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.

Objective: To establish an ion chromatography method for determination the content of sodium caprylate in human blood albumin products. Methods: The samples were precipitated with eluent, the suspension was centrifuged and filtered, the filtrate was injected to IC, and heptanoic acid was used as the internal standard. A Dionex InPac^TM NS1 Analytical Column (250 mm×4mm, 10 μm) and a Dionex InPac^TM NG1 Guard Column (35 mm×4 mm, 10 μm) were used,the flow rate was 1.0 mL· min^-1. The conductivity detector and ASRS 300 membrane suppressor were used, and the regenerant solution was 5 mmol·L^-1 tetrabutylsodium hydroxide solution;the column temperature was 30 ℃ and the injection volume was 25 μL. Results: The resolution between the peaks of sodium caprylate and the internal standard was greater than 1.5, and the linearity of concentration of sodium caprylate was good in the range of 0.38-2.52 mmol·L^-1, r=0.999 5 (n=6). The RSD of the repeatability test was 1.1% (n=6). The average recovery was 97.4% and RSD was 1.8% (n=9). The limits of quantification and detection were 0.19 nmol and 0.09 nmol, respectively. The determination results of the content of sodium octanoate in 20 batches of human blood albumin samples from 7 enterprises at home and abroad ranged from 0.073-0.163 mmol·g^-1. Conclusion: The method established in this study is simple to operate, accurate in results, high in sensitivity and good in repeatability, can be used for the determination of sodium caprylate content in human blood albumin products and provide a method guarantee for its quality control.

Objective: To evaluate the quality of honeyed Eriobotryae Folium from different habitats and to select the best habitat of honeyed Eriobotryae Folium preferentially based on high performance liquid chromatography fingerprinting and chemical pattern recognition methods. Methods: The detection was performed on an Acclaim^TM 120A C₁₈ (250 mm×4.6 mm, 5 μm) column with the mobile phase of 0.2% aqueous phosphoric acid (A)-acetonitrile (B) in gradient elution (0-5 min, 5%B; 5-6 min, 5%B→10%B; 6-20 min, 10%B; 20-50 min, 10%B→25%B; 50-60 min, 25%B). The volume flow rate was 1.0 mL·min^-1, the detection wavelength was 327 nm, the column temperature was 30 ℃, and the injection volume was 10 μL. The fingerprint profiles of 30 batches of honeyed Eriobotryae Folium from different habitats were established, and the fingerprint profiles combined with chemical pattern recognition were used to conduct comprehensive analysis of honeyed Eriobotryae Folium from different habitats. And cluster analysis(CA), principal component analysis (PCA) and comprehensive scoring were performed on honeyed Eriobotryae Folium from different habitats. Orthogonal partial least squares-discriminant analysis(OPLS-DA) was used to screen out the differential markers of honeyed Eriobotryae Folium from different habitats, and the habitats of honeyed Eriobotryae Folium were selected based on the comprehensive scoring. Results: The fingerprint profiles of 30 batches of honeyed Eriobotryae Folium were established. Twelve common peaks were identified, and 4 peaks were identified as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and auriculoside according to the control finger. CA divided the 30 batches of Honeyed Eriobotryae Folium samples into 6 categories. By PCA, 3 principal components were extracted, with a cumulative variance contribution of 84.315%. Six differential markers were obtained according to OPLS-DA, two of which were identified as chrysoside and chlorogenic acid. The better habitats of honeyed Eriobotryae Folium were screened as Sichuan, Guangxi, Guangdong and Shaanxi according to the comprehensive score. Conclusion: Good precision, repeatability and stability results are obtained for fingerprinting and content determination. The combination of fingerprinting and chemical pattern recognition can comprehensively evaluate the quality of honeyed Eriobotryae Folium, and this method is stable and reliable, which can provide an effective reference basis for the habitat study of honeyed Eriobotryae Folium.

