Objective: To explore the fragmentation patterns of synthetic cannabinoids by electron impact (EI) ionization mass spectrometry. Methods: Forty synthetic cannabinoids were systematically investigated by gas chromatography coupled to mass spectrometry (GC-MS). Ionization mode was EI (70 eV) and the acquisition range was m/z 50-600. Results: According to the different structures of the “head group” and “linked group”, forty synthetic cannabinoids were divided into six categories, namely cumyl-carboxamide type, adamantyl-carboxamide type, carbamoyl/methyl butyrate-carboxamide type, naphthylformyl type, benzoyl/phenylacetyl type and tetramethylcyclopropane-acyl type. Through the analysis of the mass spectrum of synthetic cannabinoids, the fragmentation pathways and characteristic ions of different types of synthetic cannabinoids were given. The main EI-MS fragmentation patterns of synthetic cannabinoids were that both sides of the carbonyl group in the “linking group” undergo α-cleavage, and the N atom on the indole/indazole parent nucleus was prone to γ-H rearrangement, and loss of a R₁. In addition, fragment ions m/z 116, 130, 144 and fragment ions m/z 117, 131, 145 were the characteristic fragments of indazole and indole parent nucleus, which could be used to identify the parent nucleus of synthetic cannabinoids. Conclusion: These kind of compounds have strong fragmentation regularity. When standard substances are lacking or commercial mass spectral libraries are difficult to obtain, the proposed synthetic cannabinoids EI-MS fragmentation pathways can help to rapidly identify the structures of unknown synthetic cannabinoids.

Objective: To analyze the fractions and relative contents of volatile oils of Magnoliae Flos at different harvesting periods, to elucidate the dynamic pattern of changes in the chemical composition of Magnoliae Flos at five harvesting periods, and to evaluate its antioxidant and antimicrobial activities. Methods: The volatile oils of Magnoliae Flos at five harvesting periods was extracted by water vapour distillation, and the chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS) technique, and the relative content of each constituent was calculated. The constituents of Magnoliae Flos at the five harvesting periods were analyzed by PLS-DA analysis, which was used in combination with the VIP value to screen out the differential compounds. The antioxidant activity of the volatile oil of Magnoliae Flos was determined by ferric ion reducing antioxidant power (FRAP) method, and its in vitro antimicrobial activity was investigated by 96-well plate method. Results: The total volatile oils content of Magnoliae Flos was the highest in samples at the 4th harvesting period (10 February 2023). Thirty-eight components were identified in the volatile oils of Magnoliae Flos, and 12 differential compounds were screened, including γ-muurolene, elemene, δ-cadinene and α-terpineol, etc. The relative contents of γ-muurolene, alloaeromadendrene, borneol, camphor and cis-4-thujanol were the largest in samples at the 4th harvesting period, which was basically in line with the trend of the change of volatile oil content. The volatile oils in samples at five harvesting period showed certain antioxidant and antibacterial activities. And that in samples at the 4th harvesting period showed the strongest antioxidant activity and the inhibition ability against all five species of bacteria. Conclusion: The chemical composition of the volatile oils in Magnoliae Flos was basically the same in in samples at five harvesting periods, but there is a significant difference in the relative content of its volatile components in each harvesting period, and it is presumed that the beginning of February is the optimal harvesting period for Magnoliae Flos.

Objective: To investigate the flavor components of Epimedium extract and analyze pyrolysis products of the extracts from Epimedium. Methods: Ultra fast gas phase electronic nose was adopted to analyze volatile components in ethanol extracts of Epimedium and the alcohol extracting process of Epimedium was optimized by orthogonal experiment. Epimedium extract was pyrolysed to simulate cigarette smoking by TG-GC-MS. The lysates of Epimedium extract were analysed in a nitrogen environment, and the possible lysate mechanism of the products was reasonably speculated. Results: 22 volatile components were detected in the ethanol extracts of Epimedium at different concentrations. Concentration of Epimedium extracted with 60% ethanol was superior than others. Analyzing pyrolysis products of Epimedium extract, 78 compounds were identified at 150, 300 and 450 ℃, including aldehydes, ketones, alcohols, phenols, furans and benzene series. Conclusion: Ethanol extraction of Epimedium contains many aromatic volatile components. A large number of ketones, alcohols, phenols, furans and other volatile aroma compounds are produced after pyrolysis of Epimedium extract.

Objective: To establish a liquid-liquid extraction liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of the concentration of glycyrrhizic acid and glycyrrhetinic acid in human plasma. Methods: Glycyrrhizic acid, glycyrrhetinic acid and the internal standard Apixaban-¹³C-d₃(IS) were added to 0.1 mL of human blank plasma. 50 μL of ionization reagent (20% formic acid solution), 490 μL of ethyl acetate and 210 μL of methyl tert-butyl ether were used as extractant. The supernatant was dried by nitrogen, and the residue was dissolved with 200 μL acetonitrile-water (1∶1) containing 0.2% formic acid. And 5 μL of resulting solution was injected to the LC-MS/MS for analysis. Chromatographic conditions: the saparation was performed on a Boston Μni C₁₈(50 mm×2.1 mm,3 μm)column with mobile phase consisting of 0.2% formic acid aqueous solution(mobile phase A)and 0.2% formic acid acetonitrile solution(mobile phase B)by gradient elution. The flow rate was 0.6 mL·min^-1, the temperature of column was 40 ℃. The sample volume was 5 μL, and the temperature of the sampler was 4 ℃. Mass spectrometry conditions: multiple reaction montoring(MRM) was performed on a triple quadrupole mass spectrometer equipped with a ESI source in the positive mode. The detection ion pairs were m/z 823.4→453.3(glycyrrhizic acid), m/z 471.4→189.0 (glycyrrhetinic acid)and m/z 464.3→447.1(IS) respectively. Results: The calibration curves were linear over the concentrion ranges of 0.5-80 ng·mL^-1 for glycyrrhizic acid and 2-800 ng·mL^-1 for glycyrrhetinic acid (r>0.99) in the plasma, the lower limits of quantifications (LLOQ) were 0.5 ng·mL^-1(glycyrrhizic acid) and 2 ng·mL^-1(glycyrrhetinic acid), respectively. Inter-and intra-batch precisions (RSDs) were less than 6.8%, and the accuracy ranged from 92.3% to 104.2%. The recovery rates of glycyrrhizic acid and glycyrrhetinic acid were about 28.0% and 40.0% separately, and the recoveries of IS were about 65.0%, the precision (RSDs) were less than 7.9%. The normalized matrix factors of glycyrrhizic acid and glycyrrhetinic acid were about 1, and the precision (RSDs) were less than 7.3%. Conclusion: The method is sensitive, accurate, simple, rapid and applicable to simultaneous determination of the concentration of glycyrrhizic acid and glycyrrhetinic acid in human plasma.

Objective: To establish a dual derivatization method combined with tandem mass spectrometry for the simultaneous determination of 18 different steroid hormones in human serum. Methods: Serum samples were treated with hydroxylamine and 1,2-dimethylimidazole-5-sulfonyl chloride for derivatization, and the resulting compounds were analyzed by liquid chromatography-tandem mass spectrometry in positive ion selected reaction monitoring (SRM) mode. The Kinetex^®C₈ column (100 mm×2.1 mm, 2.6 μm) was used for the separation. Mobile phase A (0.1% acetic acid in water) and mobile phase B (0.1% acetic acid in methanol) with gradient elution(0-0.5 min,35%B;0.5-5 min, 35%B→100%B; 5.0-5.1 min,100%B→35%B; 5.1-7 min, 35%B)at the flow rate of 0.4 mL·min^-1 were applied. Column temperature was set at 40 ℃. Injection volume was 20 μL and collection time was 6 min. Results: The linearity correlation coefficients for all 18 steroid hormones were greater than 0.99, the recovery rates ranged from 85% to 115%, and the precision RSD was less than 15%. This method was successfully applied to the analysis of serum samples from 156 healthy subjects (75 males and 61 females), and reference intervals were established. Conclusion: This method can be used to simultaneously determine 18 types of steroid hormones, such as pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, corticosterone, cortisol, 11-deoxycorticosterone, 21-deoxycorticosterone, aldosterone, testosterone, androstanedione, dehydroepiandrosterone, sulfated dehydroepiandrosterone, adrenocorticotropin, 18-hydroxicorticotropin, estrone, estradiol, and estriol using non-solid-phase extraction.

