Latest ArticlesStereoisomers are a common form in natural products，and the pharmacological activities of isomers with different configurations are various. Efficient separation and analysis of stereoisomers is one of the urgent problems for the study of pharmacodynamic substances of natural products. In order to comprehensively and systematically review the separation and analysis methods of stereoisomers，this review used online databases PubMed，Web of Science，Google Scholar，ACS，CNKI，and others to comprehensive search for the separation and analysis of the isomeric，and systematically summaries the advantages，disadvantages and scope of application. The results showed that the separation methods of stereoisomers mainly included high performance liquid chromatography，two-dimensional liquid chromatography，supercritical fluid chromatography，gas chromatography and capillary electrophoresis. In addition，non-chromatographic methods such as preferential crystallization，membrane separation and kinetic splitting can also be used for stereoisomers splitting. Commonly used stereoisomer resolution methods include conventional mass spectrometry，ion mobility mass spectrometry，nuclear magnetic resonance techniques，single-crystal X-ray diffraction and spectroscopy. The study of common stereoisomer separation and analysis methods is summarized with a view to laying a foundation for the research and application of stereoisomers in natural products.
To establish the determination methods for chromatographic fingerprint and multiple constituents，and then evaluate the quality grade of 103 batches of Banlangen granules that were from 55 manufacturers based on the reference drug.
Ultra-high-performance liquid chromatography (UPLC) analysis was used to establish the fingerprint of Banlangen granules and quantitate the contents of 7 components-uridine，adenine，vernine，(R，S)-goitrin，adenosine，syringin and clemastanin B. The test was performed on the Waters ACQUITY UPLC HSS T3 (100 mm×2.1 mm，1.8 μm) column with gradient elution using methanol-water at a flow rate of 0.2 mL·min-1. The injection volume was 2 μL，and the column temperature was constant at 30 ℃. Then，the results were detected using adopting a multi-wavelength detection mode. Next，the characteristic peaks and their belonging were clarified by comparing them with the reference drug. The sample fingerprint similarity and determination of contents were finally calculated，and the quality grades of Banlangen granules in each item were evaluated by using the reference drug.
The fingerprint was established，and 10 main characteristic peaks were identified. The similarities of the sample’s fingerprints ranged from 0.541 to 0.993，of which 101 batches were over 0.75 that were reached the second level，and 81 batches were over 0.90 that were reached the first level. The linear ranges of 7 components had good linear relationships within their respective ranges. The average recoveries (n=9) were 97.0%-104.7% with RSDs all below 3%. The precision，stability，and repeatability of methods were all good with RSDs all below 3%. Unified the sample specification to 5 g per bag，the content range of Banlangen granules samples was between 0.189-10.347 mg per bag. As a result，the content of 75 batches of samples reached the second level，and the content of 59 batches of samples reached the first level.
The established methods are simple，accurate and rapid，which can be used for the quality control and grade evaluation of Banlangen granules，as well as the research paradigm of other traditional Chinese medicine preparations.
To establish an ICP-MS method for the detection of elemental impurities in lanthanum oxide and to explore the development of the ICP-MS methodology for high-matrix samples quantitative analysis.
25 mg samples were precisely weighed and placed in a 25 mL flask. 1 mL of nitric acid and 0.25 mL of hydrochloric acid solution were added，shaken，and dissolved for approximately 1 h，then fixed with ultrapure water. Quantitative samples determination were performed using Agilent 7900 ICP-MS in He mode with the standard curve method corrected by internal standard. The atomizing gas flow rate was set at 1.05 L·min-1，the atomizing chamber temperature was maintained at 2 ℃，the sample aspiration rate was 0.1 r·s-1，the RF power was 1 550 W，and the sampling depth was 10 mm. The helium flow rate was 5 mL·min-1，and the energy discrimination was 5.0 V. Based on ICH Q2 (R2) and USP 2023 <233>，a quantitative method for 25 elemental impurities was established by utilizing the recovery rate of the method. The isotope ratio of each element in the sample was investigated，the mass number of the element to be measured was selected，and the method was verified.
Based on the specific outcomes of Se and Ce，82 and 142 were chosen as the detection mass numbers to evade interference. The results demonstrated that the linear relationships of the 24 elemental impurities were excellent，the recoveries ranged from 70% to 150%，the repeatability RSDs were less than 20%，and the method satisfied the quality control requirements. Impurities such as Pb (0.152-0.201 ng·g-1)，Cd (0.007-0.010 ng·g-1)，Hg (0.156-0.250 ng·g-1) were detected in numerous batches of samples，and the contents of each element were lower than the proposed standard.
The method is specific，accurate，simple，and feasible，and can furnish technical support for the elemental impurity control of lanthanum oxide. For complex and high-matrix samples，the recovery rate of standard addition is inadequate to characterize the specificity and accuracy of the method，and the isotope ratio can be utilized as a supplement to the recovery rate of standard addition.
To develop a method for the determination of meropenem and vaborbactam for injection by HPLC.
