Objective: To determine the concentration of vanillic acid by polyoxometal-based host-guest frame material modified glass carbon electrode. Methods: A stable polyacid-based host-guest metal-organic framework material PMoV@MIL-100(Fe) was synthesized by combining vanadium-substituted phosphomolybdic acid (PMoV) with MIL-100(Fe) according to the synthesis method of polyacid-based host-guest framework materials. Functional composite material PMoV@MIL-100(Fe)@Pt was prepared by composite metal nanoparticles with PMoV@MIL-100(Fe) by solution method, and was used to modify glassy carbon electrode, detect vanillic acid, and prepare vanillic acid electrochemical sensor. Results: Under optimal conditions, the vanillic acid electrochemical sensor showed wide linear range and high sensitivity. Rapid and sensitive detection of vanillic acid could be performed under mild conditions with excellent stability. Conclusion: The electrochemical sensor constructed in this experiment provides a new idea for the detection of vanillic acid.

Objective: To establish a method for determination of aconite alkaloids in Fengshi Gutong tablets. Methods: Analysis was performed on a WatersXSelect^® CSH C₁₈（250 nm×4.6 mm, 5 μm） column with mobile phase consisting of 0.1% acetic acid aqueous solution, acetonitrile and methanol with gradient elution at a flow rate of 1.0 mL·min^-1. The detection wavelength was set at 245nm and the column temperature was 30 ℃. Results: Twelve aconite alkaloids could be separated well. When the injection amounts of ranaconitine, benzoylmesaconine, benzoylmesaconine, benzoylmesaconine, acoforestinine and beiwutine, mesaconitine, hypaconitine, indaconitine, aconitine, yunaconitine, bulleyaconitine A were 0.022 8-0.136 6 μg, 0.041 5-0.249 0 μg, 0.033 5-0.200 8 μg, 0.033 9-0.203 1 μg, 0.033 1-0.198 6 μg, 0.040 3-0.241 6 μg, 0.030 2-0.181 4 μg, 0.028 6-0.171 6 μg, 0.033 6-0.201 6 μg, 0.030 3-0.181 9 μg, 0.063 4-0.381 5 μg and 0.034 0-0.204 2 μg, respectively, the peak area showed a good linear relationship with the injection amounts. The average recovery rates of 12 aconite alkaloid components ranged from 93.6% to 101.5%, and the RSD ranged from 0.55% to 2.6%. The average contents of ranaconitine, benzoylmesaconine, benzoylmesaconine, benzoylmesaconine, acoforestinine and beiwutine, mesaconitine, hypaconitine, indaconitine and yunaconitine in three batches of Fengshi Gutong tablets (batch No. 211227, 220823 and 230425) were 0.492 μg, 65.78 μg, 7.319 μg, 10.164 μg, 1.068 μg, 5.583 μg, 2.573 μg, 5.865 μg, 2.021 μg, 2.050 μg, respectively. And aconitine and bulleyaconitine A were not detected. Conclusions: The established method is accurate and reliable, and can be used for the determination of 12 aconite alkaloids compounds in Fengshi Gutong tablets to achieve comprehensive quality control of preparations.