Objective: To develop a method to assess bioactivity of trasfer factor (TF) based on their protective effcets on Jurkat E6-1 cells. Methods: Proliferative effects of different concentrations of TF on 6-mercaptopurine(6-MP) treated Jurkat E6-1 cells detected by CCK-8 assay and the precision was validated. Results: TF exhibited protective activity to change the inhibitory concentration (IC₅₀) value of 6-MP from(0.46±0.10)μg·mL^-1 to (1.11±0.30)μg·mL^-1 when treated with 20 μg·mL^-1 TF. When cells were treated with 6-MP,there is a good linear relationship between the concentration of TF and cell survival rate, r>0.95. At each concentration level of the dose-response curve, the RSD was less than 20%. The median effective concentration (EC₅₀) of TF was (28.49±9.60)μg·mL^-1, and the confidence limit was less than 20%. Conclusions: TF significantly improved Jurkat E6-1 cell survival rate in dose dependent manner when challenged with 6-MP. It may be suitable for evaluate bioactivity of TF.

Objective: To establish an authentication method for Eupolyphaga sinensis based on loop-mediated isothermal amplification (LAMP). Methods: Two pairs of primers (outer primer DB-F3,DB-B3 and inner primer DB-FIP,DB-BIP) were designed according to Eupolyphaga sinensis COⅠspecific loci. The reaction system containing template, primers, Bst DNA polymerase and methyl red-phenol red indicator amplified at 63 ℃ for 90 min. The LAMP results were determined by visual observation and compared with DNA barcoding and polymerase chain reaction (PCR) methods to investigate the specificity and sensitivity. Results: All 11 batches of the authentic Eupolyphaga sinensis sample tubes colour changed from purple red to orange yellow, while sample tubes of 1 batch of Steleophaga plancyi, 9 batches Opisthoplatia orientalis and 1 batch of the adulterant, Cybister tripunctatus orientalis, remained purple red after the reaction. The LAMP results were consistent with those of DNA barcoding and PCR. The detection limit of LAMP was 0.984 ng·mL^-1. Conclusion: The established LAMP method is accurate, sensitive and easy to operate, low equipment required, and can be applied to the rapid screening of the authentication of Eupolyphaga sinensis.

Objective: To study the difference of chemical composition between ancient and modern clinical prescriptions of Baihe Dihuang decoction by multi-component analysis. Methods: The polysaccharide extract of Baihe Dihuang decoction was determined by anthrone-sulfuric acid-ultraviolet spectrophotometer under 580 nm ultraviolet light. Six monosaccharides and oligosaccharides in the ancient and modern Baihe Dihuang decoction were determined by UPLC-CAD. An Agilent InfinityLab Poroshell 120 HILIC-Z (3.0×100 mm,2.7 μm) column was used. The mobile phase was consisted of A 0.2% ammonia in acetonitrile and 0.2% ammonia in water with gradient elution. The flow rate was 0.8 mL·min^-1, the column temperature 35 ℃ and the CAD detector nebulizer temperature was 70 ℃. Data acquisition frequency was 10 Hz and the filter was 1.05 s. The injection volumn was 2 μL. The non-polar components were qualitatively and quantitatively analyzed by high performance liquid-quadrupole tandem time-of-flight mass spectrometry (UPLC-Q TOF MS/MS) and high performance liquid-phase tandem triple quadrupole mass spectrometry (UPLC-QQQ MS), respectively. The differential components of ancient and modern Baihe Dihuang Decoction were analyzed and 9 components were determined. Results: The linear relationship of 6 mono-oligosaccharides/polysaccharides and 9 differential components was good (r>0.999). The average recoveries ranged from 97.7% to 104.9% with RSDs≤3.1%. Precision, repeatability and stability met the requirements. The results showed that the conents of all the analytes in ancient and modern Baihe Dihuang decoction were significantly different (P<0.05). Conclusion: The methods used in this experiment are accurate, sensitive, stable and reproducible, which can provide reference for the quality control and pharmacodynamic material basic research of Baihe Dihuang Decoction.

Objective: To analyze the difference of chemical constituents in rhizomes and leaves of Polygonatum sibiricum from Shaanxi Province. Methods: Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Orbitrap HRMS) was developed to determine the chemical constituents in rhizomes and leaves of Polygonatum sibiricum from Shaanxi Province. The data were analyzed by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The structures of chemical markers in rhizomes and leaves of Polygonatum sibiricum were identified based on accurate primary mass spectrometry and secondary mass spectrometry fragment ion, combined with the reference map, software database searching and related literature. Results: A total of 45 compounds were identified, there were 26 chemical ingredients with significant differences distinguished by the method of OPLS-DA, including 10 amino acids, 6 flavonoids, 3 organic acids, 2 saccharides, 1coumarin and 4 alkaloids. Conclusion: The chemical markers of amino acids, organic acids and alkaloids are mainly distributed in rhizomes, and the chemical markers of flavonoids are mainly concentrated in leaves part of the plant. It is suggested that the potential multiple utilization of rhizomes and leaves of Polygonatum sibiricum from Shaanxi Province.

Objective: To establish methods of thin-layer chromatography(TLC) qualitative identification and quantitative analysis of multi-components by single-marker (QAMS) according to the characterization results of the main components of Iris tectorum Maxim.. Methods: Firstly, the main components of Iris tectorum were characterized by ultra performance liquid chromatography tandem time-of-flight mass spectrometry(UPLC-Q TOF MS/MS) technology,using an ACQUITY UPLC^® BEH C₁₈ column (100 mm×2.1 mm, 1.7 μm) with gradient elution using 0.1% formic acid-water (A)-acetonitrile (B) as mobile phase, and the data were collected in positive ion mode. In addition, the qualitative identification method was established by the TLC. Finally, quantitative analysis was performed on an Agilent 5 TC-C₁₈ column (250 mm×4.6 mm, 5 μm). The mobile phase was 0.05% phosphoric acid water(A)-acetonitrile(B) with gradient elution. The QAMS method was established by using tectorigenin as an internal standard to establish its relative correction factor with tectoridin and irigenin, and the contents of three isoflavones were calculated by the correction factor. At the same time, the accuracy and feasibility of the QAMS method was verified by the external standard method(ESM). Results: Five isoflavones were characterized (tectoridin, iristectorin B, tectorigenin, irigenin and iristectorigenin A) by UPLC-Q TOF MS/MS. A TLC identification method for four components, tectoridin, iristectorin B, tectorigenin and irigenin was established. In this study, the linearity of the three components of tectoridin, tectorigenin and irigenin were good in a certain concentration range (r>0.999), with the average recoveries of 97.7%-100.8% and the RSDs of 1.1%-2.9%, and the relative correction factors of tectoridin and irigenin were 1.114 5 and 0.827 0, respectively. And the repeatability of the correction factors was good under different experimental conditions (RSD<5%). There was no significant difference between the contents of the three analytes in ten different batches of Iris tectorum determined by QAMS and ESM. The contents of tectorigenin, tectoridin and irigenin in 10 batches of Iris tectorum from different habitats determined by QAMS were not significantly different from those determined by ESM. Conclusion: Based on the characterized components of Iris tectorum medicinal materials, the TLC and QAMS methods with high-content and stable ingredients are established. The methods are simple and stable, which can be used for qualitative and quantitative analysis and quality evaluation of Iris tectorum.