Objective: To improve the liquid chromatographic determination method of cefixime granules related substance. Methods: High performance liquid chromatography was used, YMC-Triart C₁₈ column (250 mm×4.6 mm, 5 μm) was selected, 0.05 mol·L^-1 ammonium formate solution (pH 4.7)-methanol was used as mobile phase, flow rate was 1 mL·min^-1, and gradient washing was carried out.The injection volume was 10 μL. The detection wavelength was 254 nm. Results: This chromatographic condition was applied to the detection of cefixime granules. The differences between this method, the pharmacopeial method and the method of USP PF 2018 were compared, and the systematic methodological verification of specificity, linearity, accuracy, precision and durability were completed. Using pharmacopeial methods, baseline separation of degradation impurities A1~A4 or impurities B1~B4 cannot be reached, and current methods cannot be used to determine polymer B and polymer D. The method proposed in this article can make the resolution between cefixime and each specific impurities meet the requirements (R ≥1.5), and can detect and quantify polymer B and polymer D at the same time, and the resolution was better than the current method. Conclusion: This method improves the separation between cefixime and impurities, more impurities is detected and can accurate quantify specific impurities. This method has high sensitivity and good repeatability, and is suitable for the quality control of cefixime.

Illicit drugs are widely distributed in complex substrates such as biology, food, environment and drugs at minor, trace and even ultra-trace levels, which may cause acute poisoning, chronic poisoning, drug abuse and other problems. The analysis of illicit drugs has always been the focus of public safety. Solid phase extraction is a commonly used pretreatment technology for the analysis of illicit drugs in complex substrates. However, when extracting trace level illicit drugs, problems such as low sample utilization rate and poor extraction sensitivity may occur, which are difficult to meet the needs of sensitive and rapid analysis in the field of public safety. To this end, nanofibers, nanoparticles and other materials with strong size advantages are used for optimization and innovation of solid phase extraction technology. Electrospinning technology is the most commonly used method for continuous and mass production of nanofibers. It has the advantages of simple process, diverse materials and controllable fiber size, and has been widely used in the field of analysis and extraction. The electrospinning technology has experienced the development from spinning with a single polymer to blending with a variety of polymers and modifying nanoparticles with functional materials. The mechanical properties, selectivity and stability of the electrospinning nanofibers prepared have also been gradually improved, broadening the application scope of this technology in the analysis of illicit drugs. At present, the application of electrospinning technology in solid phase extraction of illicit drugs is still in its infancy. This paper systematically reviews the research status of electrospinning in traditional solid phase extraction, micro-solid phase extraction and dispersed solid phase extraction, and provides suggestions for its possible future development, in order to provide reference for further research on related issues.

Objective: To establish an UPLC-MS/MS method for simultaneous determination of 11 components(caffeic acid, afzelin, gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, salidroside, eleutheroside B, ginsenoside Re, ginsenoside Rg₁, ginsenoside Rd) in Hongwushen capsules. Methods: A Shim-pack GIST C₁₈ chromatographic column(100 mm×2.1 mm, 2 μm) was used. The mobile phase was composed of acetonitrile-0.1% acetic acid-1 mmol·L^-1 ammonium acetate aqueous solution and the gradient elution was applied. The flow rate was 0.3 mL·min^-1, the column temperature was 30 ℃, and the injection volume was 1 μL. Electrospray ionization (ESI) source and multiple reaction monitoring (MRM) mode in both positive and negative were used for ion detections. Results: The linear relationship of 11 components was good in the concentration range, and the linear correlation coefficients were all above 0.999 0. The RSD values of precision were less than 3%. The repeatability and stability were good, and the RSD values were less than 5%. The average recoveries were in the range of 97.1%-101.5% with RSDs≤3.7%. The contents of caffeic acid, afzelin, gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, salidroside, eleutheroside B, ginsenoside Re, ginsenoside Rg₁ and ginsenoside Rd in 10 batches of Hongwushen capsules were 0.010-0.013 mg·g^-1, 0.000 7-0.001 4 mg·g^-1, 0.035-0.038 mg·g^-1, 0.312-0.315 mg·g^-1, 0.413-0.417 mg·g^-1, 0.411-0.416 mg·g^-1, 4.355-4.358 mg·g^-1, 0.030-0.032 mg·g^-1, 0.993-0.999 mg·g^-1, 1.120-1.124 mg·g^-1 and 2.536-2.538 mg·g^-1, respectively. Conclusions: The method is accurate, sensitive, stable and reproducible, and can be used for the quality control of Hongwushen capsules.