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 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 the HPLC fingerprint of Suhexiang pills and determine the contents of gallic acid, piperine, costunolide, eugenol and cinnamic acid simultaneously, and to provide reference for the quality control method of Suhexiang pills. Methods: The separation was performed on a Swell Chromplus C₁₈ column (250 mm×4.6 mm, 5 μm) with methanol (A)-0.05% phosphoric acid solution (B) as the mobile phase at a flow rate of 1.0 mL·min^-1. The detection wavelength was 220 nm and the column temperature was 30 ℃. Fifteen batches of Suhexiang pills were analyzed. The “Traditional Chinese Medicine Fingerprint Similarity Evalution System” was used to establish the reference fingerprint and the similarity analysis was evaluated in combination with cluster analysis and principal component analysis. Results: The fingerprint of Suhexiang pills was established，and the similarities of 15 batches of samples were above 0.9. Thirteen common peaks were identified，and five components including galliic acid，cinnamic acid，eugenol，piperine and costunolide were identified. The contents of the five components were 2.506-3.652 mg·g^-1，0.666-0.876 mg·g^-1，3.834-5.140 mg·g^-1，0.884-1.306 mg·g^-1 and 19.908-55.704 mg·g^-1，respectively. Using cluster analysis，15 batches of Suhexiang pills were divided into four categories. Three principal components were selected，and the cumulative variance contribution rate was 76.102%，indicating that the principal components could contain most of the information of the original data. Conclusion: The fingerprint and multi-component content determination method of Suhexiang pills are stable and reliable，which can provide reference for the quality control and clinical application of Suhexiang pills.

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 investigate the source and quality of commercially available Epimedium based on HPLC fingerprint and multi-index assay, so as to provide reference for its quality evaluation and resource development. Methods: Forty batches of commercially available epimedium were collected. Fingerprint analysis and determination of the total flavonoid glycosides were performed by HPLC method. The total flavonoid glycosides were determined by UV method. And the contents of their extracts were determined. Stoichiometric analysis was employed to perform the comprehensive quality evaluation of different medicinal materials of Epimedium. Results: There were 9 different plant sources of Epimedium in the market. The five authentic Epimedium were of good quality, and the local varieties were coarse Epimedium. The contents were high in Epimedium from Hunan, while the contents were low in Epimedium from Qianling and Sichuan. The comprehensive quality ranking of Epimedium indicated that Epimedium Fletchum and Epimedium Fletchum ranked first by the principal component analysis method. The ranking of the same variety varied greatly with different habitats, and the Qianling Epimedium and adulterated Epimedium ranked last. The results of cluster analysis and principal component analysis were basically consistent. The cluster analysis showed distinguishing significance in plant sources, habitats, processing methods and contents. Based on the common pattern of E. brevicornu, a total of 8 common peaks were identified for 9 species of E. Brevicornu, and their similarities ranged from 0.066 to 0.979. Differences were observed among different species. Conclusion: Both the fingerprint similarity and the chemical model of content determination can be used to evaluate the species and quality of Epimedium.

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 an HPLC method for the simultaneous determination of six components (uridine, adenine, adenosine (R, S)-goitrine, guanosine, clemastanin B) in Radix Isatidis, and to investigate the linear calibration with two reference substances (LCTRS) method for the qualitative analysis of multiple components in Radix Isatidis. Methods: HPLC method was used, with methanol as mobile phase A and water as mobile phase B. Gradient elution (0-3 min, 3% A; 3-18 min, 3% A→14% A; 18-25 min, 14% A→26% A; 25-34 min, 26% A; 34-40 min, 26% A→46% A; 40-60 min, 46% A→90% A) was performed at a flow rate of 0.8 mL·min^-1. The column temperature was 30 ℃, the detection wavelengths were 254 nm (0-32 min) and 230 nm (32-60 min). The injection volume was 10 μL. The actual retention time of 6 components in Radix Isatidis was determined on 13 C₁₈ chromatographic columns of different brands and models. Guanosine and clemastanin B were used as double reference compounds, and LCTRS method was used to locate the chromatographic peak of each component. Three unknown chromatographic columns were used for method validation. Using guanosine as a reference substance, the relative retention time method was used to predict the retention time of the other 5 components. The predictive accuracy and column coincidence of these two methods were compared. Results: The LCTRS method could effectively predict and qualitatively analyze the retention time of six indicator components. Compared with the relative retention time method, the LCTRS method had higher accuracy in predicting results and better column universality. Conclusion: The LCTRS method for simultaneous determination of multiple components in Radix Isatidis is feasible and accurate, with simple operation and good durability, and has promotional value.