Objective: To establish an integrated model of biosensing technology for the study of zebrafish, and to study and verify the interaction between cetirizine hydrochloride, fexofenadine hydrochloride, mizolastine, and ebastine, with the key target of allergic rhinitis, macrophage migration inhibitory factor(MIF). Methods: A MIF-high electron mobility transistor(HEMT) biosensor was constructed using AlGaAs/GaAs HEMT as the sensing element and MIF as the biological element. The interaction strength between cetirizine hydrochloride, fexofenadine hydrochloride, mizolastine, and ebastine and MIF was characterized. Furthermore, a zebrafish immune inflammation model was established. The number of neutrophil migration in the blank control group, model group, positive drug group, and low, medium, and high-dose drug groups were recorded. And the drug inflammation inhibition rate were calculated. Results: Cetirizine hydrochloride exhibited the strongest binding ability to MIF, with a dissociation constant of 7.05×10^-13 mol·L^-1. The K_D for the interactions between fexofenadine hydrochloride, mizolastine, and ebastine with MIF were 2.34×10^-8 mol·L^-1, 1.94×10^-7 mol·L^-1 and 2.44×10^-8 mol·L^-1, respectively. All of them had binding effects with MIF. Further the validation using the zebrafish immune anti-inflammatory model revealed that all four antihistamines significantly reduced the number of neutrophil migrations, among which 100 μmol·L^-1 of cetirizine hydrochloride achieved a neutrophil migration inhibition rate of 68.5%. Conclusion: This study explores the interaction between antihistamines and the key protein target of AR through the integration of biosensing technology and zebrafish experiments. It was found that MIF is a potential target for antihistamine drugs to exert their pharmacological effects. It provides an important reference for research on drug efficacy and its association with targets.

Objective: To analyze the peptide components of bran-fried Bombyx batryticatus decoction and virtually screen anti-epileptic peptides with GAT-1 binding activity. Methods: The peptide compounds in bran-fried Bombyx batryticatus decoction were extensively identified by micro-flow liquid chromatography-tandem mass spectrometry(μLC-MS/MS) technology. Potential anti-epileptic peptides were screened through bioinformatics and in silico simulations, including bioactivity prediction, blood-brain barrier permeability prediction, toxicity prediction, and molecular docking. Finally, molecular dynamics simulations were employed to analyze the interactions between peptides and the target protein. Results: A total of 384 peptides were identified from bran-fried Bombyx batryticatus decoction, and 19 peptides with higher binding affinities to GAT-1 than that of the control drug were screened as potential anti-epileptic peptides. The FDHFDFDAF exhibited the highest binding affinity, followed by the EHYAWGIK. Two peptides primarily interacted with GAT-1 through hydrogen bonds, hydrophobic, and electrostatic interactions. Molecular dynamics simulations confirmed the binding stability of FDHFDFDAF and EHYAWGIK to GAT-1. Conclusion: Peptidomics and in silico virtual screening techniques facilitate the rapid discovery of active peptide components in animal-derived traditional Chinese medicine. This study has significant reference value for researching anti-epileptic peptides in bran-fried Bombyx batryticatus decoction, and provides new insights for the study of peptide components in animal-derived traditional Chinese medicine.

Objective: To study the whole chain microbial load in the production of a traditional Chinese medicine pill. Methods: A type of traditional Chinese medicine pills prescription was selected as the research object, including the medicinal materials and their purified medicinal materials, intermediates (mixed powder, waiting for inner packaging pills), and finished products. The microbial limit test method was established according to the 2020 edition of the Chinese Pharmacopoeia(ChP), to detect and analyze the microbial loads of all samples collected throughout the entire chain from medicinal materials to finished products of the traditional Chinese medicine pills. Results: The microbial content of different medicinal materials varies greatly, and the varieties with higer risk factors for microbial contamination in medicinal materials were Perillae Folium, Pogostemonis Herba, and Angelicae Dahuricae Radix. The production process of Perillae Folium and Pogostemonis Herba needed to be improved and adjusted from “medicinal material to pure medicinal material”. There was a positive correlation between the microbial load of medicinal materials and intermediate and finished products. Conclusion: The study on the microbial load throughout the entire production chain of traditional Chinese patent medicines can help us to understand the transmission of microorganisms at each stage. This knowledge is crucial for identifying weak links in the production process and the key areas for process control, and has an important guiding significance for traditional Chinese patent medicines manufacturers to improve and explore their production process.

Objective: To classify the microbiological examination methods of live microecological products according to the results of suitability test for the enumeration method, and establish the decision-making method for microbiological examination of live microecological products. Methods: The suitability test of the microbiological examination method of live microecological products was performed according to the requirements of the General Chapter 1105 in the Chinese Pharmacopoeia Vol Ⅳ. Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger were used as the test strains. Tryptic Soy Agar, Sabouraud Dextrose Agar, Rose Bengal Agar and Rose Bengal Agar Ⅱ were used as medium. Results: For live microecological products composed of anaerobic bacteria or microorganisms with growth characteristics significantly different from the test strains, the suitability test for the method could be performed according to General Chapter 1105. For live microecological products containing Bacillus and enterococcus, the suitability test for the fungal enumeration method could be performed according to General Chapter 1105. Live microecological products could not be tested for suitability of the method due to the influence of the component microorganisms, gram-negative bacteria could be controlled using selective medium. In addition, some potentially harmful microorganisms could be controlled based on risk assessment according to the route of administration and the population of drug users. Conclusion: This study provides a basis for the selection of the microbiological examination methods for live microecological products, and further complements and improves the quality standard system of live microecological products.

Objective: To compare the differences of the volatile oil components of Pimpinella thellungiana Wolff extracted with different methods by GC-MS combined with retention index. Methods: Volatile oil of Pimpinella thellungiana Wolff were extracted using steam distillation, salting-out assisted steam distillation and enzymatic hydrolysis-assisted steam distillation. The chemical components of volatile oil extracted by different methods were analyzed by GC-MS combined with retention index, while the relative contents of volatile oil components were calculated by peak area normalization method. Results: Among the three extraction methods, the extraction rate of volatile oil was as follows, enzymatic hydrolysis-assisted steam distillation > salting-out assisted steam distillation ≈ steam distillation method. The most comprehensive types of volatile oil were extracted by salting-out assisted steam distillation, followed by enzymatic hydrolysis-assisted steam distillation, and finally by steam distillation. The main types of the volatile oil components of Pimpinella thellungiana Wolff extracted with different methods were basically unchanged, but the relative contents of various compounds were different. A total of 47 compounds were identified from the volatile oil extracted with the three methods, including 35 common compounds. The main chemical structural types of these compounds were terpenes, terpenes, terpenoids, aromatics and aliphatics, among which the higher content compounds were β-bisabolene(17.25%-19.42%), caryophyllene oxide (12.90%-15.70%), 1-(3-methyl-2-butenoxy)-4-(1-propenyl)benzene (5.02%-9.36%) and β-pinene (5.31%-6.62%). Conclusion: The chemical composition types and relative contents of the volatile oil of Pimpinella thellungiana Wolff extracted with different methods are different, but the differences are not significant. The results can provide reference for the selection of suitable extraction methods and further utilization of volatile oil from Pimpinella thellungiana Wolff.

Objective: To establish a method for determination of potential anti-vascular disease active components of Semen raphani based on cell membrane chromatographic (CMC) model of human umbilical vein cell (HUVEC). Methods: The HUVEC cell membrane chromatography coupled with HPLC-ESI IT TOF MS system were used to screen the active components of Semen raphani. The selected active components were further applied to ox-LDL induced HUVEC to verify their protective effects. Results: Two retained components were selected from Semen raphani by this method. One component was identified as erucinic acid by comparing with the reference material. Compared with the model group, the cell survival rates of the erucinic acid pretreatment groups increased significantly. The amount of ICAM-1 and VCAM-1 decreased in a dose-dependent manner. Bcl-2 protein levels decreased and Bax protein levels increased, with statistical significance (P<0.05). Conclusion: The method can rapidly obtain active ingredients from complex traditional Chinese medicines. It provides a reference for the application of cell membrane chromatography and the development of Semen raphani.

