that all counterfeit samples were grouped into one group separately. The Longgu could be divided into four color types: white, cyan, yellow and brown. The larger a * was, the redder the color of Longgu was, the higher the contents of Fe and harmful elements (As, Cu, Pb and Cd) were. And the contents of element V were strongly correlated with those of As, Cu, Pb and U, which could be used as index element of heavy metals and harmful elements. The contents of radioactive element U and the radiation values were highest in brown Longgu, second highest in yellow Longgu was the, lower in cyan Longgu, and lowest in white Longgu. The average content of U in brown Longgu was 0.021% with the highest content of 0.09%, and the average radiation value was 0.33 μSv·h^-1 (Background value was 0.18 μSv·h^-1) with the highest value of 0.47 μSv·h^-1. The average content of U in yellow Longgu was 0.012% with the highest content of 0.021%, and the average radiation value was 0.32 μSv·h^-1 with the highest value of 0.39 μSv·h^-1. The average content of U in cyan Longgu was 0.008 9% with the highest content of 0.012%, and the average radiation value was 0.30 μSv·h^-1 with the highest value of 0.35 μSv·h^-1. The average content of U in white Longgu was 0.006 7% with the highest content of 0.009 1%, and the average radiation value was 0.28 μSv·h^-1 with the highest value of 0.32 μSv·h^-1.

Considering the harmful elements and radiation safety, the white Longgu has better quality, and the brown Longgu has worse quality.

To establish an HPLC-CAD method for the determination of related substances in peramivir injection and to analyze the impurity profile.

Welch Ultimate Polar RP column (250 mm×4.6 mm, 3.5 μm) was used with 5 mmol·L^-1 ammonium formate solution (pH adjusted to 4.6 with formic acid) -acetonitrile (95∶5) as mobile phase A and mobile phase A-acetonitrile (50∶50) as mobile phase B under gradient elution at the flow rate of 0.8 mL·min^-1. Column temperature was 30 ℃. CAD atomizer temperature was 50 ℃, acquisition frequency was 5 Hz and filter was 3.6 s. The degradation pathway and impurity structure of peramivir were estimated by LC-MS/MS.

The separation between peramivir and the impurities was good. The mass concentration of peramivir and 7 impurities showed good linear relationships with the peak area in the ranges of 0.2-12 μg·mL^-1. The limits of quantitation were 4.32-12.8 ng, and the limits of detection were 2.16-6.44 ng. The average recovery (n=9) of impurity Ⅲ was 96.2%, RSD was 1.8%. The control solution and the test product solution were stable for 102 hours at room temperature. After fine-tuning the parameters of liquid chromatography, there was no influence on the detection results of related substances. Based on LC-MS/MS, the structure and possible degradation pathway of the degraded related substances were deduced.

This method is accurate, reliable and specific for the determination of related substances in peramivir injection, and provides a basis for the establishment of quality standards for this variety.

To investigate the metabolites and metabolic pathways of WJ-14, a novel anti-depressant compound, in rats and human liver microsomes by HPLC-MS/MS.

Samples of rat plasma and urine, as well as in vitro incubation system of human liver microsomes were analyzed by HPLC-MS/MS. Chromatographic conditions: the chromatographic conditions were as follows: Hypersil BDS C₁₈ (100 mm×4.6 mm, 3 μm) column, with 0.1% formic acid aqueous solution (A)-methanol (B) as the mobile phase, gradient elution flow rate 0.4 mL·min^-1, column temperature 40 ℃, and injection volume 5 μL. The identification of metabolites and speculation on the metabolic pathways of WJ-14 were conducted by comparing the fragmentation mode and characteristic fragment ions of WJ-14.

A total of 12 metabolites of WJ-14 were identified, including 6 in rat and 6 in human liver microsomes, with two metabolites were detected in both rat and human liver microsomes. The major metabolic pathways of WJ-14 were identified as hydroxylation, demethylation, hydration, glucuronidation and methylation.

In this work, we investigated the metabolites and metabolic pathways of WJ-14 using HPLC-MS/MS for the first time. The metabolism of WJ-14 occurred in phase Ⅰ and phase Ⅱ metabolic reactions, in which hydroxylation in phase Ⅰ is dominant. The results of the current study may provide valuable information for the formulation development and application of WJ-14.

To establish a quality evaluation method of Erdi Xiaoke mixture based on HPLC fingerprint, multi-component determination and chemometric analysis.

