[Objective] This study aimed to establish a pre-column derivatization HPLC method for the identification and analysis of monosaccharide composition of Pu-erh tea polysaccharide. [Method] Pu-erh tea polysaccharide was ...[Objective] This study aimed to establish a pre-column derivatization HPLC method for the identification and analysis of monosaccharide composition of Pu-erh tea polysaccharide. [Method] Pu-erh tea polysaccharide was extracted using the wa- ter extraction method, further isolated and purified by DEAE cellulose-52 columns. The obtained tea polysaccharide and four components TPS1, TPS2, TPS3 and TPS, were first derived by 1-phenyl-3-methyl-5-pyrazolone （PMP）, and then the PMP derivatives of monosaccharide were analyzed by high performance liquid chromatog- raphy （HPLC）. [Result] Pu-erh tea polysaccharide contained eight kinds of monosac- chaddes （mannose, rhamnose, glucuronic acid, galacturonic acid, grucose, galactose, arabinose, fucose）, without xylose; so it was the same with TPS1; each of TPS2, TPS3 and TPS4 contained seven monosaccharides, while no fucose. [Conclusion] This method is simplified and rapid, which can be used to determine the monosac- charide composition of Pu-erh tea polysaccharide and monosaccharide content.展开更多
A rapid and accurate quantitative method of high performance liquid chromatography( HPLC) with fluorescence detector has been developed for the analysis of 18 kinds of amino acids in fresh tea leaves. The samples were...A rapid and accurate quantitative method of high performance liquid chromatography( HPLC) with fluorescence detector has been developed for the analysis of 18 kinds of amino acids in fresh tea leaves. The samples were minced and mixed,and extracted with ultra pure water at 90℃ for 20 min. The 6-aminoquinolyl N-hydroxy-succinimidyl carbamate( AQC) was used as pre-column derivatization reagent. Gradient HPLC separation was performed on a C_(18) column( Symmetry C_(18),3. 9 mm × 15 cm,4 μm). Good linearity between concentrations and peak areas was achieved in the concentration range of 5. 0-250 μmol/L for 18 kinds of amino acids. The method was validated by the analysis of five replicates. The 18 kinds of amino acid standards were spiked in fresh tea leaf samples and the average recovery rate was 86. 25%-109. 05% with relative standard deviations( n = 5) ranging from 6. 03% to 10. 56%. The limit of detection( LOD) for the analytes was0. 05-1. 27 μmol/L. The method was successfully applied to the analysis of the 18 kinds of amino acids in fresh tea leaves from east Dongting and west Dongting mountains in Suzhou. The results indicate that the method is simple,rapid,precise and reliable.展开更多
This paper reported the contents variation analysis ofγ-amino butyric acid(GABA)in Semen sojae praeparatum(SSP)which is a famous traditional Chinese medicine.High performance liquid chromatography(HPLC)was used and G...This paper reported the contents variation analysis ofγ-amino butyric acid(GABA)in Semen sojae praeparatum(SSP)which is a famous traditional Chinese medicine.High performance liquid chromatography(HPLC)was used and GABA was derivatized by online pre-column derivatization with o-phthalaldehyde(OPA).To validate this method,the precision,stability,repeatability and recovery were discussed.In the concentration range from 0.0125 to 0.400 mg/m L,the calibration curve for GABA was linear and the regression equation was obtained with correlation coefficient(R2)of 0.9999.Relatively high levels of GABA exist in SSP and the content changes of GABA at different time points during the fermenting process were detected.At the"yellow cladding"stage,GABA level was very low or even undetectable;the"secondary fermentation"stage witnessed a rapid increase of GABA content to 1.39-5.52 mg/g,which remained stable after 18 days of"secondary fermentation".This study demonstrated that GABA was generated at the"secondary fermentation"stage,revealing the significance and rationality of the"secondary fermentation"stage in the fermenting process of SSP.On the other hand,it suggested the downside of taking soy isoflavones as the only measurement in existing quality assessment and optimization approach for the fermenting process of SSP.展开更多