Objective: To establish and optimize a rapid direct analysis of real time tandem mass spectrometry (DART-MS/MS) method for the rapid detection of 12 benzodiazepines in blood and urine that can be used in forensic toxicology work. Methods: A DART ion source was used in conjunction with an API4000 Q Trap mass spectrometer. A DART 12Dip-It^TM autosampling module with a module travel speed of 0.6 mm·s^-1, a sample volume of 5 μL, and a gate voltage of 200 V were applied. The mass spectrometry section scans in positive ion mode using multiple reaction monitoring (MRM) mode. After further optimization, the DART-MS/MS method was validated and applied to real case samples. Results: Ethyl acetate was selected as the extractant for liquid-liquid extraction and the temperature of the carrier gas heater was optimized. The method has good selectivity and does not interfere with delayed effects. The linearity was good, and the limits of detection (LODs) for the targets in blood and urine were in the ranges of 0.5-10 ng·mL^-1 and 0.2-2 ng·mL^-1, respectively. And the limits of quantification (LOQs) were in the ranges of 1-50 ng·mL^-1 and 0.5-5 ng·mL^-1, respectively. The recoveries ranged from 78.8% to 119%, and the matrix effects ranged from -17.5% to 18.5%. The intra- and inter-day precisions were not greater than 14.4% for the high and intermediate concentrations, and not greater than 18.1% at the limits of quantification. This method enables fast and accurate examination of case samples. Conclusion: The method is fast and convenient, with good sensitivity, and can be applied to the research and work of rapid detection of toxicants to improve the efficiency of detection.

Objective: To establish an HPLC-MS/MS method for the determination of FCN-437c in human plasma and its application to the phase I clinical study of FCN-437c. Methods: Following protein precipitation, plasma was injected and measured using HPLC-MS/MS method. The analytes were separated on a YMC Triart PFP column (50 mm×2.1 mm, 5 μm) using 0.5% formic acid (containing 5 mmol·L^-1 of ammonium acetate, A) and acetonitrile (B) as the mobile phase with gradient elution at the flow rate of 0.5 mL·min^-1, the column temperature was set at 35 ℃, the injection amount was 2 μL, and the injector temperature was 10 ℃. MS detection was performed with multiple reaction monitoring (MRM) mode using positive electrospray ionization. The ion transitions were m/z 549.4→449.4 for FCN-437c and m/z 552.3→449.3 for FCN-437-D3, respectively. Other mass spectrometry parameters were TEM, 500 ℃,GS1, 276 kPa,GS2, 207 kPa,DP, 100 V,CXP, 25 V. Results: The linear range of FCN-437c in human plasma was 5-1 000 ng·mL^-1 (r=0.999 0). The lower limit of quantification was 5 ng·mL^-1. The intra-batch and inter-batch precisions were less than 2.0% and 4.1%, respectively. The average recovery was 104.0% (FCN-437c), 78.6% (FCN-437-D3), and the internal standard normalized matrix factor was 100%-102%. The stock solution of FCN-437c was stable at 4 ℃ for 202 d, the working solution of FCN-437c and internal standard were stable at room temperature for 24 h. FCN-437c in human plasma was investigated to be stable at room temperature for 20 h, four cycles of freeze-thaw, -20 ℃ for 134 d, -80 ℃ for 662 d, as well as for 24 h in the autosampler after treatment. The whole blood samples were stable at room temperature for 4 h. This method was applied to the determination FCN-437c in human plasma, and the deviation between the test results and the initial values of 97.0% ISR samples was within ±20%. The accuracy was 102.0%-108.0% after 10-fold dilution of plasma samples. The cumulative ratio of R_AUC0-24 and R_Cmax was 1.33 times and 1.59 times when FCN-437c was administered continuously compared with single administration. Conclusion: This method is simple, accurate, robust and specific, which can meet the requirements of quantitative analysis of FCN-437c in human plasma and also can be used to determine FCN-473c in human plasma of the phase I clinical study.

Objective: To establish a UPLC-MS/MS method for the simultaneous determination of 47 pesticide residues in Sucrose. Methods: The residues in the sample were extracted by acidic acetonitrile, concentrated by blow dry with nitrogen gas and then analyzed by using LC-MS/MS with multiple reaction monitoring(MRM). Blank matrix standard curve with internal standard method was used to determine the residue contents. Results: Each substance to be tested had good linearity relationship within certain concentration (r>0.995). The limits of quantification (LOQ) were 0.01-2.00 μg·kg^-1. The recoveries of 47 pesticide residues at three levels of 10, 25 and 50 μg·kg^-1 were from 71.0% to 106.0% with the RSD of 0.90%-8.5% in sucrose. 2 batches in 15 batches of sucrose samples were detected thiamethoxam and thiamethoxam, while the rest were not detected. Conclusion: The method is simple, rapid and characterized with acceptable sensitivity and accuracy to meet the requirements of the determination of analysis, and this developed procedure is suitable for the determination of pesticide residues in Sucrose, can be used for quality control of sucrose.

Objective: To establish a liquid chromatography mass spectrometry (LC-MS) method for the detection of nepasaikosaponin K in Bupleurum marginatum var. stenophyllum, which could be used to quickly identify whether Bupleurum marginatum var. stenophyllum was substituted for Bupleurum or was mixed with Bupleurum. At the same time, nepasaikosaponin K was detected in 16 batches of Bupleurum marginatum var. stenophyllum and 157 batches of bupleurum decoction pieces. Methods: Ultra-high performance liquid chromatography (UPLC) coupled with triple quadruple mass spectrometry (QqQ MS) was used to detect nepasaikosaponin K in Bupleurum and Bupleurum marginatum var. stenophyllum. Determination was carried out with the application of a Phenomenex Kinetex C₁₈ (2.1 mm×100 mm, 1.7 μm) column at temperature of 25 ℃. The mobile phase was composed of acetonitrile(A) and water(B) with gradient elution(0-5 min 15%A→25%A;5-0 min,25%A→30%A; 30-35 min,30%A→90%A;35-36 min,90%A→15%A;36-40 min,15%A)at a flow rate of 0.3 mL·min^-1. The electrospray ionization(ESI^-) source was performed in multiple reaction monitoring(MRM)mode of the transitions of m/z 943.5→797.3, m/z 943.5→635.3, and m/z 943.5→781.3. Results: The contents of nepasaikosaponin K detected in 16 batches of Bupleurum marginatum var. stenophyllum were much higher than minimum requirements. Nepasaikosaponin K was detected in 8 out of 157 batches of Bupleurum. Conclusion: The established method is verified by methodology to be specific, so as to develop a supplementary test method for the components of Bupleurum and Bupleurum marginatum var. stenophyllum. to improve the quality control standard and authentic identification investigation, and to effectively control the quality of Bupleurum decoction pieces and guarantee its clinical efficacy.

Objective: To determine the in vitro release degree of musk sustained-release mini-tablets by reciprocating cylinder method. Methods: Gas chromatography was used to establish the in vitro analysis of muscone, which is the main active ingredient of sustained-release mini-tablets. Comparing its equilibrium solubility in different concentrations of Tween-80, hexadecyl trimethyl ammonium bromide (CTAB) and sodium dodecyl sulfate (SDS), to determine the type of release medium. The effects of residence time, drip time, reciprocation rate, and screen mesh sizes on the release behaviours of muscone were investigated. Results were compared with that of the paddle method. Meanwhile, a preliminary investigation into the mechanism of its release was also carried out. Results: The linearity of the constructed analytical method was better, the RSD values of precision, repeatability and stability were less than 3%, and the spiked sample recovery was qualified. The release medium for the reciprocating cartridge method was 200 mL of 0.05% Tween-80, with a dwell and drip time of 20 s, a reciprocation rate of 15 dip·min^-1, and upper and lower screen mesh sizes of 20 and 40 respectively. Compared with the paddle method, Muscone was completely released under different pH conditions in the reciprocating cylinder method, and the release curve increased smoothly. Conclusion: The study can provide some references for the reciprocating cylinder method in the in vitro evaluation of sustained-release prescriptions of traditional Chinese medicine.

