In the present study, a method for the quantitative analysis of multi-components by single marker(QAMS) has been developed and validated for the simultaneous determination of echinacoside(ECH), tubuloside A, acteoside...In the present study, a method for the quantitative analysis of multi-components by single marker(QAMS) has been developed and validated for the simultaneous determination of echinacoside(ECH), tubuloside A, acteoside, isoacteoside, and2’-acetylacteoside in Cistanches Herba. ECH was used as the internal standard(IS) to obtain the relative correction factors(RCFs) of the other four phenylethanoid glycosides(PhGs);meanwhile, various influencing factors on RCFs were investigated under different conditions. The content of each component was calculated with RCF. The results were compared with those obtained by the external standard method(ESM) to verify the feasibility and accuracy of the established QAMS method. No significant difference was found in the quantitative results of 10 batches of Cistanches Herba between QAMS and ESM. The proposed QAMS method for simultaneous determination of PhGs in Cistanches Herba is accurate and feasible, providing an efficient and economical approach for the quality control of Cistanches Herba.展开更多
Cynanchum paniculatum(Bunge) Kitagawa is usually used as an herbal medicine for treating many diseases. Paeonol is the main active component, and its content is the key indicator for quality control of C. paniculatum....Cynanchum paniculatum(Bunge) Kitagawa is usually used as an herbal medicine for treating many diseases. Paeonol is the main active component, and its content is the key indicator for quality control of C. paniculatum. In the present study, we developed a rapid, accurate and precise method for quantitation of paeonol in C. paniculatum using 1 H NMR spectra. The deuterated solvent of methanol-d4 enabled satisfactory separation of the signals to be integrated in 1 H NMR spectrum. H-6(δ 7.78) of 1 H NMR spectrum of C. paniculatum was selected as the feature signal for quantitation, and trimesic acid(TMA) was selected as an internal standard. Validation of the quantitative method was performed in terms of linearity, specificity, repeatability and stability. This is the first time to report quantitative 1 H NMR(qHNMR) applied to determine the content of paeonol in C. paniculatum and showed a wider linearity range than the reported quantitation of paeonol in others. The simple extraction of paeonol from C. paniculatum was rapid and will prompt the application of the developed method. This work implied that qHNMR represented a feasible alternative to HPLC-based methods for quantitation of paeonol in C. paniculatum, and it was suitable for the quality control of C. paniculatum.展开更多
Curcuma wenyujin has been widely used as a traditional medicine in China. In this paper a strategy for the quantitative determination of the polysaccharide by a phenol-sulfuric acid method was described. Involved in t...Curcuma wenyujin has been widely used as a traditional medicine in China. In this paper a strategy for the quantitative determination of the polysaccharide by a phenol-sulfuric acid method was described. Involved in three factors, 5% phenol volume, H2SO4 volume, and temperature of water bath, we adopted the L9（3）3 orthogonal array design to gain the optimal colorimetric method. 3.0 ml of polysaccharide solution, 1.0 ml, 5% phenol and 7.0 ml H2SO4 were mixed with constant stirring in a glass vessel, and then kept in a water bath at 40 ℃. After cooling to room temperature for 20 min, the absorbance values were recorded by the UV-2501 PC spectrometer at the wavelength range of 485 nm. The polysaccharide content in Curcuma wenyujin were 3.21%, 3.23%, 3.20%, 3.18~/0, 3.22% and 2.38%, respectively. All results showed that this method was adequate, valid and applicable, may be applied to the determination of other bacterial polysaccharide as well.展开更多
The click reaction is one of the latest techniques for the functional analysis of bioactive compounds and the analysis makes novel concepts and strategies for medicinal chemistry. N-methylated glycine derivatives have...The click reaction is one of the latest techniques for the functional analysis of bioactive compounds and the analysis makes novel concepts and strategies for medicinal chemistry. N-methylated glycine derivatives have inhibitory activity for the click reaction in direct Cu(I) system because of decrease of Cu(I) concentration. The Cu(I) concentration recovered effectively by sodium ascorbate. Quantitative determination of click reaction at various ligand concentrations revealed that the decrease in reaction yields was observed in a substrate concentration-dependent manner.展开更多
[Objective]This study was to establish a rapid,specific and simple method for quantitative determination of tetrachlorantraniliprole by 1H NMR.[Method]1H NMR spectroscopy was acquired with deuterium DMSO as the solven...[Objective]This study was to establish a rapid,specific and simple method for quantitative determination of tetrachlorantraniliprole by 1H NMR.[Method]1H NMR spectroscopy was acquired with deuterium DMSO as the solvent and maleic acid as internal standard under the conditions of temperature 25℃,pulses width 8.0μs,delay time 5 s,and scanning times 8.[Result]The hydrogen proton peaks of tetrachlorantraniliprole(δ=10.55)and maleic acid(δ=6.27)were taken as quantitative peaks.The peak area ratio y(As/Ar)and mass ratio x(ms/mr)were linearly regressed,and the correlation coefficient was 0.9999.The RSD value of repeatability test was 0.38%,and the RSD value of stability test was 0.77%.The content of tetrachlorantraniliprole was determined as 99.6%.[Conclusion]1H NMR spectroscopy can be used for quantitative determination of tetrachlorantraniliprole without standard reference,which is rapid,accurate and simple.展开更多