Sparfloxacin can be oxidized by nitrous acid, then react with hydroiodic acid to form a fluorescent derivative. Based on this, a reversed-phase high performance liquid chromatographic pre-column derivatization new met...Sparfloxacin can be oxidized by nitrous acid, then react with hydroiodic acid to form a fluorescent derivative. Based on this, a reversed-phase high performance liquid chromatographic pre-column derivatization new method is described for the determination of sparfloxacin in human urine. The linear range is 0.05 mg/L to 4.0 mg/L, the recoveries are 91.5%similar to 95.7% and the RSD is 1.2%similar to4.2%. The results showed that this method is suitable for the determination of sparfloxacin in human urine.展开更多
Gatifloxacin （GFX） is a kind of chiral fluoroquinolones compound due to the methyl group at the C-3 position of the piperazine ring[1]. Although the enantiomers of GFX show similar levels of antimicrobial activity a...Gatifloxacin （GFX） is a kind of chiral fluoroquinolones compound due to the methyl group at the C-3 position of the piperazine ring[1]. Although the enantiomers of GFX show similar levels of antimicrobial activity and pharmacokinetics[2], the other biological activities （i.e., toxicity or enantioselective recognition to various receptors in vivo） of GFX enantiomers have not yet been studied. With this in mind, we developed a rapid and cost-effective high performance liquid chromatographic （HPLC） separation procedure for GFX enantiomers with a pre-column esterification strategy.展开更多
High performance liquid chromatography method for the separation of a series of chiral benzyl alcohols on N-(3,5-dinitrobenzoyl)-D-phenylglycine stationary phase (Macherey Nagel, Chiral-2) after pre-column achiral der...High performance liquid chromatography method for the separation of a series of chiral benzyl alcohols on N-(3,5-dinitrobenzoyl)-D-phenylglycine stationary phase (Macherey Nagel, Chiral-2) after pre-column achiral derivatization was developed. Cheap and easy available aromatic acid chlorides were used as derivatization agents. Good to excellent separations of the enantiomers were achieved in all cases in relatively short analytical runs. It was shown that the enantiorecognition depends on the substituents both in the starting alcohol and in the acid chloride. The method presents an efficient alternative to the direct analyses on polysaccharide and cyclodextrine-derived stationary phases.展开更多
[Objectives]A method for the detection of monensin in poultry and livestock meat by pre-column derivatization-high performance liquid chromatography was established.[Methods]The sample was extracted with chloroform,de...[Objectives]A method for the detection of monensin in poultry and livestock meat by pre-column derivatization-high performance liquid chromatography was established.[Methods]The sample was extracted with chloroform,derivatized with trichloroacetic acid and 2,4-dinitrophenylhydrazine,and centrifuged to obtain a purified solution.A C18 chromatographic column(4.6 mm×150 mm,5μm)was used for separation with(1.5%)acetic acid water∶methanol(volume ratio)=1∶9 as the mobile phase using a DAD detector for detection,and the external standard method was adopted for peak area quantification.