Objective: To investigate the metabolites of Periploca forrestii in plasma, urine, and feces of normal and adjuvant arthritis(AA) rats, and explore the effect of rheumatoid arthritis(RA) on the metabolism of active components of P. forrestii. Methods: AA rat model was made by means of Freund’s complete adjuvant. Plasma, urine and feces of normal and AA rats were analyzed by UPLC-Q-TOF-MS^E method using ACQUITY UPLC BEH C₁₈ (100 mm×2.1 mm, 1.7 μm) column with 0.01% formic acid water-0.01% formic acid acetonitrile as mobile phase gradient elution at a flow rate of 0.25 mL·min^-1, and electrospray ion source under negative ion mode. Results: Two and three prototype components, 32 and 35 metabolites were detected in normal rats and AA model rats. The metabolic pathways mainly include monocaffeoylquinic acid reduction, methylation, ring-opening cleavage, glucuronidation, dicaffeoylquinic acid reduction, methylation, acetylation, isomerization, sulfation glucuronidation, etc. Conclusion: The metabolites in plasma and urine of AA model rats are more diverse than those in normal rats, and the disease state of RA may affect the metabolic pathway of effective components of P. forrestii in the body.

Objective: To investigate and use the different constituents in Bofonis Corium distinguishing from Bufonis Venenum based on ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry (UPLC-Q-Exactive Orbitrap MS) coupled with chemometrics. Methods: UPLC analysis was performed on a Waters Atlantis T3 (150 mm×2.1 mm, 3 μm) column with gradient elution of acetonitrile and 0.1% formic acid at a flow rate of 0.3 mL·min^-1. The column temperature was set at 30 ℃. UPLC-Q-Exactive Orbitrap MS was chosen for data collection in both positive and negative ion modes. The potential differential ions were obtained by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The Xcalibur 3.0 data processing system was utilized to extract the secondary fragmentation details of the differential ions. Subsequently, the specificity of differential ions was confirmed through the multiple reaction monitoring (MRM) mode of AB SCIEX 6500⁺ triple quadrupole mass spectrometry. Identification of adulterated Bofonis Corium in Bufonis Venenum was performed by applying the ion pairs with good specificity. Results: After the research and verification, a total of five distinction pairs (m/z: 377.5→243.2, 172.1; 330.4→170.9, 127.0; 313.4→201.2, 171.0; 452.4→280.2, 298.2 and 614.7→332.4, 281.3) were discovered, and a method for the examination of Bofonis Corium in Bufonis Venenum was established and methodologically investigated by applying the information of the different ions. The constructed method had reasonable specificity and durability. Each ion pair showed an excellent linear relationship within a specific range, with correlation coefficients above 0.996 7. The RSDs of the precision test were in the range of 1.3%-4.1%. The RSDs of the repeatability test were in the range of 1.3%-3.2%. Five batches of market samples of Bufonis Venenum were determined according to the above process, and two batches of samples showed the presence of Bofonis Corium components but did not exceed the proposed 5% adulteration limit. Conclusion: In this study, a rapid method for detecting discrepancy information based on UPLC-Q-Exactive Orbitrap MS combined with chemometrics is presented, and the discrepant information is applied to establish an identification method for adulterated Bofonis Corium in Bufonis Venenum.

