HPLC analysis of 16 compounds from Artemisia ordosica / 中草药·英文版

OBJECTIVE@#To establish a high-performance liquid chromatographic method (HPLC) for the simultaneous determination of 16 compounds from Artemisia ordosica.@*METHODS@#HPLC was used to analyze 16 quality indicators of A. ordosica. The HPLC conditions were as follows: Agilent Eclipse Plus C18 column (250 mm × 4.6 mm, 5 μm) with acetonitrile (A)-water (B) as mobile phase, gradient elution: 0-10 min, 75%-65% B; 10-30 min, 65%-35% B; and finally 30-40 min, 35%-15% B. The flow rate was 1.0 mL/min, the column temperature was 40 °C, the injection volume was 10 μL, and monitored by absorbance at 285 nm for compounds 1- 10, 12 and 225 nm for compounds 11, 13- 16.@*RESULTS@#Under the selected experimental chromatographic conditions, compounds 1- 16 showed good linearity (r > 0.9993) in a wide concentration range. Their average recoveries were 99.50%, 95.38%, 97.75%, 96.00%, 98.20%, 97.50%, 95.50%, 99.33%, 96.75%, 96.50%, 98.50%, 97.83%, 99.20%, 95.33%, 97.33% and 96.30%, respectively, and the RSD were 1.99%, 1.81%, 1.63%, 1.98%, 1.67%, 1.92%, 1.74%, 1.67%, 1.90%, 1.72%, 1.88%, 1.83%, 1.79%, 1.76%, 1.81% and 1.96%, respectively.@*CONCLUSION@#Based on the results of the HPLC analysis, it was concluded that p-hydroxycinnamic acid ( 1), O-hydroxycinnamic acid ( 2), coniferyl alcohol ( 5), 5,4'-dihydroxy-7,3'-dimethoxyflavanone ( 8), 5,4'-dihydroxy-7-methoxyflavanone ( 9), 5-hydroxy-7,4'-dimethoxyflavanone ( 12), dehydrofalcarindiol ( 13), arteordoyn A ( 14), dehydrofalcarinol ( 15) and capillarin ( 16) are best suited for the role of quality indicators of A. ordosica grown in different ecological environments.

Nitrogen-doped carbon@TiO2 double-shelled hollow spheres as an electrochemical sensor for simultaneous determination of dopamine and paracetamol in human serum and saliva / 药物分析学报

As the most commonly used antipyretic and analgesic drug,paracetamol(PA)coexists with neuro-transmitter dopamine(DA)in real biological samples.Their simultaneous determination is extremely important for human health,but they also interfere with each other.In order to improve the conductivity,adsorption affinity,sensitivity,and selectivity of TiO2-based electrochemical sensor,N-doped carbon@-TiO2 double-shelled hollow sphere(H-C/N@TiO2)is designed and synthesized by simple alcoholic and hydrothermal method,using polystyrene sphere(PS)as a template.Meanwhile,TiO2 hollow spheres(H-TiO2)or N-doped carbon hollow spheres(H-C/N)are also prepared by the same method.H-C/N@TiO2 has good conductivity,charge separation,and the highly enhanced and stable current responses for the detection of PA and DA.The detection limit and linear range are 50.0 nmol/L and 0.3-50 μmol/L for PA,40.0 nmol/L and 0.3-50 μmol/L for DA,respectively,which are better than those of carbon-based sen-sors.Moreover,this electrochemical sensor,with high selectivity,strong anti-interference,high reli-ability,and long time durability,can be used for the simultaneous detection of PA and DA in human blood serum and saliva.The high electrochemical performance of H-C/N@TiO2 is attributed to the multi-functional combination of different layers,because of good conductivity,absorption and electrons transfer ability from in-situ N-doped carbon and electrocatalytic activity from TiO2.