[Results]Monensin had good linearity in the concentration range of 5.00-200 mg/L,with the linear correlation coefficient r 2>0.999;the detection limit was 5.00 mg/kg;the relative standard deviation was smaller than 10%;and the recoveries of standard addition experiment were in the range of 75%-110%.[Conclusions]The method has the advantages of simple pretreatment operation,good derivatization effect and fast detection speed,and is suitable for detecting monensin in poultry and livestock meat.展开更多
A simple, sensitive and precise green high-performance liquid chromatographic method including on-line pre-column oxidation combined by column switching with a short Hypersil ODS analytical column (100 mm × 4.0 m...A simple, sensitive and precise green high-performance liquid chromatographic method including on-line pre-column oxidation combined by column switching with a short Hypersil ODS analytical column (100 mm × 4.0 mm i.d.) for enrichment and separation was developed and validated to determine low levels of methotrexate (MTX). The method was based on oxidative cleavage of MTX into highly fluorescence products, 2,4-diaminopteridine-6-carboxaldehyde and the corresponding 2,4-diaminopteridine-6-carboxylic acid, during the flow of phosphate buffer (0.04 M, pH 3.4) containing the analyte through the packed reactor of cerium (IV) trihydroxyhydroperoxide (CTH) at a flow-rate of 0.2 mL/min and 40℃. The fluorescent products were enriched on the head of ODS analytical column for the final separation. The separation was performed at room temperature using an environmentally friendly mobile phase consisting of ethanol and phosphate buffer (0.04 M, pH 3.4) in the ratio of 10:90 (v/v). The eluent was monitored at emission and excitation wavelengths of 463 and 367 nm, respectively. The method was successfully applied, without any interference from the excipients, for the determination of drug in tablets and vials with a detection limit of 0.06 ng/mL from 500 ?L of sample MTX.展开更多
A highly sensitive HPLC method for the detection of amino acids and oligopeptides with 9-acridine formyl chloride by pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, histidine, trigly...A highly sensitive HPLC method for the detection of amino acids and oligopeptides with 9-acridine formyl chloride by pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, histidine, triglycine and glutathione were separated on a reversed-phase C-18 column with methanol-water-triethylamine eluent, derivatization and chromatographic conditions were optimized. The five derivatives were eluted in 28 min with a good reproducibility. Linear range of the calibration graph was 0.08-260 nmol/ml(-1). The relative standard deviations(n=6) are < 5%. Detection limits (signal-to-noise ratio=3) for the five derivatives are 20-40 fmol.展开更多
Based on the chiral separation of several basic drugs, dimetindene, tetryzoline, theodrenaline and verapamil, the liquid pre-column capillary electrophoresis (LPC-CE) technique was established. It was used to determin...Based on the chiral separation of several basic drugs, dimetindene, tetryzoline, theodrenaline and verapamil, the liquid pre-column capillary electrophoresis (LPC-CE) technique was established. It was used to determine free concentrations of dmg enantiomers in mixed solutions with human serum albumin (HSA). To prevent HSA entering the CE chiral srparation zone, the mobility differences between HSA and drugs under a specific pH condition were employed in thr LPC. Thus, the detection confusion caused by protein was totally avoided. Further study of binding constants determination and protein binding competitions was carried out. The study proves that the LPC technique could be used for complex mrdia, particularly the matrix of protein coexisting with a variety of dmgs.展开更多
