[Objectives]To establish an efficient and environmentally friendly separation and purification method for the large-scale preparation of the major active components-eleutherol,eleutherine,and isoeleutherine-from the e...[Objectives]To establish an efficient and environmentally friendly separation and purification method for the large-scale preparation of the major active components-eleutherol,eleutherine,and isoeleutherine-from the ethnomedicinal plant Eleutherine americana Merr.et K.Heyne.[Methods]The sample of E.americana bulbs was initially extracted with ethanol,followed by three successive extractions with ethyl acetate-water(2:1,V/V)to obtain the target component-enriched fraction.Eight solvent systems were systematically optimized,and a mixture of petroleum ether-ethyl acetate-ethanol-water(5:5:6:4,V/V/V/V)was identified as the optimal solvent system for high-speed counter-current chromatography(HSCCC)separation under conditions of 900 rpm,2 mL/min,and 35℃.The crude HSCCC product was further purified by silica gel column chromatography(200-300 mesh)using gradient elution with a solvent system of n-hexane-dichloromethane-ethyl acetate(varying from 10:5:1 to 4:5:1,V/V/V).UPLC-PDA(Agilent SB-C_(18)column)and nuclear magnetic resonance spectroscopy(600 MHz)were comprehensively employed to assess compound purity and confirm molecular structures.[Results]An optimized technique integrating HSCCC and silica gel column chromatography was established,successfully enabling the large-scale preparation of three bioactive components:eleutherol(purity 99%),eleutherine(purity 98%),and isoeleutherine(purity 98%).Structural identification results were consistent with those reported in the literature.Compared to traditional methods,the new approach demonstrated improved separation efficiency and reduced solvent consumption.[Conclusions]The combined separation method utilizing HSCCC and silica gel column chromatography established in this study demonstrates notable advantages,including high efficiency,environmental friendliness,and cost-effectiveness,enabling the large-scale preparation of the three major active components from E.americana.This approach outperforms conventional methods by offering higher separation efficiency,reduced solvent consumption,and superior product purity,providing a robust technical solution for the development and utilization of bioactive compounds from E.americana.Moreover,it offers a novel methodological reference for the isolation and purification of other natural products.展开更多
Quantitative structure-retention relationship(QSRR)is an important tool in chromatography.QSRR examines the correlation between molecular structures and their retention behaviors during chromatographic separation.This...Quantitative structure-retention relationship(QSRR)is an important tool in chromatography.QSRR examines the correlation between molecular structures and their retention behaviors during chromatographic separation.This approach involves developing models for predicting the retention time(RT)of analytes,thereby accelerating method development and facilitating compound identification.In addition,QSRR can be used to study compound retention mechanisms and support drug screening efforts.This review provides a comprehensive analysis of QSRR workflows and applications,with a special focus on the role of artificial intelligence-an area not thoroughly explored in previous reviews.Moreover,we discuss current limitations in RT prediction and propose promising solutions.Overall,this review offers a fresh perspective on future QSRR research,encouraging the development of innovative strategies that enable the diverse applications of QSRR models in chromatographic analysis.展开更多
[Objectives]To explore the methods for identifying pure honey.[Methods]Using 35 batches of Jiulongteng honey sourced from various production areas in Guangxi as the research subjects,this study investigated the chromo...[Objectives]To explore the methods for identifying pure honey.[Methods]Using 35 batches of Jiulongteng honey sourced from various production areas in Guangxi as the research subjects,this study investigated the chromogenic reactions of starch and dextrin,as well as the comparative study of thin-layer chromatography of oligosaccharides present in Jiulongteng honey.