Objective: To develop a method for characterizing in vitro release of cyclosporine ophthalmic emulsion. Methods: The Franz diffusion cell and the polyvinylidene fluoride membrane were adopted with buffer-ethanol (40∶60) as receiving media. The sampling time was set at 60,125,190,255,320,385 min, respectively. Results: The in vitro release method showed that the inertia of membrane, specificity, sensitivity and selectivity met the requirements. The validation of HPLC showed that the quantitative limit of the method was 0.07 μg·mL^-1 and a good linear relationship between the concentration range of 0.07-44.62 μg·mL^-1. The average recovery was 98.9%. Compared with the original preparation by FDA guideline, the in vitro release of the self-developed preparation was the same as the reference preparation. Conclusions: This method is suitable for the in vitro release evaluation of cyclosporine eye drops.

Thyroid-stimulating hormone (TSH) is a glycoprotein hormone produced by the anterior pituitary. It can regulate the synthesis and secretion of thyroid hormone in thyroid follicular cells, which has important physiological significance. As a drug, it has important application value. Biological activity detection is an effective and necessary to evaluate its quality. This article discusses the signal transduction mechanism of thyroid stimulating hormone, the clinical diagnosis and treatment of thyroid stimulating hormone, and the determination method of biological activity.

Therapeutic oligonucleotides (OGNs) drugs are artificially synthesized single or double stranded short nucleic acids, typically 15 to 30 base pairs in length. OGNs have been rapidly developed as new therapeutic drugs with increasing attention in the discovery and development of drugs concerning various disease fields. Compared with Europe and America, there are currently no other OGNs drugs listed in China, except for Spinraza, which has been approved for marketing as an orphan drug. The development of OGNs in China started relatively late and is still in its early stages of development. However, the OGNs drug market in China is anticipated to grow quickly due to the country's large population, high patient demand, ongoing support for the development of oligonucleotide drugs in the future, and the steady maturation of related technologies by domestic businesses. Because of their special physicochemical characteristics, OGNs drugs are challenging to design biological analysis techniques. Currently, there are few reports on quantitative analysis methods for oligonucleotide drugs in China. Therefore, the development of sensitive and reliable bioanalysis methods for oligonucleotides is the key to investigate oligonucleotides' pharmacokinetic and pharmacodynamic properties. Liquid chromatography-mass spectrometry (LC-MS) can quantify OGNs and their metabolites concurrently, compared with traditional ELISA approaches. Numerous benefits come with using LC-MS, in particular, the extensive use of high-resolution mass spectrometry allows for the identification of metabolites, which provides details on base composition and sequence structure, in addition to quantitative information about target oligonucleotides. It has now emerged as the go-to technique for OGN quantitative analysis. The application of LC-MS in the identification of therapeutic oligonucleotide medicines is the primary focus of this paper, which also discusses its benefits and drawbacks. Lastly, it looks at the LC-MS development trend for oligonucleotide detection, which includes a lower detection level and potential general methods.

Objective: To establish a fingerprint of Gualou Guizhi decoction and a method for determination of multi-component to clarify the transfer rule of quantities and quality from decoction pieces and substance benchmarks. Methods: Fifteen batches of Gualou Guizhi decoction substance benchmarks were prepared and analyzed by the method of HPLC. Traditional Chinese medicine (TCM) Chromatographic Fingerprint Similarity Evaluation Software (2012) and hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) was used to evaluate the quality differences between batches of substance benchmarks, and screen the main chemical compositions that were led to quality discrepancy. The main differential components were determined by HPLC. And their contents, yields and transfer rates were analyzed for transfer rule of quantities and quality. Results: HPLC fingerprint of Gualou Guizhi decoction was established. A total of 30 common peaks in the fingerprint of Gualou Guizhi decoction substance benchmarks were assigned. Compared with the reference standards, 9 components were identified as gallic acid, albiflorin, paeoniflorin, liquiritin, ononin, liquiritigenin, cinnamic acid, isoliquiritigenin and glycyrrhizic acid. The similarities of 15 batches of Gualou Guizhi decoction substance benchmarks samples were all above 0.920. All of them were divided into two categories by three chemical recognition modes for 5 major differential components, including glycyrrhizic acid, liquiritigenin, paeoniflorin, iquiritin, and ononin. The HPLC was also applied to determine the contents of multi-components and the method validation results were good. The average transfer rates of five index components were 51.65%, 40.46%, 74.74%, 60.06%, and 34.54%, respectively and their average extraction rate was 14.20%. Conclusion: The critical quality properties of Gualou Guizhi decoction can be stably transferred from decoction pieces to substance benchmarks. The method of fingerprint and content determination is accurate and reliable, which is able to provide technique for quality control of Gualou Guizhi decoction formulation.

Objective: To establish high-performance liquid chromatography method using cyclodextrin as a mobile phase additive for the determination of ursolic acid and oleanolic acid in Corni Fructus, providing a basis for quality control of Corni Fructus. Methods: Molecular design and response surface method were used to optimize the determination method of ursolic acid and oleanolic acid in Corni Fructus. Agilent Eclipse XDB C₁₈ column (150 mm×4.6 mm, 5 μm) was adopted as the stationary phase, the mobile phase was 3 mmol·L^-1 γ-cyclodextrin solution (contained 0.05% phosphoric acid)-acetonitrile (60∶40), the flow rate was 1.0 mL·min^-1, the column temperature was 25 ℃ and the detecting wavelength was 210 nm. Results: The resolution of chromatographic peaks of ursolic acid and oleanolic acid was 3.81. Linearities were good in the range of 400-4 000 ng for ursolic acid and in the range of 200-2 000 ng for oleanolic acid. The RSDs for precision, reproducibility, and stability were less than 1.4%, 1.8%, and 1.7%, respectively. The recovery rates were 98.5% (RSD=1.7%) and 99.1% (RSD=2.0%). The contents of ursolic acid in three batches of Corni Fructus were 2.66 mg·g^-1, 2.35 mg·g^-1 and 2.91 mg·g^-1, and those of oleanolic acid were 0.83 mg·g^-1, 0.78 mg·g^-1and 1.12 mg·g^-1, respectively. Conclusion: This method is simple, accurate, reliable, and environmental friendly, and can be used as a quality control for Corni Fructus.

