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Objective:To establish an HPLC-DAD-ELSD method for the simultaneous determination of eight main active components in Buyang Huanwu Decoction, including hydroxysafflor yellow A, paeoniflorin, calycosin glycoside, ferulic acid, ononin, calycosin, fermononetin and astragaloside.Methods:Agilent Eclipse XDB-C18 column (250 mm×4.6 mm, 5 μm) was used with acetonitrile-0.1% formic acid as the mobile phase. The flow rate was 1.0 ml/min; the column temperature was 30 ℃; the detection wavelengths were 230 nm (paeoniflorin), 254 nm (calycosin glycoside, ononin, calycosin, fermononetin), 322 nm (ferulic acid) and 403 nm (hydroxysafflor yellow A); the drift tube temperature of the evaporative light scattering detector was 60 ℃; the carrier gas flow rate was 1.6 L/min.Results:Under these conditions, the separation of hydroxysafflor yellow A, paeoniflorin, calycosin glycoside, ferulic acid, ononin, calycosin, fermononetin and astragaloside was good, and the linear relationship was in line with the requirements ( r=0.994 0-0.999 9). The average recovery was 97.8% - 101.4% ( RSD was 1.28% - 3.70%). Conclusion:The method is simple, stable and reproducible, and can be used for the quality control of Buyang Huanwu Decoction.
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To improve the quality control methods of Poria and develop and utilize its resources fully, alkaline extraction was used in this study to determine the yield and content of alkali-soluble polysaccharides of Poria. The alkali-soluble extracts of Poria were obtained according to the optimum extraction conditions on the basis of single-factor test, and 30 batches of samples were determined. The structure and chemical composition of the alkali-soluble extracts was characterized by high-performance gel permeation chromatography(HPGPC), Fourier transform infrared spectrometry(FT-IR), nuclear magnetic resonance(NMR) spectroscopy and high-performance liquid chromatography(HPLC) with 1-phenyl-3-methyl-5-pyrazolone(PMP-HPLC). The results showed that the content of the alkali-soluble extracts was in the range of 46.98%-73.86%. The main component was β-(1→3)-glucan, and its molecular mass was about 1.093×10~5. Further, the content of alkali-soluble polysaccharides of Poria was measured by UV-Vis spectrophotometry and HPLC coupled with the evaporative light scattering detector(HPLC-ELSD), and 30 batches of samples were measured. The results indicated that the content of alkali-soluble polysaccharides determined by UV-Vis spectrophotometry was in the range of 73.70%-92.57%, and the content of samples from Hubei province was slightly higher than that from Yunnan province, Anhui province and Hunan province. The content of alkali-soluble polysaccharides determined by HPLC-ELSD was in the range of 51.42%-76.69%, and the samples from Hunan province had slightly higher content than that from the other three provinces. The content determined by UV-Vis spectrophotometry was higher than that by HPLC-ELSD. However, the content determined by HPLC-ELSD was close to that of alkali-soluble extract, which could accurately characterize the content of alkali-soluble polysaccharides in Poria, and the method was simple and repeatable. Therefore, it is recommended that the quantitative analysis method for alkali-soluble extract and alkali-soluble polysaccharides by HPLC-ELSD be used in the quality standards of Poria in Chinese Pharmacopeia.