[Results]None of the 35 batches of Jiulongteng honey samples exhibited blue(indicating starch),green,or reddish-brown(indicating dextrin)coloration,suggesting that no adulterants such as artificially added starch,dextrin,or sugar were present in these samples.Furthermore,none of the 35 batches displayed additional spots below the corresponding positions of the control,indicating that the sugar composition was consistent with the oligosaccharide profile of natural honey.No components inconsistent with the oligosaccharide profile of natural honey were detected.Therefore,it can be concluded that the Jiulongteng honey samples in this experiment were pure and free from adulteration with starch,dextrin,or other sugar substances.[Conclusions]The method employed in this experiment is straightforward and quick to implement,effectively preventing adulterated honey from entering the market.It enhances the efficiency of quality control for Jiulongteng honey and promotes the healthy development of the Jiulongteng honey industry.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
A high-performance liquid chromatography(HPLC)method has been developed using a CAPCELL PAK ADME(150 mm×4.6 mm,5μm)as analytical column and a gradient elution with 15 min using acetonitrile and 0.1%(in volume fr...A high-performance liquid chromatography(HPLC)method has been developed using a CAPCELL PAK ADME(150 mm×4.6 mm,5μm)as analytical column and a gradient elution with 15 min using acetonitrile and 0.1%(in volume fraction)phosphoric acid water(pH=2.2)as the mobile phase.Three active substances in cosmetics were quantitatively detected simultaneously at a detection wavelength of 265 nm.The linear ranges of β-nicotinamide mononucleotides,ergothioneine and nicotinamide are 10~200 mg/L,5~100 mg/L and 5~100 mg/L respectively and the detection limits of three components are 3.0 mg/L,1.5 mg/L and 1.5 mg/L respectively.The recovery rate is 97.1~104.9%,with RSD≤2.0%.The method was applied to quantitative analysis of five samples of cosmetics toner,lotion,cream,essence and gel and three samples of raw materials.The results showed that the results of β-nicotinamide mononucleotide and ergothioneine in five cosmetics were consistent with the product label,while nicotinamide was inconsistent with the label.The purity of the three raw material samples was 99.5%,99.7% and 100.8%respectively.This method offers high precision,accuracy and short analysis time,making it a reliable approach for studying three active ingredients in cosmetics and suitable for quality control of related functional ingredients.展开更多
[Objective]The aim of this work was to establish an analytical method for the determination of deoxynivalenol(DON)in feeds using automatic immunomagnetic beads(IMBs)clean-up coupled with high-performance liquid chroma...[Objective]The aim of this work was to establish an analytical method for the determination of deoxynivalenol(DON)in feeds using automatic immunomagnetic beads(IMBs)clean-up coupled with high-performance liquid chromatography.[Method]Feed samples were extracted using ultra-pure water,purified by automatic IMBs,and subsequently analyzed via high-performance liquid chromatography,employing an external standard method for quantification.[Result]A satisfactory linearity was achieved for DON within the concentration range of 0.05 to 2.0μg/mL,with the corresponding correlation coefficients(R^(2))exceeding 0.9999.The limit of detection(LOD)and limit of quantification(LOQ)for the proposed method were determined to be 0.03 and 0.1 mg/kg,respectively.The average recoveries of the fortified samples(0.1,0.2 and 1.0 mg/kg)were 88.5%−100.6%,with the relative standard deviations(RSD)ranging from 2.1%to 9.7%.[Conclusion]In comparison with the traditional solidphase extraction and immunoaffinity column purification methods,the IMBs technique consolidates the extraction,separation,and purification into a single process.This approach enables fully automated processing,which significantly enhances work efficiency and mitigates result deviations that may arise from manual operations.Consequently,this technique is particularly well-suited for the determination of DON in a large number of feed samples.展开更多
Background:Polygonum multiflorum-induced liver injury(PM-DILI)has significantly hindered its clinical application and development.Methods:This study investigates the variation in content and toxicity of dian-thrones,t...Background:Polygonum multiflorum-induced liver injury(PM-DILI)has significantly hindered its clinical application and development.Methods:This study investigates the variation in content and toxicity of dian-thrones,the toxic components of P.multiflorum,during different processing cycles.We employed the ultra-high-performance liquid chromatography triple quadrupole mass spectrometry method to quantify six dianthrones in raw P.multiflorum and formulations processed with a method called nine cycles of steaming and sunning.Additionally,toxicity assessments were conducted using human normal liver cell line L02 and zebrafish embryos.Results:Results indicate a gradual reduction in dianthrones content with increasing processing cycles.Processed formulations exhibited significantly reduced cytotoxic-ity in L02 cells and hepatotoxicity in zebrafish embryos.Conclusions:Our findings elucidate the relationship between processing cycles and P.multiflorum toxicity,providing theoretical support for its safe use.展开更多