It is difficult to identify the source（s） of mixed oils from multiple source rocks, and in particular the relative contribution of each source rock. Artificial mixing experiments using typical crude oils and ratios ...It is difficult to identify the source（s） of mixed oils from multiple source rocks, and in particular the relative contribution of each source rock. Artificial mixing experiments using typical crude oils and ratios of different biomarkers show that the relative contribution changes are non-linear when two oils with different concentrations of biomarkers mix with each other. This may result in an incorrect conclusion if ratios of biomarkers and a simple binary linear equation are used to calculate the contribution proportion of each end-member to the mixed oil. The changes of biomarker ratios with the mixing proportion of end-member oils in the trinal mixing model are more complex than in the binary mixing model. When four or more oils mix, the contribution proportion of each end-member oil to the mixed oil cannot be calculated using biomarker ratios and a simple formula. Artificial mixing experiments on typical oils reveal that the absolute concentrations of biomarkers in the mixed oil cause a linear change with mixing proportion of each end-member. Mathematical inferences verify such linear changes. Some of the mathematical calculation methods using the absolute concentrations or ratios of biomarkers to quantitatively determine the proportion of each end-member in the mixed oils are deduced from the results of artificial experiments and by theoretical inference. Ratio of two biomarker compounds changes as a hyperbola with the mixing proportion in the binary mixing model, as a hyperboloid in the trinal mixing model, and as a hypersurface when mixing more than three end- members. The mixing proportion of each end-member can be quantitatively determined with these mathematical models, using the absolute concentrations and the ratios of biomarkers. The mathematical calculation model is more economical, convenient, accurate and reliable than conventional artificial mixing methods.展开更多
[Objectives]To establish a multi-indicator quality control method for the retention of Longqing Capsule based on the principle of prescription of Chinese medicine.[Methods]High performance liquid chromatography(HPLC)w...[Objectives]To establish a multi-indicator quality control method for the retention of Longqing Capsule based on the principle of prescription of Chinese medicine.[Methods]High performance liquid chromatography(HPLC)with ShimNex CS C 18 as the column;column temperature:35℃;wavelength:270 nm;methanol-0.1%phosphoric acid solution as the mobile phase with gradient elution.[Results]The 12 components of the retention of Longqing Capsule showed good linearity within the investigated range(r≥0.9995),with the average spiked recoveries of 97.83%-100.52%and the RSD of 0.9%-2.1%.[Conclusions]The method is exclusive,sensitive,reproducible,simple and easy to use,and can provide a reference for the construction of the quality standard and control system of Longqing Capsule based on the theory of traditional Chinese medicine.展开更多
[Objectives]To establish an HPLC method for the quantitative determination of multiple phenolic acid components in Tetracera asiatica medicinal material,providing a basis for establishing its quality standards.[Method...[Objectives]To establish an HPLC method for the quantitative determination of multiple phenolic acid components in Tetracera asiatica medicinal material,providing a basis for establishing its quality standards.[Methods]An Inertsil ODS-C 18 column(250 mm×4.6 mm,5μm)was used.The mobile phase consisted of acetonitrile-0.2% phosphoric acid solution(10:90).The flow rate was 1.0 mL/min.The detection wavelength was 274 nm.The column temperature was 25℃.The injection volume was 10μL.The content of three components,gallic acid,protocatechuic acid,and protocatechualdehyde,was determined in 13 batches of T.asiatica.[Results]Gallic acid showed good linearity within the range of 0.020-6.400μg/mL,protocatechuic acid within 0.201-6.432μg/mL,and protocatechualdehyde within 0.202-6.464μg/mL(r>0.9990).The average recovery rates ranged from 98.61%to 101.17%,with RSD s between 1.21%and 2.69%.[Conclusions]The quantitative determination method established in this study is simple and feasible,and can provide a basis for the quality evaluation of T.asiatica.展开更多
In fluorescence flow cytometry,spectral overlap among multiple fluorescent labels cannot be avoided,and thus detected fluorescent intensities need to be compensated.Although fluorescent compensation in flow cytometry ...In fluorescence flow cytometry,spectral overlap among multiple fluorescent labels cannot be avoided,and thus detected fluorescent intensities need to be compensated.Although fluorescent compensation in flow cytometry has been widely used for many years,it still lacks quantitative evaluations to validate its effectiveness.Using a home-developed nine-color fluorescence flow cytometer,this study first obtains calibration curves by assaying gradient concentrations of nine different fluorochromes individually,with the fluorescent intensities of the highest concentrations of each fluorochrome being used to obtain a spillover matrix.Mixed fluorescent solutions are analyzed by flow cytometry in which the obtained fluorescent intensities are compensated by the spillover matrix,translated to specific concentrations based on calibration curve and compared with nominal values.Three mixed solutions of Brilliant Violet 650 and Brilliant Violet 711,of Alexa Fluor 488 and PE,and of Pacific Orange,Alexa Fluor 488,and PE are tested,with fluorescent compensation being observed to reduce excessive signals due to spectral overlap.Specifically,concentration deviations(before vs after compensation)in comparison with nominal values for Brilliant Violet 711 and Alexa Fluor 488 are quantified as 40.6%vs 14.9%and 6.7%vs 1.9%,respectively.The results presented here provide a quantitative reference for fluorescent compensation that can be used to effectively address the issue of spectral overlap in fluorescence flow cytometry.展开更多
Determination of nucleosides and nucleobases is important for the quality control of Fritillaria unibracteata Hsiao et K.C. Hsia var. wabuensis （FUW） due to their physiological and pharmacological actions. In the pr...Determination of nucleosides and nucleobases is important for the quality control of Fritillaria unibracteata Hsiao et K.C. Hsia var. wabuensis （FUW） due to their physiological and pharmacological actions. In the present study, we developed a sensitive and reliable HPLC-diode-array detection method to simultaneously determine ten nucleosides and nucleobases, including cytosine, uracil, cytidine, uridine, thymine, adenine, inosine, guanosine, thymidine and adenosine. Complete separation of all the analytes was achieved on a Zorbax 300 A 300 Extend C18 column with a gradient of methanol-ultrapure water at a flow rate of 1 mL/min in less than 30 min. The diode-array detector wavelength was set at 260 nm for the UV detection of nucleosides and nucleobases. The optimized method provided good linearity （R2〉0.9993 for all the analytes）, satisfactory precision （RSD〈3.715%）, good repeatability （RSD_〈3.748%） and good recovery （RSD from 97.688% to 102.923%）. In addition, the developed method was successfully applied to simultaneous determination of ten nucleosides and nucleobases from FUW, and their content changes of various cultivation time （1-7 years） were further analyzed for the first time. Our findings were useful for ensuring the cultivation time choice of artificial cultivation, quality control, pharmaceutical studies and clinical efficacy of FUW.展开更多