Objective: To perform a comprehensive analysis of extractables in rubber by pseudotargeted metabolomics. Methods: The quality control(QC) solutions and test solutions were prepared by extracting the six kinds of rubbers with water, pH 2.6 phosphate buffer, pH 9.18 phosphate buffer, and 15% ethanol solution, respectively. The QC solutions were detected by LC-30A ultra performance liquid chromatography-Q TOF-9030 mass spectrometer in ESI⁺/ESI^- mode. The mobile phases were 0.1% formic acid-water and 0.1% formic acid-acetonitrile/0.05% ammonium-water and 0.05% ammonium-5% water-methanol. The separation chromatographic column was Waters Acquity BEH C8/HSS T3. TOF MS and MS/MS DDA scanning modes were used to detect the QC solutions. The scan results of QC solution were processed with MSConvert and MRM-Ion_Pair_Finder software to obtain MRM parameters. The extracts were detected by ExionLC UPLC-Qtrap6500+ mass spectrometer in ESI⁺/ESI^- mode with the same mobile phase and column as before, and Scheduled-MRM scan mode was applied to detect the extracts. Results: The 1 399 unknown compounds and 116 known compounds were analyzed. By extraction solution, it was found that pH 9.18 phosphate buffer and 15% ethanol solution could extract more compounds, and 27 compounds with large differences in different extraction solutions were identified. By rubbers, the affinities of different rubbers were determined by 25 differences. Conclusion: In this study, the results of the extraction of six rubbers under four extracts are analyzed. There are differences between extracts under different extraction conditions. Analytical methods can be specifically developed for these key variables for quality control follow-up studies to ensure the stability and consistency of drug quality. The variables in different rubbers are also analyzed, which can be used to make a general class determination of unknown rubber samples.

Objective: To determine seven impurities in oxytocin for injection and investigate the limit values. Methods: HPLC and principal component self-control with correction factor were adopted. The determination was performed on a Waters Xbridge C₁₈ column(150 mm×4.6 mm, 5 μm). The mobile phase consisted of 0.1 mol·L^-1 dihydrogen phosphate solution (adjusted to pH 5.4)-acetonitrile (90∶10, phase A), and acetonitrile (phase B) with gradient elution at a flow rate of 1.5 mL·min^-1. The column temperature was maintained at 32 ℃, and the detection wavelength was set at 220 nm. The injection volume was 100 μL. The linear equations of oxytocin, impurities Ac-Oxy, Oxy[Glu⁴], Oxy[+Gly¹⁰], Oxy[-NH₂], Oxy[trisulfide], Oxy[cis-dimer] and Oxy[trans-dimer] were drawn. The correction factors of each impurity related to oxytocin were calculated by slope. The contents of impurities in 3 batches of oxytocin for injection were determined and compared with the results of impurity reference method. Results: The limits of quantification for seven impurities were 2.75-5.66 ng, while the detection limits were 1.38-2.83 ng. The linear ranges of seven impurities were 0.03-3.40 μg·mL^-1 with good linearity(r>0.999). The correction factors of Ac-Oxy, Oxy[Glu⁴] and Oxy[-NH₂] were 1.1, while the correction factors of Oxy[+Gly¹⁰] and Oxy[trisulfide] were 1.2 and 0.9, respectively. The correction factors of Oxy[cis-dimer] and Oxy[trans-dimer] were both 1.3. The seven impurities were determined in 3 batches of samples by principal component self-control with correction factor. The contents of impurity Ac-Oxy were 0.96%,0.93% and 1.01%, respectively. The contents of impurity Oxy[Glu⁴] were 0.07%, 0.06% and 0.08%, respectively. The contents of impurity Oxy[+Gly¹⁰] were 0.07%, 0.04% and 0.04%, respectively. The contents of impurity Oxy[-NH₂] were 0.09%, 0.05% and 0.07%, respectively. The contents of impurity Oxy[trans-dimer] were 0.27%, 0.18% and 0.22%, respectively. The maximum single impurity contents were 0.18%-0.19%, while the total impurity contents were 1.88%-2.06%. Compared the results measured by principal component self-control with correct factor method and the impurity reference method, there was no significant difference between two methods (p>0.05). Conclusion: The method is proved to be simple, repeatable and accurate for the content determination of related substances in oxytocin for injection.