Stability-indicating HPLC-DAD method for the simultaneous determination of fluoroquinolone in combination with a non-steroidal anti-inflammatory drug in pharmaceutical formulation

We developed and validated a stability-indicating assay method for the simultaneous determination of enrofloxacin and piroxicam in combination and in the presence of degradation products. Reverse-phase high-performance liquid chromatography analyses were carried out on a Vertisep C18 column and acetonitrile-water (48:52 v/v, pH 3.0) mobile phase with a 1.00 mL min−1 flow rate. The efficient chromatographic separation of these drugs and their forced degradation products was achieved in less than 5min with a peak purity match factor higher than 950. The method used showed linearity in the concentration ranges of 0.25 to 16.0 µg mL−1 for enrofloxacin (r = 0.9997) and 0.125 to 8.0 µg mL−1 for piroxicam (r = 0.9999) as well as precision (relative standard deviation lower than 2%), accuracy (mean recovery 100 ± 2%), and robustness, according to ICH (International Conference on Harmonization) and AOAC (Association of Official Analytical Chemists) guidelines. This method can simultaneously determine the combination of these drugs in a veterinary formulation and separate the drug peaks from their forced degradation products. Additionally, its optimized chromatographic conditions can contribute to the quality control of this formulation in pharmaceutical manufacturing plants and minimize waste from the organic solvent.

Quality evaluation of Magnoliae Officinalis Cortex based on combinative method of fingerprint,quantitative analysis of multicomponents and chemometrics / 中国中药杂志

This study aims to establish a combinative method based on fingerprint,assay of multi-component and chemometrics for quality evaluation of Magnoliae Officinalis Cortex. Twenty batches of samples were determined by UPLC and a common mode of fingerprint was established. The similarities between fingerprints of 20 batches of samples were over 0. 90 and the common mode were evaluated. Eight components were identified as syringing, magnocurarine, magnoflorine, magnoloside B, magnoloside A, honokiol,magnolol,and piperitylmagnolol by comparison with reference substances and their content in samples were simultaneously determined.Based on the results,the fingerprint had good consistency between the same origin and minor diversity between the different sources.Piperitylmagnolol and peak 13 could be used as a distinction with the different sources. According to content of 8 components,Fisher discriminant analysis model was established and different source sample was classified pursuant to the discriminant fraction. It is indicated that simultaneous quantification of multi components coupled with chemometrics analysis could be a well-acceptable strategy to identify and evaluate the quality of Magnoliae Officinalis Cortex.

Highly sensitive simultaneous electrochemical determination of myricetin and rutin via solid phase extraction on a ternary Pt@r-GO@MWCNTs nanocomposite / 药物分析学报

The simultaneous electrochemical determination of myricetin and rutin remains a challenge due to their indistinguishable potentials. To solve this problem, we constructed a ternary platinum nanoparticle, reduced graphene oxide, multi-walled carbon nanotubes (Pt@r-GO@MWCNTs) nanocomposite via a facile one-pot synthetic method. Under the optimized conditions, the ternary Pt@r-GO@MWCNTs nanocomposite exhibited good electrocatalytic activity toward myricetin and rutin via solid phase extraction and excellent performance for the simultaneous determination of myricetin and rutin. The oxidation peak current of myricetin was proportional to its concentrations in the range of 0.05-50μM with a detection limit of 0.01μM (S/N = 3). The linear range for rutin was 0.05-50μM with a detection limit of 0.005μM (S/N = 3). The ternary nanocomposite sensor also exhibited good reproducibility and stability, and was successfully used for the simultaneous determination of myricetin and rutin in real orange juice samples with recoveries ranging between 100.57％and 108.46％.

Simultaneous Determination of Twelve Kinds of Residual Solvents in A New Drug CBT108 By Headspace Gas Chromatography / 分析化学

A simple and sensitive method for simultaneous determination of 12 kinds of residual solvents in a new drug CBT108 was established and validated by headspace gas chromatographic technology. The rationality,accuracy and feasibility of the analytical method were verified. Under the optimized conditions, simultaneous separation and determination of 12 kinds of residual solvents, including methanol, ethanol, ether, acetone, acetonitrile, dichloromethane, n-hexane, ethyl acetate, tetrahydrofuran, heptane, toluene and carbon tetrachloride was carried out by using a DB624 capillary column(30 m×0.53 mm×3.0 μm) for separation, a flame ionization detector for detection and internal standard method for quantitation. Good linearity was obtained for 12 solvents with the correlation coefficients(R2) of more than 0.997. The limits of quantitation and detection were defined at S/N=3 and S/N=10,respectively. LOQ and LOD for 12 solvents were given as 0.024 μg/mL and 0.0072 μg/mL for methanol,0.1 μg/mL and 0.012 μg/mL for ethanol, 0.01 μg/mL and 0.005 μg/mL for ether, 0.1 μg/mL and 0.008 μg/mL for acetone, 1.025 μg/mL and 0.0615 μg/mL for acetonitrile, 0. 09 μg/mL and 0. 06 μg/mL for dichloromethane, 0. 09 μg/mL and 0.06 μg/mL for n-hexane, 0. 25 μg/mL and 0. 008 μg/mL for ethyl acetate, 0. 108 μg/mL and 0.014 μg/mL for tetrahydrofuran,0.16 μg/mL and 0.0004 μg/mL for carbon tetrachloride,0.0075 μg/mL and 0.005 μg/mL for heptane, and 0.0445 μg/mL and 0.0014 μg/mL for toluene. The adding standards recoveries for 12 residual solvents at three spiked levels were in the range of 90.96%-108.67%,with relative standard deviations of 0.1%-5.7%. This simple,high accuracy and good repeatability method is feasible for rapidly determination of 12 residual solvents in drug candidate CBT108. Meanwhile, this simple method provides a consulted value for detection of residual solvents in other medicines.