In this study, we developed a novel and simple HPLC-DAD method for simultaneous qualitative and quantitative determination of five major components of Glycyrrhizae Radix et Rhizoma （GRR） in Xiaoer Zhike Tangjiang （...In this study, we developed a novel and simple HPLC-DAD method for simultaneous qualitative and quantitative determination of five major components of Glycyrrhizae Radix et Rhizoma （GRR） in Xiaoer Zhike Tangjiang （XEZKTJ） with standardized reference extract （SRE）. The five analytes （liquiritin apioside, liquiritin, isoliquiritin apioside, liquirigenin and glycyrrhizic acid） were well separated with good linearity, precision, stability and repeatability. The recovery rates ranged from 95.69% to 100.80%. The content of the five compounds in 34 batches of commercial XEZKTJ products was determined using standardized GRR extract （SRE method） and individual chemical reference standards （CRS method）. Highly similar results were obtained from the two methods, demonstrating the feasibility of the proposed SRE method. Taken together, we proposed an efficient and low-cost way to perform multi-component quality control of XEZKTJ in this study.展开更多
A reverse-phase HPLC method was developed for the simultaneous separation and determination of five bioactive phenolic acids,yunnaneic acid E,rosmarinic acid,lithospermic acid,salvianolic acid B and salvianolic acid A...A reverse-phase HPLC method was developed for the simultaneous separation and determination of five bioactive phenolic acids,yunnaneic acid E,rosmarinic acid,lithospermic acid,salvianolic acid B and salvianolic acid A in eight different samples of Salvia yunnanensis collected in Yunnan Province.For comparison,the sample of Salvia miltiorrhiza was included. All the samples were extracted for 60 min with 50%methanol in an ultrasonic bath.The optimal separation was achieved on a YMC-Pack Pro C18 column,with a gradient of 0.1%（v/v） phosphoric acid and acetonitrile,at a flow rate of 1.0 mL/min and at a detection wavelength of 280 nm.The separation was obtained within 65 min for five bioactive phenolic acids.All calibration curves showed good linearity（r^2〉0.999） within test ranges.The relative standard deviation of the method was less than 5%for intra- and inter-day assays.The mean recovery of the method was in the range from 97%to 104%,with RSD less than 5%.This assay was successfully applied to the quantitative determination of five bioactive phenolic acids in nine resource samples. The results showed that the developed HPLC assay was suitable for the quality control of S.yunnanensis and it can be used to differentiate S.yunnanensis from S.miltiorrhiza.展开更多
In this paper, a sensitive and specific liquid chromatography-electrospray ionisation-mass spectrometry （LC-ESI-MS） method has been developed and validated for the identification and determination of seven flavonoid...In this paper, a sensitive and specific liquid chromatography-electrospray ionisation-mass spectrometry （LC-ESI-MS） method has been developed and validated for the identification and determination of seven flavonoids, namely epimedin A, epimedin B, epimedin C, icariin, sagittatoside B, 2＂-O-rhamnosyl icariside II, and baohuoside I in Epimedium from different sources.展开更多
Si-Mo-Tang（SMT） oral liquid preparation, a traditional Chinese medicine, was prepared from four crude herbal drugs, Fructus Aurantii Submaturus, Radix Aucklandiae, Semen Arecae and Radix Linderae Aggregatae. A combi...Si-Mo-Tang（SMT） oral liquid preparation, a traditional Chinese medicine, was prepared from four crude herbal drugs, Fructus Aurantii Submaturus, Radix Aucklandiae, Semen Arecae and Radix Linderae Aggregatae. A combinative method using HPLC fingerprint and quantitative analysis was developed and validated for quality consistency evaluation of SMT. Individual HPLC chromatograms were evaluated against the mean chromatogram generated via a similarity evaluation computer program. Data from chromatographic fingerprints were also processed with principal component analysis（PCA） and hierarchical cluster analysis（HCA）. Additionally, six components （naringin, isonaringin, hesperidin, neohesperidin, norisoboldine and potassium sorbate） in SMT were simultaneously determined to interpret the quality consistency. For fingerprint analysis, 20 peaks were selected as the characteristic peaks to evaluate the similarities of 26 SMT collected from different manufacturers. Among the 20 characteristic peaks, 10 peaks were assigned to be naringin, hesperidin, neohesperidin, isonaringin, neoeriocitrin, tangeretin, nobiletin, norisoboldine, 5-（ethoxymethyl）furan-2-carbaldehyde and potassium sorbate, respectively. The results of similarity analysis, PCA and HCA, indicate that the samples from different manufacturers were consistent with each other in composition. The results from the quantitative data show that the contents of six compounds were significantly different in SMT oral liquid preparations from different manufacturers. The combinative method of chromatographic fingerprint with quantitative analysis developed here offered an efficient way for the quality consistency evaluation of the traditional Chinese medicine SMT.展开更多
Aconiti Lateralis Radix Praeparata(Fuzi) is a commonly used traditional Chinese medicine in clinic for its potency in restoring yang and rescuing from collapse. Aconiti alkaloids, mainly including monoester-diterpenoi...Aconiti Lateralis Radix Praeparata(Fuzi) is a commonly used traditional Chinese medicine in clinic for its potency in restoring yang and rescuing from collapse. Aconiti alkaloids, mainly including monoester-diterpenoidaconitines(MDAs) and diester-diterpenoidaconitines(DDAs), are considered to act as both bioactive and toxic constituents. In the present study, a feasible, economical, and accurate HPLC method for simultaneous determination of six alkaloid markers using the Single Standard for Determination of Multi-Components(SSDMC) method was developed and fully validated. Benzoylmesaconine was used as the unique reference standard. This method was proven as accurate(recovery varying between 97.5%-101.8%, RSD < 3%), precise(RSD 0.63%-2.05%), and linear(R > 0.999 9) over the concentration ranges, and subsequently applied to quantitative evaluation of 62 batches of samples, among which 45 batches were from good manufacturing practice(GMP) facilities and 17 batches from the drug market. The contents were then analyzed by principal component analysis(PCA) and homogeneity test. The present study provided valuable information for improving the quality standard of Aconiti Lateralis Radix Praeparata. The developed method also has the potential in analysis of other Aconitum species, such as Aconitum carmichaelii(prepared parent root) and Aconitum kusnezoffii(prepared root).展开更多