Objective: To establish a method for the rapid identification of the authenticity of Gastrodiae Rhizoma herbs based on enzymatic recombinase amplification (ERA) technique. Methods: Following the principle of ERA primer design, Oligo 7.0 software was applied to screen and optimize the ERA-specific primers of Tianma based on the ITS2 genome sequences of Gastrodia elata and its common artifacts. Primer Premier 5.0 software was applied to design the specific PCR primers for the identification of Gastrodia elata, and through the optimization of ERA and PCR reaction system, the optimal reaction time for ERA was finally determined to be 17 min, the optimal reaction temperature was 40 ℃, the optimal annealing temperature for PCR was 57 ℃, and the cycle was 32 times, and the established method was verified for sensitivity and specificity, and the samples of asparagus available in the traditional Chinese medicine The sensitivity and specificity of the established method were verified, and the commercially available asparagus samples in the market were selected for testing. Results: The designed primers for the specific identification of ERA in Gastrodia elata did not cross-react with its common forgeries, and the specificity was good, a repeatability results showed that the three repeatability tests were consistent, with no false positives or false negatives; the sensitivity of this method for Gastrodia elata genomic DNA was 1 pg·μL^-1, which was higher than that of conventional PCR; the ERA technique can be used for the rapid identification of commercially available samples of Gastrodia elata and the results of the assay are the same as those of the PCR method. Conclusion: The established detection method is simple, rapid, with high specificity and sensitivity, and provides a new means for the authentication of the Gastrodiae Rhizoma.

The national drug sampling and inspection project is an important way of drug quality supervision in China which providing strong support for drug supervision and standard improvement. This article summarizes the national drug sampling and inspection project of Arnebiae Radix completed by Institute for Control of Chinese Traditional Medicine and Ethnic Medicine, National Institutes for Food and Drug Control in 2015 and 2022. The results illustrated that the qualification rate of Arnebiae Radix has increased from 43.9% in 2015 to 87.5%, and the qualification rate of Arnebiae Radix has significantly increased. The two nationwide inspections of Arnebiae Radix reflected the scarcity of Arnebiae Radix resources in Xinjiang and Inner Mongolia, resulting in a high market share of unqualified samples. The thin layer identification spots of the unqualified Arnebiae Radix sampled in 2015 were not consistent with the qualified samples. The thin layer identification of the unqualified samples sampled in 2022 was consistent with those of the qualified samples, but the depth of the spots were not consistent with those of the qualified samples, indicating that the current unqualified samples were adulterated samples, which posing greater challenges to the quality supervision of Arnebiae Radix. Through the exploratory research of twice National Drug Sampling and Inspection Project, it is preliminarily believed that it is of great significance to improve the standard test items of Arnebiae Radix, scientifically establish the limit value and strengthen the construction of the quality control system for the supervision.

Objective: To design and screen specific primers for efficient amplification and identification of Arnebiae Radix from market based on the concept of nested PCR. Methods: Nested primers was designed using the software of Primer Premier 5 based on the ITS sequence of Arnebia euchroma and the ITS2 sequence of non-pharmacopoeial Arnebiae Radix. The amplification efficiency of genomic DNA by ITS2 universal primers PCR and nested PCR was compared. The genomic DNA of Arnebiae Radix was amplified directly by nested primers and was detected by agarose gel electrophoresis. The specific primers designed for Arnebiae Radix based on the fragment length and variation sites’ coverage of the amplified product was evaluated. Results: A total of 11 primers were selected for synthesis after the primers were designed by Primer Premier 5 software. The amplification efficiency of nested PCR was superior to ITS2 universal primers PCR in genomic DNA of Arnebiae Radix. The results of nested primers directly amplified genomic DNA of Arnebiae Radix by agarose gel electrophoresis were better than those of ITS2 primers, and showed a single band. Four pairs of primers, AE-9S/AE-2A, AE-4S/AE-10A, AE-12S/10A, AE-29S/AE-29A, were determined to be suitable for the identification of Arnebiae Radix. Conclusion: On the basis of DNA barcode identification and nested PCR technology, 4 pairs of specific primers are identified which can be used to effectively distinguish Arnebia euchroma from the mainstreamed non-pharmacopoeial Arnebiae Radix in the medicinal materials market, providing reference for the subsequent research and development of identification methods for Arnebiae Radix and other traditional Chinese medicines.