Comparative chemical characters of Sparganii Rhizoma from different regions in China / 中草药·英文版

Objective: To compare the chemical characters of Sparganii Rhizoma from different areas via chromatographic analysis and to establish a sensitive LC/MS method for quality assessment of Sparganii Rhizoma. Methods: Under the optimised HPLC-PDA chromatographic conditions, twenty batches of Sparganii Rhizoma were analyzed by HPLC fingerprints. Principal component analysis (PCA), orthogonal projections to latent structures discriminant analysis (OPLS-DA) and hierarchical cluster analysis (HCA) were performed based on all peak areas of Sparganii Rhizoma fingerprints. Meanwhile, part of common peaks were subsequently quantified by UFLC-QTRAP-MS. Results: The similarity values of HPLC fingerprints fluctuated in a wide range of 0.511–0.973, which showed variable differences of chemical characters among Sparganii Rhizoma from twenty habitats. PCA, OPLS-DA and HCA indicated that samples could be divided into five groups with different chemical characters, which generally corresponded with their geographical distributions. A total of 31 peaks in HPLC fingerprints were marked, and eight of them were identified and quantified. The quantitative result was generally in agreement with the classifications based on HPLC fingerprints, which indicated that Sparganii Rhizoma samples from eastern China mostly contained more contents including phenolic acids and flavonoids. Conclusion: This study not only proved that there were relationships between geographic distributions and internal chemical compositions of plants, which could provide evidence to the traditional Chinese medicine concept “geo-authentic” but also supplied a sensitive and rapid simultaneous quantitive method for the quality estimation of Sparganii Rhizoma.

Ion Chromatography Method of Four Carboxylic Acids in Workplace Air / 中华劳动卫生职业病杂志

Objective@#To assess psychological acceptance and occupational stress of medical staff, analyze the relationship among personality, psychological acceptance and occupational stress and discuss the direct or indirect effects of personality to occupational stress.@*Methods@#The gaseous four kinds of carboxylic acids in the workplace air were simultaneously collected by silica gel tube, and then desorbed by deionized water and eluted by ion chromatograph. The the content was detected by conductivity detector.@*Results@#The linear relationship was good in the concentration range of about 0~140 mg/L. The correlation coefficient r>0.999, and the maximum detection limit was 4.5 μg/mL. The sampling efficiency of the four carboxylic acids ranges from 96.10%~100.27%. Through the sample added recovery experiments, the low and high content of the silicone tube samples were detected; and the range of desorption efficiency was 82.18%~100.12%; the range of precision was 0.70%~3.71%.@*Conclusion@#This method adopts deionized water to desorb samples, and the application of ion chromatography detection have reached the requirements of《Guidelines for the establishment of occupational health standards, Part 4: Determination of chemical substances in workplace air》, which can be used in four kinds of workplace air detection of carboxylic acid compounds.

Content Determination and Cluster Analysis of 3 Lignans from Cortex Magnoliae Officinalis in Different Regions / 世界科学技术-中医药现代化