To develop an HPLC-ELSD method for the determination of nine inulin-type fructo-oligosaccharides in inulin,the HPLC-ELSD system consisted of Waters XBridge■ Amide column(4.6 mm×250 mm,5μm)with a gradient elutio...To develop an HPLC-ELSD method for the determination of nine inulin-type fructo-oligosaccharides in inulin,the HPLC-ELSD system consisted of Waters XBridge■ Amide column(4.6 mm×250 mm,5μm)with a gradient elution mobile phase consisting of acetonitrile and water at a flow rate of 1.2 mL/min at 30°C.The detector was an Agilent Technologies 380-ELSD.The drift tube temperature for the ELSD was set at 55°C with a nitrogen flow rate of 1.8 L/min.The injection volume was 15μL.The results showed that the detection range for the nine inulin-type fructo-oligosaccharides was 3.81–30.60μg R^(2)=0.99969 for kestose,3.73–29.97μg R^(2)=0.99981 for nystose,3.82–30.69μg R^(2)=0.99993 for fructosylnystose,3.80–30.48μg R^(2)=0.99995 for GF5,3.73–29.96μg R^(2)=0.99993 for GF6,3.78–30.30μg R^(2)=0.99983 for GF7,3.82–30μg R^(2)=0.99989 for GF8,3.71–29.80μg R^(2)=0.99974 for GF9,3.61–29.00μg R^(2)=0.99970 for GF10,respectively.The recovery of the nine oligosaccharides ranged between 96.48%–100.84%(n=6).The method was simple,accurate,and reproducible that it could be used as an analytical method for evaluating the quality of inulin effectively.展开更多
We present here the application of one-dimensional and two-dimensional NMR techniques to characterize the structure of methoxyl end-functionalized polystyrenes （PS）. The peaks in 1H-NMR spectra corresponding to main...We present here the application of one-dimensional and two-dimensional NMR techniques to characterize the structure of methoxyl end-functionalized polystyrenes （PS）. The peaks in 1H-NMR spectra corresponding to main-chain, side-chain and chain-end groups are assigned by 1H-1H gCOSY, 1H-13C gHSQC and gHMBC spectra. For the first time, the spin-lattice relaxation time （T1） of protons of the chain-ends is revealed to be affected more by polymer molecular weight （MW） than by the protons of the main-chains and the side-chains （almost independent from MW）. As a result, a much higher delay time （dl） for chain-ends （d1〉 20T1） is needed for quantitative NMR measurement when using end-group estimation method to obtain the MW of PS, which is in accordance with the value estimated by GPC. An improved method for the polymer MW determination is established, by combination of different NMR techniques to distinguish the peaks, and a large dl setting to achieve quantitative NMR analysis.展开更多
Objective:To establish an high performance liquid chromatography-quantitative analysis of multi components by single marker(HPLC-QAMS)method for simultaneous determination of gastrodin,parishin E,parishin B,parishin,t...Objective:To establish an high performance liquid chromatography-quantitative analysis of multi components by single marker(HPLC-QAMS)method for simultaneous determination of gastrodin,parishin E,parishin B,parishin,tenuifolin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andαasarone in Dianxiankang Capsules.Methods:Waters Symmetry C_(18)column was used with acetonitrile-0.05%phosphoric acid solution as mobile phase for gradient elution.Multiwavelength switching detection.The contents of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were calculated by relative correction factor.At the same time,the contents of 10 components in 12 batches of Dianxiankang Capsules were determined by external standard method(ESM).Results:An HPLC-QAMS method was established tenuifolin as the internal reference substance was established.The relative correction factors of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were 0.8238,0.7239,1.0229,1.1881,0.7272,1.3108,0.9314,0.6549 and 1.0572,respectively.The relative correction factors had good repeatability and no significant difference with ESM(P>0.05).Conclusion:HPLC-QAMS can be used for simultaneous determination of multi-index components in Dianxiankang Capsules.展开更多
[Objectives]This study was conducted to establish a method for the simultaneous determination of caffeic acid, rutin, ononin, luteolin, and apigenin in Operculina turpethum(L.) S. Manso. [Methods]With ononin from O. t...[Objectives]This study was conducted to establish a method for the simultaneous determination of caffeic acid, rutin, ononin, luteolin, and apigenin in Operculina turpethum(L.) S. Manso. [Methods]With ononin from O. turpethum as the internal reference, the five components were separated by HPLC, and the contents of various components were calculated according to the relative correction factors of ononin with caffeic acid, rutin, luteolin, and apigenin. Meanwhile, the calculated results of quantitative analysis of multi-components by single marker(QAMS) were compared with the determined values of the external standard method. [Results] The linear relationship of the five components in their respective ranges was good(r=0.999 9). The average recovery was in the range of 97.48%-101.05%, and the RSD values were in the range of 1.04%-2.71%. The results obtained by QAMS were close to those obtained by the external standard method. [Conclusions] The method is accurate, stable and adaptable, and can be used for the determination of five flavonoids in O. turpethum.展开更多
基金National Key R&D Program of China(Grant Nos.2017YFC1702400,2018YFC1707300 and 2018YFC1707904)
文摘In the present study, a method for the quantitative analysis of multi-components by single marker(QAMS) has been developed and validated for the simultaneous determination of echinacoside(ECH), tubuloside A, acteoside, isoacteoside, and2’-acetylacteoside in Cistanches Herba. ECH was used as the internal standard(IS) to obtain the relative correction factors(RCFs) of the other four phenylethanoid glycosides(PhGs);meanwhile, various influencing factors on RCFs were investigated under different conditions. The content of each component was calculated with RCF. The results were compared with those obtained by the external standard method(ESM) to verify the feasibility and accuracy of the established QAMS method. No significant difference was found in the quantitative results of 10 batches of Cistanches Herba between QAMS and ESM. The proposed QAMS method for simultaneous determination of PhGs in Cistanches Herba is accurate and feasible, providing an efficient and economical approach for the quality control of Cistanches Herba.