The chromatography was performed on Shimadzu InertSustain C18 (150 mm×4.6 mm，5 μm) column with acetonitrile-0.02 mol·L-1 sodium dihydrogen phosphate solution (adjust pH to 2.8 with phosphoric acid)(10：90) as a mobile phase，the flow rate was 1.0 mL·min-1，the detection wavelength was 210 nm，and the sample volume was 10 μL. A method for the determination of meropenem and vaborbactam for injection was established under the chromatographic conditions.
In this method，the linearities of meropenem and vaborbactam were good，and the linearity ranges were 21.40-214.0 μg·mL-1 and 19.83-198.3 μg·mL-1. Meropenem and vaborbactam had good precision with RSD of 0.31% and 0.16%，respectively. Within 24 h，meropenem and vaborbactam had good stabilities at 4 ℃，RSDs were 0.31% and 0.16%，respectively. The average recoveries of meropenem and vaborbactam were 101.0% (n=9) and 98.4% (n=9) (RSD<2.0%). The contents of meropenem and vaborbactam were 405.8 mg·g-1 and 399.1 mg·g-1，respectively.
The method is accurate，simple and rapid，and can be used for the determination of meropenem and vaborbactam for injection.
To establish an HPLC method for the ratio of the enantiomer in fluralaner.
The chromatographic separation was performed on chiral chromatography column CHIRALPAK AD-H (250 mm×4.6 mm，5 μm). The mobile phase consisted of n-hexane-anhydrous ethanol (60：40) and the flow rate was 1.0 mL·min-1. The detection wavelength was 265 nm and the column temperature was room temperature.
Under this chromatographic condition，three batches of samples were determined，the proportions of R-fluralaner and S-fluralaner were 1：1. The calibration curves of R-fluralaner and S-fluralaner good linearities at the range of 80.288-187.338 μg·mL-1(r=0.999 7) and 81.902-191.104 μg·mL-1 (r=0.999 9)，respectively. The average recoveries were 100.6% and 100.8%.
The method is accurate，reproducible and can be used for the ratio of enantiomers in fluralaner racemate.
Objective: To establish the HPLC fingerprints for Zilian Shengji gel and to determine the contents of gallic acid, tableberberine, coptisine, palmatine, berberine and shikonin. Methods: The establishment of fingerprints was performed on a 25 ℃ thermostatic Agilent C18-WR（250 mm×4.6 mm, 5 μm）column with the mobile phase comprising of acetonitrile -0.15% phosphoric acid water at the flowing rate of 0.8mL·min-1 in a gradient elution manner, and the detection wavelength were set at 270 nm, 345 nm and 516 nm. Cluster analysis, principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were adopted in chemical pattern recognition. The six components identified were quantitatively determined. Results: The HPLC fingerprint of Zilian Shengji gel was constructed with twenty-six common chromatographic peaks, eleven batches of samples with the similarities of 0.828-0.997, six chromatographic peaks of gallic acid, tableberberine, coptisine, palmatine, berberine and shikonin were identified by reference substance comparison. Six constituents showed good linear relationships within their own ranges (r≥0.999 5) whose average recoveries were 98.7%-99.3%, with the RSDs of 1.3%-1.8%. The content ranges of the above mentioned six components in eleven batches of samples were 827.74-1 513.50 μg·g-1, 137.52-296.05 μg·g-1, 381.83-884.73 μg·g-1, 2 023.81-4 051.66 μg·g-1, 524.15-986.13 μg·g-1 and 47.65-549.46 μg·g-1. Conclusion: This simple, stable, reliable and reproducible method can be used for the quality control of Zilian Shengji gel.
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 establish the infrared spectrum identification model to distinguish Mingui from Angelica sinensis from other habitats. Methods: A total of 1 540 batches of A. sinensis were collected from Gansu Province through visits and field investigation, and the mid-infrared spectrum of each sample was collected by Fourier transform infrared spectroscopy. The average spectra of 423 batches of Mingui were calculated as their fingerprint. The identification threshold of Mingui was determined as 0.989 6 (μ-3σ) by mid-infrared spectroscopy combined with the “3σ” criterion. The identification model of Mingui was established. Three batches of Mingui and 97 batches of A. sinensis from other habitats were randomly selected to verify the identification model. Results: The characteristic peaks of samples were observed at 3 560, 3 389, 3 342, 2 931, 1 743, 1 650, 1 459, 1 409, 1 385, 1 374, 1 323, 1 279, 1 239, 1 128, 1 115, 1 104, 1 068, 1 053, 1 013, 1 004, 991 and 920 cm-1, etc. The mid-infrared spectra of 423 batches of Mingui were highly similar, and their peak shape, peak position and peak intensity were similar. The correlation coefficients between Mingui and the fingerprint ranged from 0.992 0 to 0.998 0. The accuracy of identification model test was 97%. Conclusion: The identification model of Mingui could be used to distinguish Mingui from other A. sinensis from other habitats.
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 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.
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