Objective: To establish the method of simultaneous determination of contents of honokiol, magnolol and piperitylmagnolol in Cortex Magnoliae Officinalis from different regions, which could provide evidence for the quality control evaluation of Cortex Magnoliae Officinalis. Methods: The quantitative analysis was performed with a column of Waters Acquity UPLC BEH-Cl8 (2.1 mm×50 mm, 1.7 μm) by UPLC-PDA, and eluted with a mobile phase of 0.2％formic acid solution (A)-methanol (B) in a gradient mode (0-2 min, 65％-65％ B; 2-3 min, 65％-75％ B; 3-5 min, 75％-84％ B; 5-9 min, 84％-90％ B) under a flow rate of 0.5 mL·min-1. The column temperature was set at 35℃, and the detection wavelength was 294 nm and the injection volume was 1 μL. Results: The 3 lignan components were completely separated and could be separated well from other components. The honokiol, magnolol and piperitylmagnolol showed a good linear relationship with chromatographic peak area in range of 20.3-406.0, 15.2-304.0, 5.6-112.0 ng, respectively.The average recoveries were 91.75％, 93.86％, 95.15％ with the RSDs were 1.75％, 1.88％, 1.91％ (n = 9), respectively.Conclusion: Contents of the 3 constituents from different place were significantly different and this method is simple and effective, which can be used to quality control of Cortex Magnoliae Officinalis.

Simultaneous determination of twelve constituents in nüjin pills by hplc combined with wavelength switching / 中国药学杂志

OBJECTIVE: To establish an HPLC method for simultaneous determination of 12 constituents in Nüjin pills. METHODS: The analysis was carried out on Agilent Eclipse plus C18 column(4.6 mm×250 mm,5 μm), using a mobile phase of acetonitrile (A)-0.2%formic acid solution (B) by a gradient elution program (0-25 min, 10%-35% A; 25-37 min, 35%-59% A; 37-40 min, 59%-75% A) at a flow rate of 1 mL•min-1 at 30 ℃. The detection wavelength was set at 230 nm in the first 20 min, and then changed to 270 nm between 20 and 40 min. RESULTS: The linear ranges of paeoniflorin, liquiritin, ferulicacid, narirutin, aurantiamarin, baicalin, wogonoside, cinnam aldehyde, baicalein, paeonol, ammonium glycyrrhetate and wogonin fell within the ranges of 0.011-1.062 μg, 0.002-0.237 μg, 0.003-0.259 μg, 0.006-0.641 μg, 0.029-2.925 μg, 0.033-3.339 μg, 0.008-0.834 μg, 0.003-0.258 μg, 0.008-0.772 μg, 0.008-0.824 μg, 0.007-0.724 μg and 0.005-0.539 μg, respectively. The recoveries were 99.3%, 98.4%, 99.1%, 99.4%, 99.5%, 99.6%, 99.4%, 99.2%, 99.0%, 99.6%, 99.1%, and 99.4%, respectively. The relative standard deviations were 0.35%, 1.02%, 0.33%, 0.38%, 0.18%, 0.27%, 0.40%, 0.74%, 0.53%, 0.16%, 0.08% and 0.21%, respectively (n=6). CONCLUSION: This method is simple, accurate and convenient for the quality control of Nüjin pills.

Simultaneous determination of amino acids and nucleosides of three species in Termitomyces by UFLC-QTRAP-MS-MS / 中国中药杂志

A comprehensive analytical method based on UFLC-QTRAP-MS-MS was developed for the simultaneous determination of 15 kinds of amino acids and 12 kinds of nucleosides of three species in Termitomyces. The separation was carried out on a Waters XBridge Amide column (2.1 mm×100 mm,3.5 μm) with gradient elution of mobile phase of 0.2% formic acid in water-0.2% formic acid in acetonitrile at a flow rate of 0.6 mL•min⁻¹, and column temperature was maintained at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. The principal component analysis(PCA) was made to standardized treatment for the comprehensive evaluation of different species in Termitomyces. The 15 kinds of amino acids and 12 kinds of nucleosides multiple constituents showed good linearity (r>0.997 3) in the range of the tested concentration.The average recoveries ranged from 95.14% to 105.0%,and the relative standard deviations were less than 5.0%. The comprehensive evaluation index obtained with PCA showed that the Termitomyces albuminosus was significantly higher than others in amino acids and in nucleosides, of which the T. aurantiacus was the best. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances,which provided a new basis for the comprehensive assessment and overall control of the quality of Termitomyces fungi.