基金Science and Technology Project of Yunnan Provincial Department of Science and Technology(Grant No.2018FD081)Yunnan Local Colleges Applied Basic Research Projects[Grant Nos.2019FH001-(109),2017FH001-092,2017FH001-094 and 2018FH001-019]Youth Research Foundation of Qujing Normal University in 2019(Grant No.2019QN004)。
文摘Cynanchum paniculatum(Bunge) Kitagawa is usually used as an herbal medicine for treating many diseases. Paeonol is the main active component, and its content is the key indicator for quality control of C. paniculatum. In the present study, we developed a rapid, accurate and precise method for quantitation of paeonol in C. paniculatum using 1 H NMR spectra. The deuterated solvent of methanol-d4 enabled satisfactory separation of the signals to be integrated in 1 H NMR spectrum. H-6(δ 7.78) of 1 H NMR spectrum of C. paniculatum was selected as the feature signal for quantitation, and trimesic acid(TMA) was selected as an internal standard. Validation of the quantitative method was performed in terms of linearity, specificity, repeatability and stability. This is the first time to report quantitative 1 H NMR(qHNMR) applied to determine the content of paeonol in C. paniculatum and showed a wider linearity range than the reported quantitation of paeonol in others. The simple extraction of paeonol from C. paniculatum was rapid and will prompt the application of the developed method. This work implied that qHNMR represented a feasible alternative to HPLC-based methods for quantitation of paeonol in C. paniculatum, and it was suitable for the quality control of C. paniculatum.
文摘Curcuma wenyujin has been widely used as a traditional medicine in China. In this paper a strategy for the quantitative determination of the polysaccharide by a phenol-sulfuric acid method was described. Involved in three factors, 5% phenol volume, H2SO4 volume, and temperature of water bath, we adopted the L9（3）3 orthogonal array design to gain the optimal colorimetric method. 3.0 ml of polysaccharide solution, 1.0 ml, 5% phenol and 7.0 ml H2SO4 were mixed with constant stirring in a glass vessel, and then kept in a water bath at 40 ℃. After cooling to room temperature for 20 min, the absorbance values were recorded by the UV-2501 PC spectrometer at the wavelength range of 485 nm. The polysaccharide content in Curcuma wenyujin were 3.21%, 3.23%, 3.20%, 3.18~/0, 3.22% and 2.38%, respectively. All results showed that this method was adequate, valid and applicable, may be applied to the determination of other bacterial polysaccharide as well.
文摘The click reaction is one of the latest techniques for the functional analysis of bioactive compounds and the analysis makes novel concepts and strategies for medicinal chemistry. N-methylated glycine derivatives have inhibitory activity for the click reaction in direct Cu(I) system because of decrease of Cu(I) concentration. The Cu(I) concentration recovered effectively by sodium ascorbate. Quantitative determination of click reaction at various ligand concentrations revealed that the decrease in reaction yields was observed in a substrate concentration-dependent manner.
文摘[Objective]This study was to establish a rapid,specific and simple method for quantitative determination of tetrachlorantraniliprole by 1H NMR.[Method]1H NMR spectroscopy was acquired with deuterium DMSO as the solvent and maleic acid as internal standard under the conditions of temperature 25℃,pulses width 8.0μs,delay time 5 s,and scanning times 8.[Result]The hydrogen proton peaks of tetrachlorantraniliprole(δ=10.55)and maleic acid(δ=6.27)were taken as quantitative peaks.The peak area ratio y(As/Ar)and mass ratio x(ms/mr)were linearly regressed,and the correlation coefficient was 0.9999.The RSD value of repeatability test was 0.38%,and the RSD value of stability test was 0.77%.The content of tetrachlorantraniliprole was determined as 99.6%.[Conclusion]1H NMR spectroscopy can be used for quantitative determination of tetrachlorantraniliprole without standard reference,which is rapid,accurate and simple.