为建立鸽蛋中恩诺沙星、氧氟沙星、沙拉沙星和达氟沙星残留量同时检测的柱前衍生-气相色谱-串联质谱方法,以10%三氯乙酸-乙腈溶液为提取剂,提取2次,上清液经Oasis PRiME HLB柱净化后氮吹浓缩,用甲醇复溶,添加300μL三甲基硅烷基重氮甲...为建立鸽蛋中恩诺沙星、氧氟沙星、沙拉沙星和达氟沙星残留量同时检测的柱前衍生-气相色谱-串联质谱方法,以10%三氯乙酸-乙腈溶液为提取剂,提取2次,上清液经Oasis PRiME HLB柱净化后氮吹浓缩,用甲醇复溶,添加300μL三甲基硅烷基重氮甲烷衍生30 min,其衍生产物采用气相色谱-串联质谱进行确证分析。结果表明:基质匹配标准曲线在相应线性浓度范围内呈良好的线性关系,R^(2)均在0.9993以上;检测限和定量限分别为0.2~1.0和0.4~2.0μg·kg^(-1);加标回收率为78.30%~92.75%;相对标准偏差、日内相对标准偏差和日间相对标准偏差分别为2.11%~4.55%、1.04%~5.84%和3.53%~6.33%。该方法灵敏度高,定性、定量准确,符合我国兽药残留检测要求,可为禽蛋中多兽药残留监测及检测标准的制定提供技术支撑。展开更多
建立以邻苯二甲醛为衍生试剂,柱前衍生-HPLC测定特殊医学用途奶粉中牛磺酸含量的方法。选用C18色谱柱进行产物分离,并对衍生试剂用量、流动相比例、流速、混合次数、衍生后等待时间等因素进行优化。结果表明衍生试剂用量30μL,流动相体...建立以邻苯二甲醛为衍生试剂,柱前衍生-HPLC测定特殊医学用途奶粉中牛磺酸含量的方法。选用C18色谱柱进行产物分离,并对衍生试剂用量、流动相比例、流速、混合次数、衍生后等待时间等因素进行优化。结果表明衍生试剂用量30μL,流动相体积比20∶80,流速0.6 mL/min,混合6次,衍生完成后等待时间0 min时,衍生物的峰面积和峰形最优。在此实验条件下3种不同浓度牛磺酸标准品的精密度测定衍生物峰面积相对标准偏差(relative standard deviation,RSD)为0.08%~0.75%,精密度高。对两种特殊医学用途奶粉中牛磺酸含量进行检测,重复性实验测定RSD分别为0.67%和0.37%,重复性稳定;样品加标平均回收率分别为97.6%~104.5%和106.0%~110.3%,回收率好。该研究方法反应条件温和、操作性强,适用于特殊医学用途奶粉中牛磺酸含量的测定。展开更多
基金Supported by National Key Technologies R & D Program(2007BAD58B03)~~
文摘[Objective] This study aimed to establish a pre-column derivatization HPLC method for the identification and analysis of monosaccharide composition of Pu-erh tea polysaccharide. [Method] Pu-erh tea polysaccharide was extracted using the wa- ter extraction method, further isolated and purified by DEAE cellulose-52 columns. The obtained tea polysaccharide and four components TPS1, TPS2, TPS3 and TPS, were first derived by 1-phenyl-3-methyl-5-pyrazolone （PMP）, and then the PMP derivatives of monosaccharide were analyzed by high performance liquid chromatog- raphy （HPLC）. [Result] Pu-erh tea polysaccharide contained eight kinds of monosac- chaddes （mannose, rhamnose, glucuronic acid, galacturonic acid, grucose, galactose, arabinose, fucose）, without xylose; so it was the same with TPS1; each of TPS2, TPS3 and TPS4 contained seven monosaccharides, while no fucose. [Conclusion] This method is simplified and rapid, which can be used to determine the monosac- charide composition of Pu-erh tea polysaccharide and monosaccharide content.
基金Supported by Open Project of the Key Laboratory of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base(201603)Basic Research Project of Application of Suzhou City(SNG201622)
文摘A rapid and accurate quantitative method of high performance liquid chromatography( HPLC) with fluorescence detector has been developed for the analysis of 18 kinds of amino acids in fresh tea leaves. The samples were minced and mixed,and extracted with ultra pure water at 90℃ for 20 min. The 6-aminoquinolyl N-hydroxy-succinimidyl carbamate( AQC) was used as pre-column derivatization reagent. Gradient HPLC separation was performed on a C_(18) column( Symmetry C_(18),3. 9 mm × 15 cm,4 μm). Good linearity between concentrations and peak areas was achieved in the concentration range of 5. 0-250 μmol/L for 18 kinds of amino acids. The method was validated by the analysis of five replicates. The 18 kinds of amino acid standards were spiked in fresh tea leaf samples and the average recovery rate was 86. 25%-109. 05% with relative standard deviations( n = 5) ranging from 6. 03% to 10. 56%. The limit of detection( LOD) for the analytes was0. 05-1. 27 μmol/L. The method was successfully applied to the analysis of the 18 kinds of amino acids in fresh tea leaves from east Dongting and west Dongting mountains in Suzhou. The results indicate that the method is simple,rapid,precise and reliable.