Objective: To establish the UPLC-MS/MS method for simultaneous determination of 9 components (nicotinic acid, kaempferol, swertisin, quercetin, luteolin, rutin, vitexin, spinosin, salicylic acid) in Desmodium caudatum (Thunb.) DC. and construct a back propagation(BP) neural network model to predict the origin of Desmodium caudatum (Thunb.) DC. from different habitats. Methods: The chromatographic separation was achieved on an Agilent Zorbax SB C₁₈ column (50 mm×3.0 mm,1.8 μm). The mobile phase consisted of methanol-0.1% acctic acid (containing 0.02 mol·L^-1 ammonium acetate) at a flow rate of 0.3 mL·min^-1 with gradient elution, the MS analysis were performed by multiple reaction monitoring (MRM) under ESI⁺ and ESI. A correlation analysis was conducted on the contents of each component, and a BP neural network model was constructed to distinguish Desmodium caudatum (Thunb.) DC. from different habitats. Results: Under the optimized conditions, 9 components(nicotinic acid, kaempferol, swertisin, quercetin, luteolin, rutin, vitexin, spinosin, salicylic acid) showed good linear relationships in the ranges of 0.388 8-38.88 ng·mL^-1, 10.07-1 006.6 ng·mL^-1, 34.22-34 221.6 ng·mL^-1, 3.944-394.4 ng·mL^-1, 2.124-212.4 ng·mL^-1, 4.344-434.4 ng·mL^-1, 46.50-4 650.1 ng·mL^-1, 1.649-164.9 ng·mL^-1, 4.880-488.0 ng·mL^-1, respectively (r>0.995 1), whose average recoveries were 96.9%-103.9% (RSDs<1.9%). The contents of the above nine components in 40 batches of Desmodium caudatum (Thunb.) DC. were 1.657-7.407 μg·g^-1, 15.801-64.488 μg·g^-1, 1 068.348-4 270.780 μg·g^-1, 10.608-123.228 μg·g^-1, 3.897-16.802 μg·g^-1, 1.269-97.834 μg·g^-1, 405.285-1 955.796 μg·g^-1, 13.614-36.124 μg·g^-1, 4.417-87.509 μg·g^-1, respectively. According to correlation analysis, four components (swertisin, rutin, spinosin, and luteolin) in Desmodium caudatum (Thunb.) DC. showed a highly linear positive correlation, indicating that these four components had a certain synergistic effect in Desmodium caudatum (Thunb.) DC.. The BP neural network model was constructed to predict Desmodium caudatum (Thunb.) DC. from different habitats, and the accuracy of the test set reached 92.3%. Conclusion: The method is simple, sensitive and efficient, and can be used for the rapid determination of the components in Desmodium caudatum (Thunb.) DC.. Using the BP neural network model to predict the habitats plays a significant role in tracing the origin of Desmodium caudatum (Thunb.) DC..

Objective: To establish an UPLC quantitative analysis method for the simultaneous determination of 13 components in Yankening tablets, including phellodendrine, coptisine, baicalin, palmatine, berberine, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, chrysophanol, physcion. Methods: Agilent Eclipse Plus C₁₈(100 mm×2.1 mm, 1.8 μm) column was used with acetonitrile-0.1% phosphoric acid as mobile phase, and gradient elution at a flow rate of 0.3 mL·min^-1. The detection wavelengths were 210 nm and 254 nm. The column temperature was 40 ℃. Results: Phellodendrine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, berberine hydrochloride, baicalin, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, chrysophanol, physcion showed good linear relationships within their concentration range of 0.97-48.63 μg·mL^-1, 0.95-47.52 μg·mL^-1, 0.86-43.15 μg·mL^-1, 0.86-43.19 μg·mL^-1, 0.89-44.37 μg·mL^-1, 1.00-49.84 μg·mL^-1, 1.02-51.01 μg·mL^-1, 0.97-48.31 μg·mL^-1, 0.99-49.50 μg·mL^-1, 1.04-51.80 μg·mL^-1, 1.00-50.04 μg·mL^-1, 1.00-49.80 μg·mL^-1, 1.01-50.64 μg·mL^-1. The average recoveries(n=6) were 95.2%, 96.7%, 95.9%, 98.3%, 94.1%,97.6%, 99.2%, 96.6%, 95.5%, 97.2%, 97.0%, 97.8%, 98.7%, RSD values were all less than 2.0%. The contents of the 13 chemical components in 3 batches of samples were 1.367-1.488 mg·g^-1(calculated as phellodendrine hydrochloride), 0.378-0.412 mg·g^-1(calculated as coptisine hydrochloride), 4.611-5.505 mg·g^-1, 0.324-0.407 mg·g^-1(calculated as palmatine hydrochloride), 3.665-3.878 mg·g^-1(calculated as berberine hydrochloride), 1.107-1.682 mg·g^-1, 0.392-0.941 mg·g^-1, 0.076-0.105 mg·g^-1, 0.097-0.116 mg·g^-1, 1.059-1.213 mg·g^-1, 0.149-0.167 mg·g^-1, 0.213-0.239 mg·g^-1, 0.047-0.059 mg·g^-1. Conclusion: The method is accurate, high analysis efficiency, good repeatability, it can be used to control the quality of Yankening tablets.

Objective: To establish a solid-phase extraction-high performance liquid chromatography(SPE-HPLC) method for the rapid detection of related substances of vitamin D₃, trans vitamin D₃ (impurity A), 7-dehydrocholesterol (impurity B), lumisterol 3 (impurity C), and isotachysterol 3 (impurity D), in vitamin D drops (soft capsules). Methods: The content, which was approximately equivalent to vitamin D₃ 1 548 IU, was taken in about 1.2 g, weighed accurately, put into a test tube, 4 mL of isooctane was added to it, and it was swirled and mixed well. Purification was performed using solid phase extraction columns, with n-hexane-ethyl acetate(85∶15) as the elution agent and anhydrous ethanol as the desorption agent. A Luna Silica(2) 100 Å silica gel column(150 mm×4.6 mm, 3 μm) was used, with n-hexane-n-pentanol (996.5∶3.5) as the mobile phase. The content of vitamin D₃ impurities was calculated using the adding standard calibration factors principal component self-control method. Results: The separation degree between pre-vitamin D₃ and vegetable oil was greater than 1.5. The linear range of vitamin D₃ was 0.02-0.80 μg·mL^-1, with r=0.999 5. The linear range of impurity A was 0.02 -0.80 μg·mL^-1, with r=0.999 9. The linear range of impurity B was 0.02-0.80 μg·mL^-1, with r=0.999 9. The linear range of impurity C was 0.02-0.80 μg·mL^-1, with r=0.999 9. The linear range of impurity D was 0.02-0.80 μg·mL^-1, with r=0.999 9. The average recovery rate of impurities A-D (n=9) was 93.2%-102.9%. The detection results of 3 batches of vitamin D drops (soft capsules) showed that the content of known impurities and other maximum single impurities were less than 0.5%, and the total content of impurities was less than 1.0%. Conclusion: The results indicate that the established method is suitable for the detection of vitamin D₃ related substances in vitamin D drops (soft capsules). It has the advantages of being easy to operate, providing rapid and accurate results, and providing technical assurance for the quality control of fat-soluble vitamin D₃ preparations.

Gas chromatography-Orbitrap mass spectrometry (GC-Orbitrap/MS), a developing gas chromatography-high resolution mass spectrometry approach, allows for high throughput qualitative and quantitative analysis of volatile and semi-volatile components. It has the advantages of high sensitivity, selectivity, and a wide linear dynamic range, which makes it well suitable for the analysis of a wide range of trace substances in complex matrices. In recent years, this technology has been applied in environmental science, industry, food analysis, pharmaceutical analysis, forensic science, clinical medicine and other fields. This paper presents the first review of GC-Orbitrap/MS, which not only describes the basic principles and technical characteristics, but also introduces the progress of the technique in food and pharmaceutical research. Applications in food analysis include the inspection of pesticide residues, detection of persistent organic compounds and analysis of flavor substances. In pharmaceutics, the analysis of chemical impurities and quality evaluation of traditional Chinese medicine are introduced. It is noteworthy that this technique is particularly advantageous for the identification of unknown compounds and the determination of ultra-trace components. Lastly but importantly, this review summarizes the challenges encountered in the current development of this technique, including the establishment of high-resolution standard databases, the selection and optimization of sample pre-treatment method and the application of GC-Orbitrap/MS in the field of traditional traditional Chinese medicine. A few solutions are also proposed, such as the application of variable electron voltage technique, the combination of two-dimensional gas chromatography and electrostatic field Orbitrap mass spectrometry and the integrated analysis comprehensively using multiple scan modes. These strategies are aimed to provide more advanced and accurate solutions to food, pharmaceutical, and other relevant analysis.