文摘It is difficult to identify the source（s） of mixed oils from multiple source rocks, and in particular the relative contribution of each source rock. Artificial mixing experiments using typical crude oils and ratios of different biomarkers show that the relative contribution changes are non-linear when two oils with different concentrations of biomarkers mix with each other. This may result in an incorrect conclusion if ratios of biomarkers and a simple binary linear equation are used to calculate the contribution proportion of each end-member to the mixed oil. The changes of biomarker ratios with the mixing proportion of end-member oils in the trinal mixing model are more complex than in the binary mixing model. When four or more oils mix, the contribution proportion of each end-member oil to the mixed oil cannot be calculated using biomarker ratios and a simple formula. Artificial mixing experiments on typical oils reveal that the absolute concentrations of biomarkers in the mixed oil cause a linear change with mixing proportion of each end-member. Mathematical inferences verify such linear changes. Some of the mathematical calculation methods using the absolute concentrations or ratios of biomarkers to quantitatively determine the proportion of each end-member in the mixed oils are deduced from the results of artificial experiments and by theoretical inference. Ratio of two biomarker compounds changes as a hyperbola with the mixing proportion in the binary mixing model, as a hyperboloid in the trinal mixing model, and as a hypersurface when mixing more than three end- members. The mixing proportion of each end-member can be quantitatively determined with these mathematical models, using the absolute concentrations and the ratios of biomarkers. The mathematical calculation model is more economical, convenient, accurate and reliable than conventional artificial mixing methods.
基金Supported by Provincial University Scientific Research Platform Team Project of Guizhou Provincial Department of Education(Qianjiaoji[2022]No.010).
文摘[Objectives]To establish a multi-indicator quality control method for the retention of Longqing Capsule based on the principle of prescription of Chinese medicine.[Methods]High performance liquid chromatography(HPLC)with ShimNex CS C 18 as the column;column temperature:35℃;wavelength:270 nm;methanol-0.1%phosphoric acid solution as the mobile phase with gradient elution.[Results]The 12 components of the retention of Longqing Capsule showed good linearity within the investigated range(r≥0.9995),with the average spiked recoveries of 97.83%-100.52%and the RSD of 0.9%-2.1%.[Conclusions]The method is exclusive,sensitive,reproducible,simple and easy to use,and can provide a reference for the construction of the quality standard and control system of Longqing Capsule based on the theory of traditional Chinese medicine.
基金Supported by Regional Science Foundation of China,National Natural Science Foundation(No.82160820)General Program of Guizhou Provincial Natural Science Foundation[QianKeHe Foundation-ZK(2023)General153].
文摘[Objectives]To establish an HPLC method for the quantitative determination of multiple phenolic acid components in Tetracera asiatica medicinal material,providing a basis for establishing its quality standards.[Methods]An Inertsil ODS-C 18 column(250 mm×4.6 mm,5μm)was used.The mobile phase consisted of acetonitrile-0.2% phosphoric acid solution(10:90).The flow rate was 1.0 mL/min.The detection wavelength was 274 nm.The column temperature was 25℃.The injection volume was 10μL.The content of three components,gallic acid,protocatechuic acid,and protocatechualdehyde,was determined in 13 batches of T.asiatica.[Results]Gallic acid showed good linearity within the range of 0.020-6.400μg/mL,protocatechuic acid within 0.201-6.432μg/mL,and protocatechualdehyde within 0.202-6.464μg/mL(r>0.9990).The average recovery rates ranged from 98.61%to 101.17%,with RSD s between 1.21%and 2.69%.[Conclusions]The quantitative determination method established in this study is simple and feasible,and can provide a basis for the quality evaluation of T.asiatica.
基金support from the National Natural Science Foundation of China(Grant Nos.62331025 and 62121003).
文摘In fluorescence flow cytometry,spectral overlap among multiple fluorescent labels cannot be avoided,and thus detected fluorescent intensities need to be compensated.Although fluorescent compensation in flow cytometry has been widely used for many years,it still lacks quantitative evaluations to validate its effectiveness.Using a home-developed nine-color fluorescence flow cytometer,this study first obtains calibration curves by assaying gradient concentrations of nine different fluorochromes individually,with the fluorescent intensities of the highest concentrations of each fluorochrome being used to obtain a spillover matrix.Mixed fluorescent solutions are analyzed by flow cytometry in which the obtained fluorescent intensities are compensated by the spillover matrix,translated to specific concentrations based on calibration curve and compared with nominal values.Three mixed solutions of Brilliant Violet 650 and Brilliant Violet 711,of Alexa Fluor 488 and PE,and of Pacific Orange,Alexa Fluor 488,and PE are tested,with fluorescent compensation being observed to reduce excessive signals due to spectral overlap.Specifically,concentration deviations(before vs after compensation)in comparison with nominal values for Brilliant Violet 711 and Alexa Fluor 488 are quantified as 40.6%vs 14.9%and 6.7%vs 1.9%,respectively.The results presented here provide a quantitative reference for fluorescent compensation that can be used to effectively address the issue of spectral overlap in fluorescence flow cytometry.
基金The Specialized Research Fund for the Doctoral Program of Higher Education of China(Grant No.20115103110009)"211"Project Double-Support Plan of Sichuan Agricultural Un iversity(Grant No.03570313)Modernization of Chinese Traditional Medicines in Hainan Province(Grant No.ZY201410)
文摘Determination of nucleosides and nucleobases is important for the quality control of Fritillaria unibracteata Hsiao et K.C. Hsia var. wabuensis （FUW） due to their physiological and pharmacological actions. In the present study, we developed a sensitive and reliable HPLC-diode-array detection method to simultaneously determine ten nucleosides and nucleobases, including cytosine, uracil, cytidine, uridine, thymine, adenine, inosine, guanosine, thymidine and adenosine. Complete separation of all the analytes was achieved on a Zorbax 300 A 300 Extend C18 column with a gradient of methanol-ultrapure water at a flow rate of 1 mL/min in less than 30 min. The diode-array detector wavelength was set at 260 nm for the UV detection of nucleosides and nucleobases. The optimized method provided good linearity （R2〉0.9993 for all the analytes）, satisfactory precision （RSD〈3.715%）, good repeatability （RSD_〈3.748%） and good recovery （RSD from 97.688% to 102.923%）. In addition, the developed method was successfully applied to simultaneous determination of ten nucleosides and nucleobases from FUW, and their content changes of various cultivation time （1-7 years） were further analyzed for the first time. Our findings were useful for ensuring the cultivation time choice of artificial cultivation, quality control, pharmaceutical studies and clinical efficacy of FUW.