基金the National Natural Science Foundation of China(81660664,82060709,82060699)the Natural Science Foundation of Jiangxi Province(20192ACBL21032,20192BBGL70051)China Scholarship Council(201908360259)。
文摘This paper reported the contents variation analysis ofγ-amino butyric acid(GABA)in Semen sojae praeparatum(SSP)which is a famous traditional Chinese medicine.High performance liquid chromatography(HPLC)was used and GABA was derivatized by online pre-column derivatization with o-phthalaldehyde(OPA).To validate this method,the precision,stability,repeatability and recovery were discussed.In the concentration range from 0.0125 to 0.400 mg/m L,the calibration curve for GABA was linear and the regression equation was obtained with correlation coefficient(R2)of 0.9999.Relatively high levels of GABA exist in SSP and the content changes of GABA at different time points during the fermenting process were detected.At the"yellow cladding"stage,GABA level was very low or even undetectable;the"secondary fermentation"stage witnessed a rapid increase of GABA content to 1.39-5.52 mg/g,which remained stable after 18 days of"secondary fermentation".This study demonstrated that GABA was generated at the"secondary fermentation"stage,revealing the significance and rationality of the"secondary fermentation"stage in the fermenting process of SSP.On the other hand,it suggested the downside of taking soy isoflavones as the only measurement in existing quality assessment and optimization approach for the fermenting process of SSP.
文摘Sparfloxacin can be oxidized by nitrous acid, then react with hydroiodic acid to form a fluorescent derivative. Based on this, a reversed-phase high performance liquid chromatographic pre-column derivatization new method is described for the determination of sparfloxacin in human urine. The linear range is 0.05 mg/L to 4.0 mg/L, the recoveries are 91.5%similar to 95.7% and the RSD is 1.2%similar to4.2%. The results showed that this method is suitable for the determination of sparfloxacin in human urine.
基金supported by Guangdong Natural Science Foundation(S2013030013338)the Ph D.Programs Foundation of Ministry of Education of China(20114404130002)Guangdong Planed Program in Science and Technology(cgzhzd0808,2013B051000072,2012A020100002)
文摘Gatifloxacin （GFX） is a kind of chiral fluoroquinolones compound due to the methyl group at the C-3 position of the piperazine ring[1]. Although the enantiomers of GFX show similar levels of antimicrobial activity and pharmacokinetics[2], the other biological activities （i.e., toxicity or enantioselective recognition to various receptors in vivo） of GFX enantiomers have not yet been studied. With this in mind, we developed a rapid and cost-effective high performance liquid chromatographic （HPLC） separation procedure for GFX enantiomers with a pre-column esterification strategy.
文摘High performance liquid chromatography method for the separation of a series of chiral benzyl alcohols on N-(3,5-dinitrobenzoyl)-D-phenylglycine stationary phase (Macherey Nagel, Chiral-2) after pre-column achiral derivatization was developed. Cheap and easy available aromatic acid chlorides were used as derivatization agents. Good to excellent separations of the enantiomers were achieved in all cases in relatively short analytical runs. It was shown that the enantiorecognition depends on the substituents both in the starting alcohol and in the acid chloride. The method presents an efficient alternative to the direct analyses on polysaccharide and cyclodextrine-derived stationary phases.
文摘[Objectives]A method for the detection of monensin in poultry and livestock meat by pre-column derivatization-high performance liquid chromatography was established.[Methods]The sample was extracted with chloroform,derivatized with trichloroacetic acid and 2,4-dinitrophenylhydrazine,and centrifuged to obtain a purified solution.A C18 chromatographic column(4.6 mm×150 mm,5μm)was used for separation with(1.5%)acetic acid water∶methanol(volume ratio)=1∶9 as the mobile phase using a DAD detector for detection,and the external standard method was adopted for peak area quantification.[Results]Monensin had good linearity in the concentration range of 5.00-200 mg/L,with the linear correlation coefficient r 2>0.999;the detection limit was 5.00 mg/kg;the relative standard deviation was smaller than 10%;and the recoveries of standard addition experiment were in the range of 75%-110%.[Conclusions]The method has the advantages of simple pretreatment operation,good derivatization effect and fast detection speed,and is suitable for detecting monensin in poultry and livestock meat.