Simultaneous determination of five furostanol saponins in rhizome and fibrous root of Anemarrhena asphodeloides Bge. by HPLC-CAD / 国际药学研究杂志

Objective To develop a method for the determination of five furostanol saponins(timosaponin N,timosaponin L, timosaponin BⅡ,25R-timosaponin BⅡ,and 25S-officinalisnin-Ⅰ)in rhizome and fibrous root of Anemarrhena asphodeloides Bge. by HPLC with the charged aerosol detector(CAD). Methods The analysis was performed on TechMate C18-ST-II(250 mm×4.6 mm,5μm)with acetonitrile:water(22:78,V/V),the flow rate of 1.0 ml/min and column temperature at 30℃. The Corona parameters were as follows:sampling rate 10 Hz,filter 5 s,and the nebulizer temperature 55℃. Results The approach showed good linearity for five saponins. The correlation coefficients(r2)for calibration curves varied from 0.9992 to 0.9998. The limits of detection(LOD)were 0.28,0.92,0.92,0.92 and 0.92 ng for five steroidal saponins,respectively. The limits of quantitation(LOQ)were found to be 0.92, 2.77,2.77,2.77 and 2.76 ng,respectively. RSD calculated from peak area of precision,repeatability and stability in 48 h were all less than 3.0%. The average recoveries of timosaponin N,timosaponin L,timosaponin BⅡ,25R-timosaponinBⅡ,and 25S-officinalis-nin-Ⅰwere 98.17%,101.37%,98.53%,97.63%,and 98.17%,respectively. Conclusion The developed method is accurate,reli-able,which could be applied to the quality control of multiple components in A. asphodeloides Bge.

Square wave voltammetric quantification of folic acid, uric acid and ascorbic acid in biological matrix / 药物分析学报

Nowadays, modified electrodes with metal nanoparticles have appeared as an alternative for the electroanalysis of various compounds. In this study, gold nanoparticles (GNPs) were chosen as interesting metal nanoparticles for modifying carbon paste electrode (CPE). GNPs and the gold nanoparticles-modified carbon paste electrode (GNPs/CPE) were characterized by UV–Vis spectroscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). GNPs/CPE as a simple and sensitive electrode was used to study three important biological molecules:folic acid (FA), uric acid (UA) and ascorbic acid (AA). Square wave voltammetry (SWV) was used as an accurate technique for quantitative measurements. A good linear relation was observed between anodic peak current (ipa) and FA (5.2 × 10?6–2.5 × 10?5 M), UA (1.2 × 10?6–2.1 × 10?5 M) and AA (1.2 × 10?6–2.5 × 10?5 M) concentrations in simultaneous determination of these molecules.

Simultaneous Determination of Saponins in Dripping Pills Made from Astragali Radix and Panax notoginseng by UPLC-ELSD / 中草药·英文版

Objective To develop a simple and fast method for removing polyethylene glycol (PEG) and simultaneous determination of fives saponins, i.e. astragaloside IV, notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1, and ginsenoside Rd in dripping pills made from Astragali Radix and Panax notoginseng. Methods The extraction method was based on a liquid-liquid extraction using water-saturated n-butanol and the quantitative determination was based on ultra-performance liquid chromatography coupled with evaporative light scattering detection (UPLC-ELSD). The chromatographic analysis was performed on an Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 µm) with a gradient elution of acetonitrile-0.1% formic acid aqueous solution within a runtime of 15 min. Results Compared to different methods, the proposed method could remove the interference of PEG in formulation. And the calibration curves showed good linearity during the test ranges. The method was validated for limits of detection and quantification, precision, and reproducibility. The recoveries were within the range of 96.87% – 99.97%. In addition, the verified method was firstly applied to determination of the five active ingredients in Qishen Yiqi Dripping Pills (QYDP) simultaneously. Conclusion The contents of five active ingredients are stable and homogeneous in QYDP, which indicates that the method could be readily utilized as a quality evaluation method for this traditional Chinese medicine dripping pills made from Astragali Radix and Panax notoginseng.

Simultaneous colorimetric determination of morphine and ibuprofen based on the aggregation of gold nanoparticles using partial least square / 药物分析学报

In this work a new method is presented for simultaneous colorimetric determination of morphine(MOR)and ibuprofen(IBU)based on the aggregation of citrate-capped gold nanoparticles(AuNPs).Citrate-capped AuNPs were aggregated in the presence of MOR and IBU. The difference in kinetics of AuNPs aggregation in the presence of MOR/IBU was used for simultaneous analysis of MOR and IBU. The formation and size of synthesized AuNPs and the aggregated forms were monitored by infra-red(IR)spectroscopy and transmission electron microscopy(TEM),respectively.By adding MOR or IBU the absorbance was decreased at 520 nm and increased at 620 nm.The difference in kinetic profiles of aggregation was applied for simultaneous analysis of MOR and IBU using partial least square(PLS)regression as an efficient multivariate calibration method.The number of PLS latent variables was optimized by leave-one-out cross-validation method using predicted residual error sum of square. The proposed model exhibited a high capability in simultaneous prediction of MOR and IBU concentrations in real samples. The results showed linear ranges of 1.33–33.29μg/mL (R2=0.9904) and 0.28–6.9μg/mL (R2=0.9902) for MOR and IBU respectively with low detection limits of 0.15 and 0.03μg/mL(S/N=5).