Objective: To establish a method for simultaneous determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, cynarin, galuteolin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C in Shanjujiangya capsules by HPLC and analyze compositional change of Chrysanthemi Flos combined with law of quantity transfer. Methods: The analysis was performed on Waters Symmetry C₁₈ column (250 mm×4.6 mm,5 μm), with mobile phase composed of acetonitrile -0.1% phosphoric acid solution at a flow rate of 1.0 mL·min^-1 in gradient elution mode. The column temperature was 30 ℃ and the detection wavelength was 328 nm. The transfer rates of the above eight components were used as the indexes for quality evaluation to study the quantity value of transfer rule from the decoction piece to the extracting solution. Results: The results showed that the determination of eight components manifested a good linear relationship in the range of mass concentration (r>0.999 9), the average recoveries of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, cynarin, galuteolin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C were 98.3%-101.9%,with RSDs of 0.066%-0.64%. The contents of the above 8 components in 3 samples were 0.257-0.279 mg·g^-1, 0.629-0.650 mg·g^-1, 0.402-0.476 mg·g^-1, 0.454-0.539 mg·g^-1, 1.118-1.278 mg·g^-1, 0.653-0.740 mg·g^-1, 0.659-0.706, 1.138-1.167 mg·g^-1, respectively. Conclusion: The HPLC method established in this study is simple, repeatable and stable. The analysis of quantity transfer provides data support for the establishment of content methods and the formulation of limits. This study can provide basis for quality control method of Shanjujiangya capsules.

Objective: To establish an HPLC-MS/MS method for simultaneous determination of 11 components (harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone, harpagoside, vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ, and atractylenolide Ⅰ) in Gengnianning. Methods: HPLC assay was performed on C₁₈ column(100 mm×2.1 mm, 1.9 μm) with a mixture of methanol and 0.1% formic acid as the mobile phase in gradient elution at a flow rate of 0.3 mL·min^-1.The column temperature was 25 ℃ and the injection volume was 1 μL. Detection was carried out on a triple quadrupole mass spectrometer in positive ion mode (harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone and harpagoside) and negative ion mode(vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ, and atractylenolide Ⅰ) using an electrospray ion source(ESI). Multiple reaction monitoring(MRM)mode was employed. Results: The calibration curves were linear within the ranges of 1.485-29.71 μg·mL^-1, 1.620-32.40 μg·mL^-1, 7.801-156.0 μg·mL^-1, 0.518-10.35 μg·mL^-1, 0.167-3.333 μg·mL^-1, 0.359-7.179 μg·mL^-1, 1.455-29.10 μg·mL^-1, 1.520-30.40 μg·mL^-1, 0.160-3.205 μg·mL^-1, 0.143-2.864 μg·mL^-1 and 0.157-3.136 μg·mL^-1 for harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone, harpagoside, vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ, and atractylenolide Ⅰ, respectively. All 11 components showed good linearity (r≥0.998 0). The average recoveries(n=6)were in the range of 95.9%-102.6% with RSDs within 0.90%-3.0%. The contents of harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone, harpagoside, vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ and atractylenolide Ⅰ in 10 tested samples from 5 manufactures were 14.8-104.5, 37.6-288.5, 335.8-1 332.8, 6.2-10.1, 6.6-61.8, 13.7-75.1, 57.4-132.8, 16.9-70.6, 11.8-33.9, 3.4-15.4 and 6.5-12.9 μg·g^-1. Conclusion: The developed method is accurate and sensitive. It can be used in quality control of Gengnianning.

Objective: To establish a ultra-high performance liquid chromatography-mass spectrum in series method for the quantification of 7-hydroxycoumarin, fraxetin, isofraxidin and scopoletin in the aqueous extract of Acanthopanacis Senticosi Radix et Rhizoma Seu Caulis in rat plasma, and to evaluate the pharmacokinetic behavior and bioavailability in rats. Methods: The chromatography was performed on Thermo Scientific Hypersil GOLD aQ (100 mm×2.1 mm, 1.9 μm)column with 0.1% aqueous formic acid -0.1% acetonitrile formic acid solution (82∶18) as the mobile phase at the flow rate of 0.30 mL·min^-1, and column temperature of 40 ℃. Mass spectrometry positive ion mode scan was used, and sample size was 5 μL. Healthy SD rats were selected for a single gavage of 10 mL·kg^-1 of Acanthopanacis Senticosi Radix et Rhizoma Seu Caulis water extract (equivalent to the dose of 1 g·kg^-1 of the original drug). Plasma concentration of the substances was determined at different time intervals after administration, and the pharmacokinetic parameters were calculated using DAS software by non-atrioventricular model fitting. Results: The methodological results showed that 7-hydroxycoumarin (r=0.999 4), fraxetin (r=0.998 9), isofraxidin (r=0.999 3) and scopoletin (r=0.998 4) had good linearity in the range of 0.05-55 μg·mL^-1, and RSDs of precision of the four substances were all less than 15%. The recovery was 85%-115%. The absolute bioavailability ranged from 59% to 78%, and the relative bioavailability ranged from 80% to 87%. The extraction recovery, matrix effect and stability met the relevant requirements. Conclusions: After a single gavage of the aqueous extract of Acanthopanacis Senticosi Radix et Rhizoma Seu Caulis in rats, the four substances to be tested are absorbed and eliminated by rats. The method validation results are in line with the guiding principles of biological sample analysis methods, and can be applied to evaluate the pharmacokinetic behavior and bioavailability of the aqueous extract of Acanthopanacis Senticosi Radix et Rhizoma Seu Caulis in rats.