Arsenic speciation in freshwater fish is crucial for providing meaningful consumption guidelines that allow the public to make informed decisions regarding its consumption.While marine fish have attractedmuch research...Arsenic speciation in freshwater fish is crucial for providing meaningful consumption guidelines that allow the public to make informed decisions regarding its consumption.While marine fish have attractedmuch research interest due to their higher arsenic content,research on freshwater fish is limited due to the challenges in quantifying and identifying arsenic species present at trace levels.We describe here a sensitivemethod and its application to the quantification of arsenic species in freshwater fish.Arsenic species from fish tissues were extracted using a methanol/water mixture(1:1 vol.ratio)and ultrasound sonication.Anion-exchange high-performance liquid chromatography(HPLC)enabled separation of arsenobetaine(AsB),inorganic arsenite(iAs^(Ⅲ)),dimethylarsinic acid(DMA),monomethylarsonic acid(MMA),inorganic arsenate(iAs^(Ⅴ)),and three new arsenic species.Inductively coupled plasma mass spectrometry(ICPMS)provided highly sensitive and specific detection of arsenic.A limit of detection of 0.25μg/kg(wet weight fish tissue)was achieved for the five target arsenic species:AsB,iAs^(Ⅲ),DMA,MMA,and iAs^(Ⅴ).A series of experimentswere conducted to ensure the accuracy and validity of the analytical method.The method was successfully applied to the determination of arsenic species in lakewhitefish,northern pike,and walleye,with AsB,DMA,and iAs^(Ⅴ) being frequently detected.Three new arsenic species were detected,but their chromatographic retention times did not match with those of any available arsenic standards.Future research is necessary to elucidate the identity of these new arsenic species detected in freshwater fish.展开更多
Aim To establish a reversed-phase liquid chromatographic (LC) method forsimultaneous determination of tetramethylpyrazine (TMP) and aspirin in a new compound formulation.Methods Chromatographic separation of the two d...Aim To establish a reversed-phase liquid chromatographic (LC) method forsimultaneous determination of tetramethylpyrazine (TMP) and aspirin in a new compound formulation.Methods Chromatographic separation of the two drugs was achieved on a Diamonsil C_(18) column, usinga binary mixture of methanol-1.5% acetic acid (35:65, V/V, pH = 3.1) as mobile phase at a flow rateof 1.0 mL·min^(-1). Results Separation was completed in less than 12 min. Benzoic acid was used asthe internal standard. Recoveries at levels corresponding to 80 % to 120 % of the label claim ofthe formulation ranged from 99.6 to 100.3 % for aspirin and from 99.9 to 101.3% for TMP. The linearrange was 12.6 - 150.9 μg·mL^(-1)(r= 0.9997, n = 5) for aspirin and 25.0- 300.0 μg·mL^(-1) (r =0.9999, n = 5) for TMP. Conclusion The method developed can be used for the simultaneousdetermination of TMP and aspirin in pharmaceutical preparations.展开更多
Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on dete...Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on determination of residual protein in lincomycin hydrochloride. Methods The chromatographic conditions were SuperdexTM peptide column, 0.01 mol*L-1 phosphate buffer solution as mobile phase, and flow rate of 1 mL·min-1. Five hundred microliters of lincomycin hydrochloride solution (3 g of lincomycin hydrochloride dissolved in 10 mL of mobile phase) was injected into the chromatograph and the eluted solution was collected between 6 min and 14.5 min (protein eluted from column within this period), and the residual content of total protein in the eluted solution was assayed using Bradford assay method. Results The average recovery was more than 90% for bovine serum albumin, the calibration equation for the range of 0-12 μg·mL-1 of protein was y=-0.002 4x2+0.064 2x+0.002 9, r2=0.999 9, RSD=0.1%-0.9%, and the LOD and LOQ were 3 and 10 ng·mL-1 of protein, respectively. Conclusion The novel method for determining the residual protein in ferment antibio-tics is simple, rapid, and precise.展开更多
基金Supported by Natural Science Foundation of Tibet Autonomous Region,Science and Technology Department of Tibet(XZ202501ZR0118).