Simultaneous determination of xylitol and L-xylulose in fermentation broth with high performance liquid chromatography / 中国生化药物杂志

Objective A high performance liquid chromatographic (HPLC) method was established for the simultaneous determination of xylitol and L-xylulose in fermentation broth.Methods The chromatographic conditions were as follows:C18 column (250 mm ×4.6 mm) with the temperature 35℃, acetonitrile-water (85∶15,v/v)as mobile phase with the flow rate of 0.8 mL/min.Xylitol was detected by refractive index (RI) detector at 33℃and L-xylulose was determined by ultraviolet ( UV) detector at 210 nm at room temperature.Results This method showed good linearity over the range from 0.50~30.00 g/L with a correlation coefficient of 0.9995 for xylitol and 0.30~30.00 g/L with a correlation coefficient of 0.9986 for L-xylulose. Moreover, the limit of quantification (LOQ) for xylitol and L-xylulose were 0.58 and 0.40,respectively.The limit of determination (LOD) for xylitol and L-xylulose were 0.18 and 0.15,respectively.The relative standard deviations (RSDs) of intraday and interday for xylitol were less than 0.64%and 0.80%,respectively.The intraday and interday RSDs for L-xylulose were less than 0.31%and 0.59%.The recoveries of xylitol and L-xylulose in fermentation broth were between 99.00%-101.00%.Conclusion There was no interference from other constitutes in the fermentation broth by this method.The methods were suitable for the simultaneous determination of the substrate xylitol and the product L-xylulose in fermentation process.

Simultaneous determination of ten nucleosides and bases contents in plants of Paris L. by HPLC / 中草药

Objective: In order to provide a theory basis for rational development and utilization of medicinal plants from Paris L., this study compared nucleosides (cytidine, uridine, inosine, guanosine, thymidine, adenosine and 2'-desoxyadenossine) and bases (including uracil, guanine, and adenine) contents in the different plants of Paris L.which collected from different places. Methods: RP-HPLC was applied in this study, Venusil MP C18(2) chromatographic column (250 mm × 4.6 mm, 5 μm) was used, the mobile phase was a methanol and water gradient, the flow rate was1.0 mL/min, the temperature of column was 35℃ and the UV detector was set at 260 nm. Sixteen batches of samples in Paris L. were measured by above method, and cluster analysis was used to analyze the results. Results: Ten nucleosides and bases from the plants of Paris L. were measured, a good linear relationship was showed (r2 > 0.999 1), and the average recovery was 96.04%-104.34%, RSD ≤ 2.94%. The samples in Paris L. from different places were divided into three categories by cluster analysis. All of the ten nucleosides and bases were detected in the 16 batches of samples; However, the mass fraction and component ratio of the ten substances were significantly different from each variety. The average of total nucleosides and bases content was ranged from 735.390 1 to 1 612.807 3 μg/g, and the order from high to low was: P. bashanensis > P. polyphylla var. pseudothibetica > P. delavayi var. petiolata > P. axialis > P. polyphylla var. stenophylla > P. polyphylla var. chinensis > P. polyphylla var. yunnanensis > P. marmorata > P. polyphylla var. minor > P. mairei. The mass fraction and component ratio of the ten substances also had a significant difference between the samples collected from different regions which were the same variety. Conclusion: This method is simple, accurate and reliable and has good reproducibility. It is appropriate for measuring the contents of the ten nucleosides and bases. Simultaneously, this study provids a scientific basis for an objective and complete knowledge of the material basis for the efficacy, and for an enrichment and development of evaluation indicators system of in the plants Paris L.