Objective: To establish a three classification model for cultivated, semi-wild, and wild Astragali Radix characterized by flavonoids, and explore and evaluate the application of techniques of automated machine learning and data augmentation in the field of drug analysis. Methods: Firstly, correlation analysis and principal component analysis were conducted on the flavonoid content data of Astragali Radix, and models of decision tree and logistic regression were established to analyze the importance of flavonoid components based on the models. Then, using the AutoGluon framework with 5 as num_bag_folds, 2 sets of 30 models respectively through 64 batches of real data and 600 batches of virtual data generated based on real data with the TVAE table data generation algorithm for training were obtained, and these models were evaluated by accuracy. Results: The analysis of machine learning models, indicated that formononetin, campanulin and onospin played the important roles in the quality control of Astragali Radix, especially for the source grade control. The accuracy of model prediction showed that the models based on Neural Net and tree-model always had the best classification effect for Astragali Radix. The virtual data generated by data augmentation technique is basically consistent with the actual data in terms of the accuracy trend of the model training process. Conclusion: Related techniques of machine learning have good application value in the classification of Astragali Radix characterized by flavonoids.

Objective: To base on the method of HPLC fingerprint, multi-component quantification and chemical pattern recognition, to evaluate the quality of leaves of Cunninghamia lanceolata from different producing areas and to provide basis for further development and utilization. Methods: High-performance liquid chromatography (HPLC) was used to determine the contents of amentoflavone, bilobetin, hinokiflavone, ginkgetin, isoginkgetin and sciadopitysin in the Cunninghamia lanceolata. Fingerprints of 10 batches of Cunninghamia lanceolata from different habitats were established. Based on the common peak area of the fingerprint, the overall quality of Cunninghamia lanceolata was evaluated by principal component analysis(PCA), orthogonal partial least squares discriminant analysis(OPLS-DA), statistical analysis, and pattern recognition chemometrics methods. Results: A total of 14 common peaks in 10 batches of leaves of Cunninghamia lanceolata, and the similarity ranged from 0.955 to 1.000 with good consistency. The mass fractions of six biflavones in the sample were 2.42-5.24 mg·g^-1, 0.10-0.24 mg·g^-1, 1.55-3.67 mg·g^-1, 0.21-0.89 mg·g^-1, 0.10-0.24 mg·g^-1 and 0.51-2.39 mg·g^-1, respectively, including amentoflavone, bilobetin, hinokiflavone, ginkgetin, isoginkgetin and sciadopitysin. According PCA, the difference in the quality of Cunninghamia lanceolata from different habitats was further evaluated. Ten batches of medicinal materials were divided into three major categories, and four main factors affecting the classification of Cunninghamia lanceolata were found. Finally, OPLS-DA screened the excellent spectral peaks 6, 12, 7 (7-demethylated ginkgo biloba biflavone), 13 (kumatsu biflavone), 5 (Amentotaxus argotaenia biflavone), 9 (chamaecypress biflavone) and 2, etc, seven differential markers can be used to distinguish different batches of Cunninghamia lanceolata. Conclusion: The established method is simple to stable and reliable. Combined with chemical pattern recognition, it can be used for the quality evaluation of Cunninghamia lanceolata.