文摘[Objectives]To establish an efficient and environmentally friendly separation and purification method for the large-scale preparation of the major active components-eleutherol,eleutherine,and isoeleutherine-from the ethnomedicinal plant Eleutherine americana Merr.et K.Heyne.[Methods]The sample of E.americana bulbs was initially extracted with ethanol,followed by three successive extractions with ethyl acetate-water(2:1,V/V)to obtain the target component-enriched fraction.Eight solvent systems were systematically optimized,and a mixture of petroleum ether-ethyl acetate-ethanol-water(5:5:6:4,V/V/V/V)was identified as the optimal solvent system for high-speed counter-current chromatography(HSCCC)separation under conditions of 900 rpm,2 mL/min,and 35℃.The crude HSCCC product was further purified by silica gel column chromatography(200-300 mesh)using gradient elution with a solvent system of n-hexane-dichloromethane-ethyl acetate(varying from 10:5:1 to 4:5:1,V/V/V).UPLC-PDA(Agilent SB-C_(18)column)and nuclear magnetic resonance spectroscopy(600 MHz)were comprehensively employed to assess compound purity and confirm molecular structures.[Results]An optimized technique integrating HSCCC and silica gel column chromatography was established,successfully enabling the large-scale preparation of three bioactive components:eleutherol(purity 99%),eleutherine(purity 98%),and isoeleutherine(purity 98%).Structural identification results were consistent with those reported in the literature.Compared to traditional methods,the new approach demonstrated improved separation efficiency and reduced solvent consumption.[Conclusions]The combined separation method utilizing HSCCC and silica gel column chromatography established in this study demonstrates notable advantages,including high efficiency,environmental friendliness,and cost-effectiveness,enabling the large-scale preparation of the three major active components from E.americana.This approach outperforms conventional methods by offering higher separation efficiency,reduced solvent consumption,and superior product purity,providing a robust technical solution for the development and utilization of bioactive compounds from E.americana.Moreover,it offers a novel methodological reference for the isolation and purification of other natural products.
基金supported by the Shanghai Sailing Program,China(Grant No.:23YF1413300).
文摘Quantitative structure-retention relationship(QSRR)is an important tool in chromatography.QSRR examines the correlation between molecular structures and their retention behaviors during chromatographic separation.This approach involves developing models for predicting the retention time(RT)of analytes,thereby accelerating method development and facilitating compound identification.In addition,QSRR can be used to study compound retention mechanisms and support drug screening efforts.This review provides a comprehensive analysis of QSRR workflows and applications,with a special focus on the role of artificial intelligence-an area not thoroughly explored in previous reviews.Moreover,we discuss current limitations in RT prediction and propose promising solutions.Overall,this review offers a fresh perspective on future QSRR research,encouraging the development of innovative strategies that enable the diverse applications of QSRR models in chromatographic analysis.
基金Supported by Enhancement Project of Basic Scientific Research Ability of Young and Middle-aged Teachers in Guangxi Universities(2020KY07040)School-level Scientific Research Project of Guangxi University of Chinese Medicine(2024QN022)Self-financed Scientific Research Project of Guangxi Zhuang Autonomous Region Administration of Traditional Chinese Medicine(GXZYL20240818).
文摘[Objectives]To explore the methods for identifying pure honey.[Methods]Using 35 batches of Jiulongteng honey sourced from various production areas in Guangxi as the research subjects,this study investigated the chromogenic reactions of starch and dextrin,as well as the comparative study of thin-layer chromatography of oligosaccharides present in Jiulongteng honey.[Results]None of the 35 batches of Jiulongteng honey samples exhibited blue(indicating starch),green,or reddish-brown(indicating dextrin)coloration,suggesting that no adulterants such as artificially added starch,dextrin,or sugar were present in these samples.Furthermore,none of the 35 batches displayed additional spots below the corresponding positions of the control,indicating that the sugar composition was consistent with the oligosaccharide profile of natural honey.No components inconsistent with the oligosaccharide profile of natural honey were detected.Therefore,it can be concluded that the Jiulongteng honey samples in this experiment were pure and free from adulteration with starch,dextrin,or other sugar substances.[Conclusions]The method employed in this experiment is straightforward and quick to implement,effectively preventing adulterated honey from entering the market.It enhances the efficiency of quality control for Jiulongteng honey and promotes the healthy development of the Jiulongteng honey industry.
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.
文摘A high-performance liquid chromatography(HPLC)method has been developed using a CAPCELL PAK ADME(150 mm×4.6 mm,5μm)as analytical column and a gradient elution with 15 min using acetonitrile and 0.1%(in volume fraction)phosphoric acid water(pH=2.2)as the mobile phase.Three active substances in cosmetics were quantitatively detected simultaneously at a detection wavelength of 265 nm.The linear ranges of β-nicotinamide mononucleotides,ergothioneine and nicotinamide are 10~200 mg/L,5~100 mg/L and 5~100 mg/L respectively and the detection limits of three components are 3.0 mg/L,1.5 mg/L and 1.5 mg/L respectively.The recovery rate is 97.1~104.9%,with RSD≤2.0%.The method was applied to quantitative analysis of five samples of cosmetics toner,lotion,cream,essence and gel and three samples of raw materials.The results showed that the results of β-nicotinamide mononucleotide and ergothioneine in five cosmetics were consistent with the product label,while nicotinamide was inconsistent with the label.The purity of the three raw material samples was 99.5%,99.7% and 100.8%respectively.This method offers high precision,accuracy and short analysis time,making it a reliable approach for studying three active ingredients in cosmetics and suitable for quality control of related functional ingredients.