基金Drug Standards Improvement Project of Chinese Pharmacopoeia Commission
文摘In this study, we developed a novel and simple HPLC-DAD method for simultaneous qualitative and quantitative determination of five major components of Glycyrrhizae Radix et Rhizoma （GRR） in Xiaoer Zhike Tangjiang （XEZKTJ） with standardized reference extract （SRE）. The five analytes （liquiritin apioside, liquiritin, isoliquiritin apioside, liquirigenin and glycyrrhizic acid） were well separated with good linearity, precision, stability and repeatability. The recovery rates ranged from 95.69% to 100.80%. The content of the five compounds in 34 batches of commercial XEZKTJ products was determined using standardized GRR extract （SRE method） and individual chemical reference standards （CRS method）. Highly similar results were obtained from the two methods, demonstrating the feasibility of the proposed SRE method. Taken together, we proposed an efficient and low-cost way to perform multi-component quality control of XEZKTJ in this study.
基金National High-Tech R & D Program(863 Program, Grant No.2004AA2Z3342)
文摘A reverse-phase HPLC method was developed for the simultaneous separation and determination of five bioactive phenolic acids,yunnaneic acid E,rosmarinic acid,lithospermic acid,salvianolic acid B and salvianolic acid A in eight different samples of Salvia yunnanensis collected in Yunnan Province.For comparison,the sample of Salvia miltiorrhiza was included. All the samples were extracted for 60 min with 50%methanol in an ultrasonic bath.The optimal separation was achieved on a YMC-Pack Pro C18 column,with a gradient of 0.1%（v/v） phosphoric acid and acetonitrile,at a flow rate of 1.0 mL/min and at a detection wavelength of 280 nm.The separation was obtained within 65 min for five bioactive phenolic acids.All calibration curves showed good linearity（r^2〉0.999） within test ranges.The relative standard deviation of the method was less than 5%for intra- and inter-day assays.The mean recovery of the method was in the range from 97%to 104%,with RSD less than 5%.This assay was successfully applied to the quantitative determination of five bioactive phenolic acids in nine resource samples. The results showed that the developed HPLC assay was suitable for the quality control of S.yunnanensis and it can be used to differentiate S.yunnanensis from S.miltiorrhiza.
文摘In this paper, a sensitive and specific liquid chromatography-electrospray ionisation-mass spectrometry （LC-ESI-MS） method has been developed and validated for the identification and determination of seven flavonoids, namely epimedin A, epimedin B, epimedin C, icariin, sagittatoside B, 2＂-O-rhamnosyl icariside II, and baohuoside I in Epimedium from different sources.
基金Supported by the National Basic Research Program of China(No.2009CB523002)the National Action of Technology Personnel Servicing Enterprise Program of China(No.2009FJ5049)+1 种基金the Foundation of Hunan Science and Technology Committee, China(No.2009XK6032, 2009-152)the Foundation of Hunan Educational Committee, China(No.09CY001)
文摘Si-Mo-Tang（SMT） oral liquid preparation, a traditional Chinese medicine, was prepared from four crude herbal drugs, Fructus Aurantii Submaturus, Radix Aucklandiae, Semen Arecae and Radix Linderae Aggregatae. A combinative method using HPLC fingerprint and quantitative analysis was developed and validated for quality consistency evaluation of SMT. Individual HPLC chromatograms were evaluated against the mean chromatogram generated via a similarity evaluation computer program. Data from chromatographic fingerprints were also processed with principal component analysis（PCA） and hierarchical cluster analysis（HCA）. Additionally, six components （naringin, isonaringin, hesperidin, neohesperidin, norisoboldine and potassium sorbate） in SMT were simultaneously determined to interpret the quality consistency. For fingerprint analysis, 20 peaks were selected as the characteristic peaks to evaluate the similarities of 26 SMT collected from different manufacturers. Among the 20 characteristic peaks, 10 peaks were assigned to be naringin, hesperidin, neohesperidin, isonaringin, neoeriocitrin, tangeretin, nobiletin, norisoboldine, 5-（ethoxymethyl）furan-2-carbaldehyde and potassium sorbate, respectively. The results of similarity analysis, PCA and HCA, indicate that the samples from different manufacturers were consistent with each other in composition. The results from the quantitative data show that the contents of six compounds were significantly different in SMT oral liquid preparations from different manufacturers. The combinative method of chromatographic fingerprint with quantitative analysis developed here offered an efficient way for the quality consistency evaluation of the traditional Chinese medicine SMT.