The separation was performed on a 30 ℃ thermostatic Acutfex PW-C₁₈ (250 mm×4.6 mm, 5 μm) column, with the mobile phase comprising of acetonitrile -0.1% phosphoric acid flowing at 1.0 mL·min^-1 in a gradient elution manner, and the detection wavelength was set at 285 nm to establish HPLC fingerprint of Erdi Xiaoke mixture. Contents of 11 components [5-hydroxymethyl furfural(5-HMF), salvianic acid A, chlorogenic acid, caffeic acid, calycosin-7-glucoside, acteoside, isochlorogenic acid B, isochlorogenic acid C, rosmarinic acid, salvianolic acid B and formononetin] in 12 batches of Erdi Xiaoke mixture were determined, and similarity evaluation, cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to comprehensively evaluate the quality of Erdi Xiaoke mixture.

The 11 constituents showed good linear relationships within their respective ranges(r≥0.999 6), with average recovery rates of 95.3%-101.5%, and the RSDs were in the range of 1.1%-2.3%. There were 18 common peaks in the fingerprints for 12 batches of samples with the similarities above 0.960. The range of the content of the above 11 components in the 12 batches of samples were as follows as 43.40-65.70, 219.43-274.70, 119.00-164.78, 11.87-18.17, 42.80-56.86, 10.01-20.71, 8.21-14.73, 40.15-57.39, 43.29-60.29, 755.22-1 208.50, 2.47-4.44 μg·mL^-1. And 12 batches of Erdi Xiaoke mixture could be clustered into three categories with some differences in sample quality among different batches. In addition, 5-HMF, isochlorogenic acid B and chlorogenic acid were the main potential markers affecting the quality of Erdi Xiaoke mixture.

This method is stable, reliable, accurate and sensitive, and can be used to control and comprehensively evaluate the quality of Erdi Xiaoke mixture.

To establish an UPLC-MS method for the analysis of salidroside, methyloleoside, specnuezhenide, acteoside, oleuropein, and G13 in plasma, and the differences in pharmacokinetic profiles of five different concoctions of Ligustri Lucidi Fructus, wine Ligustri Lucidi Fructus, vinegar Ligustri Lucidi Fructus, salt Ligustri Lucidi Fructus and steamed Ligustri Lucidi Fructus in vivo of rats were investigated.

SPF-grade male SD rats were randomly divided into 6 groups, and were given 4.2 g·kg^-1 of aqueous extracts of different concoctions of Ligustri Lucidi Fructus by gavage (for the amount of raw drug), and the plasma samples were methanol-precipitated proteins with geniposide as the internal standard, and the plasma samples were used to determine salidroside, methyloleoside, specnuezhenide, acteoside, oleuropein, and G13 in the plasma of the rats in the different time points by using the negative-ion SIM mode of UPLC-MS/MS. Kinetica 5.1 software was used to calculate the pharmacokinetic parameters, and GraphPad Prism 8.4.0 software was used to analyze the data.

The mass concentrations of salidroside, methyloleoside, specnuezhenide, acteoside and oleuropein were in the range of 2.00-1 385 ng·mL^-1, and the mass concentration of G13 was in the range of 1.30-650 ng·mL^-1, with good linearity, the relative standard deviations of the precision were all less than 10%, the recoveries of the extracts were all in the range of 85%-105%, and the matrix effect and stability were in accordance with the requirements of biological samples. The results of pharmacokinetic study showed that the AUC_0-∞, MRT_0-t, t_1/2, and C_max of acteoside were significantly higher in wine Ligustri Lucidi Fructus (P<0.01) compared to raw Ligustri Lucidi Fructus, which was 751.36 ng·mL^-1·h, 5.87 h, 377.82 h, and 38.11 ng·mL^-1, respectively. C_max, AUC_0-∞, and t_1/2 were significantly higher (P<0.01) for specnuezhenide and G13 in salt Ligustri Lucidi Fructus compared to raw Ligustri Lucidi Fructus, with C_max of specnuezhenide and G13 in salt Ligustri Lucidi Fructus were 66.45 ng·mL^-1 and 204.27 ng·mL^-1, respectively. The AUC_0-∞ were 342.69 ng·mL^-1 and 423.44 ng·mL^-1·h, and t_1/2 were 101.64 h and 15.98 h, respectively. Compared to raw Ligustri Lucidi Fructus, the C_max of oleuropein was significantly higher (P<0.05) in vinegar Ligustri Lucidi Fructus, wine Ligustri Lucidi Fructus, and steamed Ligustri Lucidi Fructus with 66.81 ng·mL^-1, 68.00 ng·mL^-1, and 66.38 ng·mL^-1, respectively. The AUC_0-∞ of salidroside and methyloleoside was the highest in vinegar Ligustri Lucidi Fructus, which was 5 782.74 ng·mL^-1 and 545.26 ng·mL^-1·h, respectively.