Patch refers to a thin sheet flexible preparation made of raw drug and suitable material for sticking on the skin, which can produce systemic or local effects. In vitro release test (IVRT) and in vitro permeation test (IVPT) are important contents of preclinical pharmaceutical research for formulation process optimization, quality control and safety and effectiveness evaluation of patch. The experimental equipment and methods used are different, the obtained experimental samples and data are different from each other, and the accuracy and precision of the experimental data are also different. Therefore, the selection of experimental equipment and the establishment of experimental methods in the in vitro experiment of IVRT & IVPT is a problem worthy of attention. In this paper, the research status of patch was summarized, the requirements of pharmacopoeia of different countries for IVRT experiments were briefly introduced, and the differences of different methods were reviewed. For IVPT experiments that have not yet been prescribed by relevant standards, the common types of experimental equipment and experimental conditions were introduced in detail, the applicability of different equipment and the influence of main experimental conditions (temperature, stirring speed, composition of acceptor solution, selected skin, etc.) on the experimental results were summarized, and the research progress of in vivo and in vitro correlation was introduced. At the same time, the validity of the experimental data was discussed, hoping to provide a useful reference for the development and research of the in vitro experimental methodology of the patch.

Objective: To analyze the chemical constituents of Quyusanjie capsules by LC/MS, and establish a method for the determination of active ingredients in Quyusanjie capsules. Methods: Using UPLC-Q TOF MS/MS technology, the Hypersil Gold C₁₈ column(100 mm×2.1 mm,1.9 μm) was used, the mobile phase was acetonitrile(A) and 0.1% formic acid in water(B) with gradient elution, at a flow rate of 0.4 mL·min^-1, the the column temperature was 40.0 ℃, and the mass spectrometry data was collected by negative ions mode scanning. Through database matching, elemental composition and fragment structure analysis, the main chemical substances in Quyusanjie capsules were identified. HPLC was used to qualitatively analyze the chemical components of Quyusanjie capsules. The Ultimate^® AQ-C₁₈ column(250 mm×4.6 mm, 5 μm) was used, the mobile phase was acetonitrile(A)-0.1% phosphoric acid(B) with gradient elution at the flow rate of 1.0 mL·min^-1, the column temperature was 25 ℃, and the detection wavelength was 203 nm. The content of naringin, neohesperidin, notoginsenoside R₁, ginsenoside Rg₁, and ginsenoside Rb₁ in 11 different batches of Quyusanjie capsules were determined using external standard method. QAMS method was established using ginsenoside Rg₁ as the internal reference. Results: Twenty-nine compounds were identified from Quyusanjie capsule. The contents of naringin, neohesperidin, notoginsenoside R₁, ginsenoside Rg₁ and ginsenoside Rb₁ measured by external standard method were 0.484-1.097 mg·g^-1, 0.341-0.618 mg·g^-1, 1.685-2.399 mg·g^-1, 5.748-8.386 mg·g^-1, 3.868-5.898 mg·g ^-1, respectively. Measured with the QAMS method, the contents of naringin, neohesperidin, notoginsenoside R₁ and ginsenoside Rb₁ were 0.516-1.153 mg·g^-1, 0.372-0.667 mg·g^-1, 1.794-2.580 mg·g^-1, 4.373-6.690 mg·g^-1, respectively. The relative error between the calculated values of the QAMS method and the measured value of the external standard method was less than 8.9%. Conclusion: UPLC-Q TOF MS/MS method can quickly identify the chemical components of Quyusanjie capsules. The established external standard method is stable and reliable, and can be used for the quality control of Quyusanjie capsules. The method of QAMS has good feasibility and is suitable for the determination of the daily production of Quyusanjie capsules.

Objective: To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of atorvastatin, two activity-related hydroxy statin metabolites and three toxicity-related statin lactones in human plasma, and its application to the study of pharmacokinetics in healthy subjects and the analysis of concentrations in patients. Methods: After acidification, plasma samples were treated by protein precipitation. The LC separation was performed on a Zorbarx SB-C₁₈(50 mm×2.1 mm, 5 μm) column. Methanol-acetonitrile (1∶1) water-methanol-acetonitrile (9∶0.5∶0.5) containing 0.05% formic acid were used as the mobile phases for gradient elution, and the flow rate was 0.35 mL·min^-1. The electric spray ionization source, positive ion mode and multi-reaction monitoring scanning were adopted for MS detection. The m/z of each targeted analyte was 559.3→440.2 for atorvastatin, 575.1→440.3 for 2-hydroxy atorvastatin acid (2-HAT) and 4-hydroxy atorvastatin acid (4-HAT), 540.9→448.2 for atorvastatin lactone (ATL), 557.2→448.2 for 2-hydroxy atorvastatin lactone (2-HATL) and 4-hydroxy atorvastatin lactone (4-HATL), and 422.2→290.0 for the internal standard of pitavastatin. After a full method validation, the developed LC-MS/MS method was used to determine the plasma samples of healthy subjects and patients after taking atorvastatin calcium tablets, and the pharmacokinetic characteristics of atorvastatin and five metabolites were analyzed. Results: The calibration curves of atorvastatin and its metabolites presented a good linear relationship in the range of 0.1-25 nmol·L^-1. The RSD of intra-and inter-day precision and the RE of accuracy were all less than 15%, and the stability was well tolerated under different conditions. In healthy subjects after oral administration of 20 mg atorvastatin calcium tablets, the respective mean values of C_max for atorvastatin, 2-HAT, 4-HAT, ATL, 2-HATL and 4-HATL were 11.48, 4.71, 0.28, 1.71, 2.52 and 2.31 nmol·L^-1, AUC_0-∞ were 87.31, 58.79, 8.60, 28.75, 45.76, 31.49 nmol·h·L^-1, t_1/2 were 7.96, 7.93, 19.58, 8.76, 8.98 and 21.37 h. After 12 h of administration, the average blood concentrations of atorvastatin, 2-HAT, 4-HAT, ATL, 2-HATL and 4-HATL in the patient were (4.16±1.31) nmol·L^-1, (2.65±1.33) nmol·L^-1, (1.15±1.16) nmol·L^-1, (2.96±1.83) nmol·L^-1, (4.27±2.00) nmol·L^-1 and (3.70±1.74) nmol·L^-1. Conclusion: The method for the simultaneous quantitative determination of atorvastatin and five metabolites in human plasma established in this study is accurate, rapid, sensitive and stable, and can be used for clinical pharmacokinetics research and plasma drug concentration monitoring. The clinical studies revealed that toxicity related lactone metabolites have a high level of exposure in humans, which requires attention to the possible risk of side effects.

Objective: To develop a high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) method for the determination of vildagliptin in human anticoagulant plasma with ethylenediamine tetra acetic acid and apply it to the study of pharmacokinetics. Methods: ¹³C-¹⁵N-vildagliptin was used as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all components were separated by a Hypurity C₁₈ column (150 mm×2.1 mm,5 μm), using a gradient elution procedure consisting of methanol and 5 mmol·L^-1 ammonium formate at a flow rate of 0.5 mL·min^-1,and the column temperature was 40 ℃. Injection volume was just 2 μL. Positive electrospray ionization was performed using multiple reaction monitoring (MRM) with transitions of m/z 304.3→154.2 for vildagliptin and m/z 310.3→160.3 for internal standard. Specificity,standard curve, lower limit of quantification,precision,recovery,matrix effect and stability were examined. Then this method was used to determine the plasma concentration of veragliptin in healthy subjects. Results: The calibration curve of vildagliptin in human plasma was linear over the concentration range of 1.11 to 534.0 ng·mL^-1. The lower limit of quantitation was 1.11 ng·mL^-1. The intra-and inter-day precisions at four quality control levels were within 0.9%-8.5%,and the accuracy was within 99.8%-109.3%. The data of short-term stability at room temperature displayed that the accuracy percentage of LQC samples was 92.0% for 0.5 h exposure, 87.6% for 1 h exposure, 71.2% for 2 h exposure. These of LQC samples chilled on ice was 102.0% for 0.5 h exposure, 94.5% for 1 h exposure, 86.6% for 2 h exposure. These results showed a phenomenon that there was a possible degradation of vildagliptin in plasma. The results of extraction recovery and matrix effect and other stability met the requirements of biological sample analysis. The pharmacokinetic study results of 8 healthy subjects showed that t_1/2 was (1.49±0.37) h, t_max was (2.06±1.11) h, C_max was (290.94±100.36) ng·mL^-1, AUC_{0-24 h} was (1 343.46±186.89) ng·h·mL^-1, AUC_0-∞ was (1 351.31±188.79) ng·h·mL^-1. Conclusion: This method is easy to operate, has high specificity, and sensitivity. It has been successfully applied to the pharmacokinetic study of 8 healthy subjects after oral administration of 50 mg vigagliptin tablets on an empty stomach. Therefore, it can be used as a reliable detection method for human pharmacokinetic research and therapeutic drug monitoring.