基金financially supported by the project of Standardization of Traditional Chinese Medicines/Indigenous Drugs sponsored by Chinese Academy of Sciences(No.KSZD-EW-Z-004-01)
文摘Aconiti Lateralis Radix Praeparata(Fuzi) is a commonly used traditional Chinese medicine in clinic for its potency in restoring yang and rescuing from collapse. Aconiti alkaloids, mainly including monoester-diterpenoidaconitines(MDAs) and diester-diterpenoidaconitines(DDAs), are considered to act as both bioactive and toxic constituents. In the present study, a feasible, economical, and accurate HPLC method for simultaneous determination of six alkaloid markers using the Single Standard for Determination of Multi-Components(SSDMC) method was developed and fully validated. Benzoylmesaconine was used as the unique reference standard. This method was proven as accurate(recovery varying between 97.5%-101.8%, RSD < 3%), precise(RSD 0.63%-2.05%), and linear(R > 0.999 9) over the concentration ranges, and subsequently applied to quantitative evaluation of 62 batches of samples, among which 45 batches were from good manufacturing practice(GMP) facilities and 17 batches from the drug market. The contents were then analyzed by principal component analysis(PCA) and homogeneity test. The present study provided valuable information for improving the quality standard of Aconiti Lateralis Radix Praeparata. The developed method also has the potential in analysis of other Aconitum species, such as Aconitum carmichaelii(prepared parent root) and Aconitum kusnezoffii(prepared root).
基金National Natural Science Foundation of China(Grant No.21977005)the Beijing Natural Science Foundation(Grant No.7192101)。
文摘To develop an HPLC-ELSD method for the determination of nine inulin-type fructo-oligosaccharides in inulin,the HPLC-ELSD system consisted of Waters XBridge■ Amide column(4.6 mm×250 mm,5μm)with a gradient elution mobile phase consisting of acetonitrile and water at a flow rate of 1.2 mL/min at 30°C.The detector was an Agilent Technologies 380-ELSD.The drift tube temperature for the ELSD was set at 55°C with a nitrogen flow rate of 1.8 L/min.The injection volume was 15μL.The results showed that the detection range for the nine inulin-type fructo-oligosaccharides was 3.81–30.60μg R^(2)=0.99969 for kestose,3.73–29.97μg R^(2)=0.99981 for nystose,3.82–30.69μg R^(2)=0.99993 for fructosylnystose,3.80–30.48μg R^(2)=0.99995 for GF5,3.73–29.96μg R^(2)=0.99993 for GF6,3.78–30.30μg R^(2)=0.99983 for GF7,3.82–30μg R^(2)=0.99989 for GF8,3.71–29.80μg R^(2)=0.99974 for GF9,3.61–29.00μg R^(2)=0.99970 for GF10,respectively.The recovery of the nine oligosaccharides ranged between 96.48%–100.84%(n=6).The method was simple,accurate,and reproducible that it could be used as an analytical method for evaluating the quality of inulin effectively.
基金supported by the National Natural Science Foundation of China(Nos.21274099,21305098 and 21474067)the Priority Academic Program Development of Jiangsu High Education Institutions
文摘We present here the application of one-dimensional and two-dimensional NMR techniques to characterize the structure of methoxyl end-functionalized polystyrenes （PS）. The peaks in 1H-NMR spectra corresponding to main-chain, side-chain and chain-end groups are assigned by 1H-1H gCOSY, 1H-13C gHSQC and gHMBC spectra. For the first time, the spin-lattice relaxation time （T1） of protons of the chain-ends is revealed to be affected more by polymer molecular weight （MW） than by the protons of the main-chains and the side-chains （almost independent from MW）. As a result, a much higher delay time （dl） for chain-ends （d1〉 20T1） is needed for quantitative NMR measurement when using end-group estimation method to obtain the MW of PS, which is in accordance with the value estimated by GPC. An improved method for the polymer MW determination is established, by combination of different NMR techniques to distinguish the peaks, and a large dl setting to achieve quantitative NMR analysis.
基金National Traditional Chinese Medicine Technology Inheritance Talent Training Project(NO.T20194828003)。
文摘Objective:To establish an high performance liquid chromatography-quantitative analysis of multi components by single marker(HPLC-QAMS)method for simultaneous determination of gastrodin,parishin E,parishin B,parishin,tenuifolin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andαasarone in Dianxiankang Capsules.Methods:Waters Symmetry C_(18)column was used with acetonitrile-0.05%phosphoric acid solution as mobile phase for gradient elution.Multiwavelength switching detection.The contents of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were calculated by relative correction factor.At the same time,the contents of 10 components in 12 batches of Dianxiankang Capsules were determined by external standard method(ESM).Results:An HPLC-QAMS method was established tenuifolin as the internal reference substance was established.The relative correction factors of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were 0.8238,0.7239,1.0229,1.1881,0.7272,1.3108,0.9314,0.6549 and 1.0572,respectively.The relative correction factors had good repeatability and no significant difference with ESM(P>0.05).Conclusion:HPLC-QAMS can be used for simultaneous determination of multi-index components in Dianxiankang Capsules.
基金Supported by Guangxi Natural Science Foundation (2018GXNSFAA281138,2022JJA140749)Open Project for the Construction of First-class Disciplines in Guangxi (2019XK134)Key Laboratory of Extraction,Purification and Quality Analysis of Traditional Chinese Medicine in Colleges and Universities of Guangxi(GJKY[2014]6)。
文摘[Objectives]This study was conducted to establish a method for the simultaneous determination of caffeic acid, rutin, ononin, luteolin, and apigenin in Operculina turpethum(L.) S. Manso. [Methods]With ononin from O. turpethum as the internal reference, the five components were separated by HPLC, and the contents of various components were calculated according to the relative correction factors of ononin with caffeic acid, rutin, luteolin, and apigenin. Meanwhile, the calculated results of quantitative analysis of multi-components by single marker(QAMS) were compared with the determined values of the external standard method. [Results] The linear relationship of the five components in their respective ranges was good(r=0.999 9). The average recovery was in the range of 97.48%-101.05%, and the RSD values were in the range of 1.04%-2.71%. The results obtained by QAMS were close to those obtained by the external standard method. [Conclusions] The method is accurate, stable and adaptable, and can be used for the determination of five flavonoids in O. turpethum.