This study reveals the changing law of the six active ingredients in different artillery products of Ligustri Lucidi Fructus in vivo and their different characteristics, which provides a basis for the clinical application of different artillery products of Ligustri Lucidi Fructus.

To study the related substances in salbutamol sulfate inhalation aerosol during national drug sampling and testing, and to compare the impurity content and evaluate the quality between samples from various enterprises.

HPLC external standard method was utilized to simultaneously determine the content of related substances A, B, C, D, E, F, G, H, I, J, N, and other unknown impurities in salbutamol sulfate inhalation aerosol. Thermo Synchronis C₈ chromatography column (250 mm×4.6 mm, 5 μm) was used. Sodium heptane sulfonate solution-acetonitrile was used as the mobile phase. Linear gradient elution was performed and the flow rate was 1.0 mL·min^-1. Column temperature was 40 ℃ and detection wavelength was 220 nm. Injection volume was 20 μL. The sources of impurities through forced degradation experiments were explored. Toxicity prediction software for impurity toxicity assessment was applied.

After method validation, the specificity of the method was good, and the separation degree between each impurity peak was greater than 1.5. RSDs of precision test were 0.30%-1.7%(n=6)；mass concentrations of linear range were from 0.050 to 5.000 μg·mL^-1(r=0.999 9). Limit of quantitative was in the range of 0.025-0.200 μg·mL^-1, limit of detection was in the range of 0.008-0.070 μg·mL^-1. The repeatability RSD of raw materials was 0.80%-3.8%, and the recovery rate was 95.2%-104.8%. The repeatability RSD of inhaled aerosol was 1.2%-2.9%, and the recovery rate was 98.7%-102.8%. The forced degradation test showed that impurities D, F, I, J, and N were all degradation impurities. 110 batches of samples were checked and the results of the relevant substances met the regulations. In the samples of diverse enterprises, impurities C, D, F, and N were detected more frequently, while impurities E, G, and H were not detected. Impurity J was only detected in one batch. The predicted impurity D by QSAR software falls to ICH M7 (R2) level 2.

The established method is sensitive and accurate, and can accurately quantify the content of related substances in salbutamol sulfate aerosol, providing effective technical support for systematic supervision. Further toxicity studies should be conducted on impurity D and reasonable limits should be established.

To study the dynamic changes of puerarin and icariin in plasma and hippocampus of mice，to observe the changes of depression-related protein expression in hippocampus，and to explore the pharmacokinetic changes of Shenge Bushen capsules (SBC) and its correlation with the changes of hippocampus-related protein expression.

Puerarin and icariin in plasma and hippocampus of normal mice were analyzed by UPLC-MS/MS-ABSCIEX QTRAP 5500 triple quadrupole series linear ion trap mass spectrometry method using Waters Acquity HSS T3 (50 mm×2.1 mm，1.8 μm) column with 0.05% formic acid water-acetonitrile-methanol (1:1) containing 0.05% formic acid as mobile phases gradient elution at a flow rate of 0.2 mL·min^-1 and electrospray ion source under negative ion model. The proteins in hippocampus were expressed by Western blot assay.

Puerarin and icariin were detected in plasma and hippocampus after the oral administration，including plasma puerarin t_1/2 2.45 h，icariin t_1/2 3.59 h，hippocampus puerarin t_1/2 4.37 h and icariin t_1/2 8.5 h. The plasma concentration of puerarin accounts for 27.3% and that of icariin accounts for 1.34% of the dosage taken. Puerarin in hippocampus accounts for 2.47% of puerarin absorbed into blood，while icariin in hippocampus accounts for 73.56% of icariin absorbed into blood. The expressions of restrictive silencing factor (NRSF)，brain-derived neurotrophic factor (BDNF) and its downstream protein tyrosine kinase B (TrkB)，solute carrier transporter 6a4 (SLC6A4)，glucocorticoid receptor (GR) and μ opioid receptor (MOR) in hippocampus were all up-regulated after SBC administration，in which icariin was negatively correlated with NRSF expression，puerarin was positively correlated with BDNF-TrkB and MOR，and GR had no obvious correlation with the proteins.

Puerarin and icariin can be absorbed into the blood and distributed in the hippocampus after oral administration of SBC in mice. Icariin was easier to pass through the blood-brain barrier and distribute in the hippocampus than puerarin. The expression of hippocampus-related proteins was related to the changes of puerarin and icariin concentrations，suggesting that the targets of the two components as well as SBC were related to these proteins. This study provides an important experimental basis for the pharmacokinetic study of SBC and also provides a material basis of its antidepressant effect.