Objective: To establish an LC-MS/MS method for the determination of the abundance data of dexamethasone-related metabolic enzymes and transporters in the placenta of Chinese pregnant women . Methods: A Shim-pack GISS-HP C₁₈ (100 mm×2.1 mm, 1.9 μm) chromatographic column was used, and the mobile phases were consisted of 0.2% formic acid-water (phase A) and 0.2% formic acid-acetonitrile (phase B) with gradient elution at a flow rate of 0.2 mL·min^-1. The post-column phase C consisted of 0.5% ethylene glycol-acetonitrile was added at a flow rate of 0.1 mL·min^-1, and the ESI source positive ion MRM mode was used for quantitative analysis. The established method was investigated methodologically and the expression levels of 11β hydroxysteroid dehydrogenase 1(11β-HSD1), 11β hydroxysteroid dehydrogenase 2(11β-HSD2), cytochrome P450 3A4 enzyme (CYP3A4) and P-glycoprotein protein (P-gp) were quantitatively analyzed in the placenta of Chinese pregnant women in the third trimester. Results: The established LC-MS/MS method had a linear range of 0.1-100 nmol·L^-1(r >0.999). The precision and accuracy results met the requirements of the biological sample analysis method verification in Chinese Pharmacopoeia (RSD≤15%), and the stability results showed that the samples were stability. The protein abundance of 11β-HSD2, 11β-HSD1, CYP3A4 and P-gp were (84.46±59.97) pmol·g^-1, (11.44±3.73) pmol·g^-1, (8.83±2.78) pmol·g^-1 and (7.94±4.10) pmol·g^-1, respectively. Besides, the results of the study also showed that there was no significant difference in the distribution of related metabolic enzymes and transporters in different parts of the placenta. However, there was a significant difference in the abundance of P-gp in the placenta between Chinese people (7.94±4.10) pmol·g^-1 and white people (4.41±2.46) pmol·g^-1. Conclusions: The LC-MS/MS method established in this study has high accuracy and sensitivity and is suitable for detecting the abundance values of dexamethasone-related metabolic enzymes and transporters in human placenta.

Objective: To establish a suitable method to determine the structure and source of impurities of colistimethate sodium (CMS) for drug quality control studies. Methods: Frist-dimensional system: using Acquity UPLC^® Peptide CSH C₁₈(150 mm×2.1 mm, 1.7 μm) column, the mobile phase A was phosphate buffer (7.8 g·L^-1 sodium dihydrogen phosphate, adjusted to pH 6.4 with 1 mol·L^-1 sodium hydroxide)- acetonitrile (19∶1), the mobile phase B was phosphate buffer-acetonitrile (1∶1). Gradient elution was performed at a flow rate of 0.3 mL·min^-1. The column temperature was 30 ℃. Second-dimensional system: the Acquity BEH C₁₈ column (50 mm×2.1 mm, 1.7 μm) column was used with ammonium formate(A)-acetonitrile mixture as mobile phase with gradient elution. The flow rate was 0.2 mL·min^-1. The column temperature was 40 ℃. The detection wave length was 210 nm. The ESI source was used in negative ion mode. Results: The 2D-LC-Q TOF MS method was used to infer the structure of the 55 impurities in CMS, and the main sources were polymyxin E1-I, polymyxin E1-7MOA, polymyxin E3 and polymyxin E6. Conclusion: The structure and source of impurities in CMS are determined by 2D-LC-Q TOF MS, and the changes in the content of impurities such as manufacturers and production processes are evaluated, which is conducive to improving the production process and controlling drug quality at the source.

CircRNAs are a large class of endogenous single-stranded RNAs that are different from other linear RNAs, which are produced by back-splicing and fusion of either exons, introns, or both exon-intron into covalently closed loops. They are widely expressed in highly differentiated eukaryotes, and are closely related to various development and metabolic disease processes of organisms. They are characterized by stable structure, resistant to RNA degradation, conservation, and tissue-specific expression, making them ideal biomarkers for diagnosis and prognosis. Traditional methods such as Northern blotting, qRT-PCR and microarray analysis provide useful information, however, they are subject to their own shortcomings. Traditional methods are restricted in large-scale promotion in clinical trials. In recent years, in order to solve these problems, some new detection methods have emerged. In this article, we reviewed the relevant progress of all current circRNA detection methods, expounded their advantages and limitations, and discussed the challenges and future development directions.

Lonicera japonica is a kind of medicinal plant with a long history of medicinal and edible homology, which is widely distributed and has significant pharmacological activity. L. japonica contains abundant phenolic acids, flavonoids, iridoid, triterpenoid saponins, volatile oils and other active ingredients, which have antioxidant, antibacterial, antiviral and other pharmacological activities. Through consulting multiple literature databases such as Jihn.com, Wanfang and X-mol, the main literature in the past five years was mainly cited. The main active components in L. japonica, rattan and leaves and the pharmacological activities of L. japonica extract were summarized, which provided reference for the comprehensive exploitation and deep processing of L. japonica.

Objective: To comprehensively evaluate the quality of Poriae Cutis, and to establish a dual wavelength switching HPLC method for comparing the characteristic spectra of Poriae Cutis and studying the content of 11 triterpenoid components, to provide reference for the qualitative and quantitative research of Poriae Cutis. Methods: Agilent 5 HC-C₁₈(2) column(250 mm×4.6 mm, 5 μm) was adopted. Acetonitrile solution (contain 3% tetrahydrofuran) (A) and 0.1% formic acid aqueous solution (B) were used as the mobile phase with gradient elution at a flow rate of 1.0 mL·min^-1. The column temperature was 30 ℃ and the injection volume was 20 μL. The detection wavelengths were 210 and 243 nm. Results: The feature profiles developed were effective in identifying the 18 shared peaks. RSD for precision, repeatability and stability (48 h) tests were all less than 3.72%(n=6). The 11 chemical components to be measured were well separated, with good linearity in the mass range examined (all r ≥ 0.999 6). The average recovery rate was 95.4%-105.5%, and the RSD was 1.0%-3.1%. The RSDs of precision, repeatability, and stability (48 h) tests were all less than or equal to 3.0%(n=6). The results of similarity analysis showed that most of the origins of Poriae Cutis were very similar to each other. The results of content determination showed that among the 11 triterpenoid constituents, poricoic acid A accounted for the highest percentage in all batches of Poriae Cutis. In addition, the content of five components, poricoic acid A, dehydrotrametenolic acid, poricoic acid B, dehydroeburicoic acid and trametenolic acid, fluctuated relatively more, while the other components fluctuated more gently. No significant geographic variation in samples from different origins. Conclusion: A method for the determination of Poriae Cutis characteristics and multi-component content was established, which laid the foundation for quality control of Poriae Cutis.