文摘A simple, sensitive and precise green high-performance liquid chromatographic method including on-line pre-column oxidation combined by column switching with a short Hypersil ODS analytical column (100 mm × 4.0 mm i.d.) for enrichment and separation was developed and validated to determine low levels of methotrexate (MTX). The method was based on oxidative cleavage of MTX into highly fluorescence products, 2,4-diaminopteridine-6-carboxaldehyde and the corresponding 2,4-diaminopteridine-6-carboxylic acid, during the flow of phosphate buffer (0.04 M, pH 3.4) containing the analyte through the packed reactor of cerium (IV) trihydroxyhydroperoxide (CTH) at a flow-rate of 0.2 mL/min and 40℃. The fluorescent products were enriched on the head of ODS analytical column for the final separation. The separation was performed at room temperature using an environmentally friendly mobile phase consisting of ethanol and phosphate buffer (0.04 M, pH 3.4) in the ratio of 10:90 (v/v). The eluent was monitored at emission and excitation wavelengths of 463 and 367 nm, respectively. The method was successfully applied, without any interference from the excipients, for the determination of drug in tablets and vials with a detection limit of 0.06 ng/mL from 500 ?L of sample MTX.
文摘A highly sensitive HPLC method for the detection of amino acids and oligopeptides with 9-acridine formyl chloride by pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, histidine, triglycine and glutathione were separated on a reversed-phase C-18 column with methanol-water-triethylamine eluent, derivatization and chromatographic conditions were optimized. The five derivatives were eluted in 28 min with a good reproducibility. Linear range of the calibration graph was 0.08-260 nmol/ml(-1). The relative standard deviations(n=6) are < 5%. Detection limits (signal-to-noise ratio=3) for the five derivatives are 20-40 fmol.
文摘Based on the chiral separation of several basic drugs, dimetindene, tetryzoline, theodrenaline and verapamil, the liquid pre-column capillary electrophoresis (LPC-CE) technique was established. It was used to determine free concentrations of dmg enantiomers in mixed solutions with human serum albumin (HSA). To prevent HSA entering the CE chiral srparation zone, the mobility differences between HSA and drugs under a specific pH condition were employed in thr LPC. Thus, the detection confusion caused by protein was totally avoided. Further study of binding constants determination and protein binding competitions was carried out. The study proves that the LPC technique could be used for complex mrdia, particularly the matrix of protein coexisting with a variety of dmgs.
文摘为建立鸽蛋中恩诺沙星、氧氟沙星、沙拉沙星和达氟沙星残留量同时检测的柱前衍生-气相色谱-串联质谱方法,以10%三氯乙酸-乙腈溶液为提取剂,提取2次,上清液经Oasis PRiME HLB柱净化后氮吹浓缩,用甲醇复溶,添加300μL三甲基硅烷基重氮甲烷衍生30 min,其衍生产物采用气相色谱-串联质谱进行确证分析。结果表明:基质匹配标准曲线在相应线性浓度范围内呈良好的线性关系,R^(2)均在0.9993以上;检测限和定量限分别为0.2~1.0和0.4~2.0μg·kg^(-1);加标回收率为78.30%~92.75%;相对标准偏差、日内相对标准偏差和日间相对标准偏差分别为2.11%~4.55%、1.04%~5.84%和3.53%~6.33%。该方法灵敏度高,定性、定量准确,符合我国兽药残留检测要求,可为禽蛋中多兽药残留监测及检测标准的制定提供技术支撑。
文摘建立以邻苯二甲醛为衍生试剂,柱前衍生-HPLC测定特殊医学用途奶粉中牛磺酸含量的方法。选用C18色谱柱进行产物分离,并对衍生试剂用量、流动相比例、流速、混合次数、衍生后等待时间等因素进行优化。结果表明衍生试剂用量30μL,流动相体积比20∶80,流速0.6 mL/min,混合6次,衍生完成后等待时间0 min时,衍生物的峰面积和峰形最优。在此实验条件下3种不同浓度牛磺酸标准品的精密度测定衍生物峰面积相对标准偏差(relative standard deviation,RSD)为0.08%~0.75%,精密度高。对两种特殊医学用途奶粉中牛磺酸含量进行检测,重复性实验测定RSD分别为0.67%和0.37%,重复性稳定;样品加标平均回收率分别为97.6%~104.5%和106.0%~110.3%,回收率好。该研究方法反应条件温和、操作性强,适用于特殊医学用途奶粉中牛磺酸含量的测定。