Simultaneous determination of thirteen components in Polygoni Multifori Caulis by UPLC-MS/MS / 中国中药杂志

A comprehensive analytical method based on UPLC-MS/MS was developed for the simultaneous determination of thirteen components including three stilbenes (stilbeneglucoside, polydatin, resveratrol), four anthraquinones (emodin, physcion, emodin-8-β-D-glucopyranoside, aloe-emodin), five flavonoids (epicatechin, rutin, hyperoside, astragalin,quercetin) and one phenolic acid (gallic acid) in Polygoni Multifori Caulis.The separation was carried out on a Waters BEH C18 column（2.1 mm×100 mm,1.7 μm）with gradient elution of acetonitrile-water (0.1% acetic acid) at a flow rate of 0.25 mL•min⁻¹, and column temperature was 35 ℃. The target compounds were analyzed by multiple reaction monitoring (MRM) mode. TOPSIS analysis ware performed to evaluate the samples from different areas and commercial herbs according to the contents of thirteen components. The correlation coefficients of all the calibration curves were higher than 0.991 5. The average recoveries ranged from 95.24% to 102.3%, and the relative standard deviations were less than 5%. The result of TOPSIS analysis showed that the comprehensive quality of Polygoni Multifori Caulis sample from Guangzhou was better. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances, which provided a new basis for the comprehensive assessment and overall control of the quality of Polygoni Multifori Caulis.

Simultaneous determination of 14 organic acids in Shenfu injection by hydrophilic interaction chromatography-tandem mass spectrometry / 中国中药杂志

Organic acids are widely distributed in plants and related products, and participate in a wide range of metabolic pathways (e.g. tricarboxylic acid cycle), showing diverse pharmacological activities. As a widely used Chinese patent medicine, its adverse reactions are often reported. Therefore, we should further clarify the chemical components of Shenfu injection, and prepare strict quality standards to ensure the safety and effectiveness of its clinical use. Shenfu injection is prepared from red ginseng (steamed roots of Panax ginseng) and black prepared lateral roots of Aconitum carmichaelii (Heishunpian) by using modern extraction process, and organic acids are regarded as one of its main components. In current study, a hydrophilic interaction chromatography (HILIC) coupled with mass spectrometric method (HILIC-LC-MS) was developed and validated for the simultaneous determination of 14 organic acids, including cinnamic acid, ferulic acid, 4-hydroxylbenzoic acid, L-(+)-lactic acid, adipic acid, fumaric acid, caffeic acid, succinic acid, maleic acid, malonic acid, D-malic acid, (-)-shikimic acid, D-tartaric acid, and quinic acid in Shenfu injection. Satisfactory retention and separation were achieved for all organic acids on HILIC chromatographic column. Except cinnamic acid (231 μg•L⁻¹), lactic acid (113 μg•L⁻¹) and malonic acid (32.5 μg•L⁻¹), the limit of quantitation for the remaining 11 compounds were less than 10 μg•L⁻¹. D-Malic acid, malonic acid, quinic acid, L-(+)-lactic acid, and cinnamic acid were observed to have higher contents in Shenfu injection (>1.89 mg•L⁻¹), whereas caffeic acid and adipic acid were undetectable in all batches. Above all, the developed method is suitable for the simultaneous determination of organic acids in Shenfu and some other traditional Chinese medicine injections.

Simultaneous Determination of 11 Marker Compounds in Gumiganghwal-tang by HPLC-DAD and LC-MS

Gumiganghwal-tang has been used for the treatment of common cold for a long-time. We developed an accurate and sensitive high performance liquid chromatography-diode array detection (HPLC-DAD) and electrospray ionization mass spectrometry method for the simultaneous determination of ferulic acid, baicalin, bergapten, methyl eugenol, glycyrrhizin, oxypeucedanin, wogonin, nodakenin, atractylenolide III, imperatorin, and atractylenolide I in Gumiganghwal-tang samples. The analytes were separated on a Shiseido C18 column (5 µm, 4.6 mm I.D. × 250 mm) with gradient elution with acetonitrile and 0.1% trifluoroacetic acid. Eleven compounds were quantitatively determined by HPLC-DAD and identified by LC-MS data. We also validated this method. The calibration curves of all the compounds showed good linear regression. The limits of detection and the limits of quantification ranged from 0.04 to 0.63 and from 0.12 to 1.92 µg/mL, respectively. The relative standard deviation values of intra- and inter-days of this method represented less than 2.9%. The recoveries were found to be in the range of 90.06 – 107.66%. The developed method has been successfully applied to the analysis of Gumiganghwal-tang samples. The established HPLC method could be used to quality control of Gumiganghwal-tang.