Subject(s)
Poria/chemistry , Spectroscopy, Fourier Transform Infrared , China , Polysaccharides/chemistry , Reference Standards , Chromatography, High Pressure Liquid/methodsABSTRACT
Objective To establish a method for the simultaneous determination of new mangiferin, mangiferin, artemisinin BⅡ, icariin and artemisinin A in Anemarrhenae Rhizoma by high performance liquid chromatography-evaporation light scattering detector (HPLC-ELSD). Methods The column was Agilent Poroshell 120 EC-C18. The mobile phase used acetonitrile-0.2% acetic acid water system with gradient elution. Column temperature was 30 ℃. Flow rate was 0.7 ml/min. Evaporative light scattering detector used nitrogen as atomizing gas. The atomizing gas temperature was 40 ℃ and the drift tube temperature was 90 ℃. The nitrogen volume flow rate was 2.00 L/min and the sample volume was 20 μl. Results The five components were able to achieve baseline separation. Neomangiferin, Mangiferin, Anemaponin BⅡ, Baohuoside I , Anemarrhena saponin AⅢwere determined as 24.1-386 μg/ml (r=0.999 3), 23.2-371 μg/ml (r=0.998 6), 54.2-867.2 μg/ml(r=0.995 6), 5.3-84.8 μg/ml (r=0.996 8), 10-160 μg/ml (r=0.998 9) respectively, which showed a good linear relationship within the concentration range. The average recovery rate of the five components was between 101.8% and 105.0%, and the repeatability RSD was less than 2.4%. The content of the above five components in Zhimu medicinal materials were 1.62%, 0.82%, 7.36%, 0.07%, 0.34%, respectively. Conclusion The method is simple, accurate, and highly sensitive, which could be used as the quantitative determination of multiple index components of Anemarrhenae Rhizoma.
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The aim of this study is to develop the first simultaneous method for quantification of neomycin and polymyxin B inthe presence of dexamethasone using High Performance Liquid Chromatography (HPLC) with an Evaporative LightScattering Detector (ELSD). The analysis was performed using a phenyl Waters X Bridge column, an evaporationtemperature of 50oC, and a nitrogen pressure of 320 kPa. The mobile phase consists of a combination of methanoland trichloroacetate acid (40 mM, pH 1.70–1.80) in gradient mode, flow rate at 1.0 ml/minute, detector gain of 6,and analysis time of 35 minutes. The linearity was achieved with a concentration of 100–500 µg/ml (r = 0.99955)for neomycin and concentration of 30–100 µg/ml (r = 0.99703) for polymyxin B. Recovery results were obtainedbetween 99.150% and 104.773% for neomycin and 96.538% and 105.139% for polymyxin B. The analysis samplefrom the market was found to be 102.27% for neomycin and 100.79% for polymyxin B. The result was compared tothe standard microbiological method. Based on the T-test results of two samples with a 95% confidence level (α =0.05), it was concluded that there was no significant difference between HPLC-ELSD and microbiological methodsfor determining neomycin and polymyxin B. The HPLC-ELSD method has a potential for routine analysis due toadvantages in terms of increasing precision, accuracy, and shorter testing time.
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Evaporative light scattering detector(ELSD) and charged aerosol detector(CAD) methods were established in this study for the content determination of four kinds of sugars in Zhusheyong Yiqi Fumai(YQFM), and the factors affecting the accuracy of CAD methods were discussed. HPLC-ELSD chromatographic separation was performed on a Shodex Asahipak NH2 P-50 column with acetonitrile-water(75∶25)as the mobile phase, with a flow rate of 0.8 mL·min~(-1), drift tube temperature of 80 ℃. The analysis by HPLC-CAD was performed on the same column with acetonitrile-water as mobile phase for gradient elution, with a flow rate of 0.8 mL·min~(-1), a neb temperature of 45 ℃, and power function(PF) of 1.3. The samples of YQFM were detected by ELSD and CAD respectively. It was found that YQFM was composed of fructose, glucose, sucrose and maltose. The linear relationship of the two methods was good, and the recoveries, reproducibility and stability of these four kinds of sugars measured by the two methods satisfied the requirements of methodology. Both CAD and ELSD detectors were accurate and reliable in detecting saccharides components in YQFM. In addition, it was revealed in this study for the first time that the PF parameter of CAD had an important influence on the accuracy of sugar determination and acted as the key parameter of CAD method. It was also found that for CAD, a non-linear detector, there was no significant difference between the results of linear regression and logarithmic regression.
Subject(s)
Aerosols , Carbohydrates , Chromatography, High Pressure Liquid , Light , Reproducibility of Results , Scattering, Radiation , SugarsABSTRACT
Objective To establish the quality standard ofQizhu-Fuzheng Yin, and to conduct a preliminary study on its stability.Methods Corydalis tuber and licorice were identified by TLC. The content of astragaloside was determined by UPLC-ELSD. The initial stability was studied by accelerated test method. Results The spots on TLC plates were clear without interference in the blank reference. The response of astragaloside was linear in the ranges of 36.5-365.0 μg/ml (r2=0.999 2), and the average recovery was 102.8 %, and theRSD was 2.4%. After 1, 2, 3, 6 months tests, the average contents of 3 batches astragaloside were 61.6, 60.4, 60.6μg/ml.ConclusionQizhu-Fuzheng Yin was simple preparation, quality control and stability.
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Objective To summarize the advantage and application of evaporative light scattering detector(ELSD) for the analysis of natural drugs .Methods The application of ELSD in natural medicine analysis was reviewed in the article .Results ELSD in analysis of natural drugs ,natural drugs quality standards research ,fingerprint ,multi‐spectrum combination technology and high‐throughput screening had been widely applied .Conclusion Evaporative light scattering detector had a broad applica‐tion prospect .
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OBJECTIVE: To establish a method for the separation and identification of the components in polysorbate 80 and to lay the foundation for identifing the sensitinogen of polysorbate 80. METHODS: Polysorbate 80 samples from both domestic and foreign manufacturers were collected. The separation condition was first established using HPLC-ELSD, and then the chemical composition of each chromatographic peak was identified by HPLC-MS. RESULTS: The chemical components with different types of structures were efficiently separated, and each chromatographic peak was properly assigned. Polysorbate 80 samples of different batches and manufacturers showed different chemical compositions and relative contents. There was illegal blending during the production of polysorbate 80. CONCLUSION: The proposed method is rapid and simple, which is not only suitable for the process control of polysorbate 80 but also lays foundation for further investigation of the physical and chemical properties of the components thus to find the sensitinogen. Copyright 2012 by the Chinese Pharmaceutical Association.
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A simple and accurate high-performance liquid chromatography（HPLC）coupled with diode array detector（DAD）and evaporative light scattering detector（ELSD）was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation（ZFP）.The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis（HCA）,which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.
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Objective: To determine the contents of celosin A and celosin B in Semen celosiae. Methods: HPLC was carried out with an Agilent Alliance, Model 1100, equipped with Elite ODS C18 column (4.6mm X 250mm, 5 μm) and evaporated light scattering detector (ELSD). The mobile phase (acetonitrile-0.1% glacial acetic acid) was eluted in gradient mode. Results: Celosin A and the peak area showed a good linearship within the range of 2. 5-30.0 μg, with the recovery rate being 99.89% (n=6) and RSD being 1.85%. Celonsin B and the peak area showed a good linearship within the range of 1.25-15.0 μg, with the recovery rate being 98.98% (n=6) and the RSD being 1.78%. Contents of celosin A and celosin B in 8 Semen celosiae samples on medical markets were obviously different from each other, ranging from (0.051 2±0.001 6)%, to (0.114 3 ± 0.000 8)% and (0.010 9±0.001 9)% to (0.081 5±0.000 9)%. Conclusion: We have established an HPLC-ELSD method for determining the effective contents of Semen celosiae; the method is accurate and simple.
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Objective A fingerprint of Rhizoma Alismatis Orientalitis' decoction pieces was established by HPLC-ELSD. Methods The gradient elution mode was applied in chromatographic separation by mobile phase of methanol and water at flow rate of 0.5 mL? min-1 in a Diamonsil TM C18 chromatographic column (250 mm ? 4.6 mm, 5 ? m), and recording time was 70 minute. The parameters of evaporative light scattering detector were set as follows: the temperature of the drift tube was 82.5 ℃ and gas flow rate was 1.9 L ? min-1.The data was analyzed with cluster analysis and similarity calculation software. Results The fingerprint showing 15 common peaks was obtained. The results of cluster analysis and similarity calculation showed there existed differences in fingerprints of 16 batchers of medicinal slices of Rhizoma Alismatis Orientalitis. Conclusion The method is effective and reproducible for the quality control of medicinal slices of Rhizoma Alismatis Orientalitis.