基金Supported by National Key Research and Development Program of China(2023YFD1301001)Central Public-interest Scientific Institution Basal Research Fund(1610072023005)Agricultural Science and Technology Innovation Program of CAAS(CAAS-ASTIP-IQSTAP-04).
文摘[Objective]The aim of this work was to establish an analytical method for the determination of deoxynivalenol(DON)in feeds using automatic immunomagnetic beads(IMBs)clean-up coupled with high-performance liquid chromatography.[Method]Feed samples were extracted using ultra-pure water,purified by automatic IMBs,and subsequently analyzed via high-performance liquid chromatography,employing an external standard method for quantification.[Result]A satisfactory linearity was achieved for DON within the concentration range of 0.05 to 2.0μg/mL,with the corresponding correlation coefficients(R^(2))exceeding 0.9999.The limit of detection(LOD)and limit of quantification(LOQ)for the proposed method were determined to be 0.03 and 0.1 mg/kg,respectively.The average recoveries of the fortified samples(0.1,0.2 and 1.0 mg/kg)were 88.5%−100.6%,with the relative standard deviations(RSD)ranging from 2.1%to 9.7%.[Conclusion]In comparison with the traditional solidphase extraction and immunoaffinity column purification methods,the IMBs technique consolidates the extraction,separation,and purification into a single process.This approach enables fully automated processing,which significantly enhances work efficiency and mitigates result deviations that may arise from manual operations.Consequently,this technique is particularly well-suited for the determination of DON in a large number of feed samples.
基金National Natural Science Foundation of China,Grant/Award Number:81973476Chinese Society of Toxicology,Grant/Award Number:CST2021CT101。
文摘Background:Polygonum multiflorum-induced liver injury(PM-DILI)has significantly hindered its clinical application and development.Methods:This study investigates the variation in content and toxicity of dian-thrones,the toxic components of P.multiflorum,during different processing cycles.We employed the ultra-high-performance liquid chromatography triple quadrupole mass spectrometry method to quantify six dianthrones in raw P.multiflorum and formulations processed with a method called nine cycles of steaming and sunning.Additionally,toxicity assessments were conducted using human normal liver cell line L02 and zebrafish embryos.Results:Results indicate a gradual reduction in dianthrones content with increasing processing cycles.Processed formulations exhibited significantly reduced cytotoxic-ity in L02 cells and hepatotoxicity in zebrafish embryos.Conclusions:Our findings elucidate the relationship between processing cycles and P.multiflorum toxicity,providing theoretical support for its safe use.
基金supported by Alberta Health,Alberta Innovates,the Canada Research Chairs Program,the Canadian Institutes of Health Research,and the Natural Sciences and Engineering Research Council of Canada。
文摘Arsenic speciation in freshwater fish is crucial for providing meaningful consumption guidelines that allow the public to make informed decisions regarding its consumption.While marine fish have attractedmuch research interest due to their higher arsenic content,research on freshwater fish is limited due to the challenges in quantifying and identifying arsenic species present at trace levels.We describe here a sensitivemethod and its application to the quantification of arsenic species in freshwater fish.Arsenic species from fish tissues were extracted using a methanol/water mixture(1:1 vol.ratio)and ultrasound sonication.Anion-exchange high-performance liquid chromatography(HPLC)enabled separation of arsenobetaine(AsB),inorganic arsenite(iAs^(Ⅲ)),dimethylarsinic acid(DMA),monomethylarsonic acid(MMA),inorganic arsenate(iAs^(Ⅴ)),and three new arsenic species.Inductively coupled plasma mass spectrometry(ICPMS)provided highly sensitive and specific detection of arsenic.A limit of detection of 0.25μg/kg(wet weight fish tissue)was achieved for the five target arsenic species:AsB,iAs^(Ⅲ),DMA,MMA,and iAs^(Ⅴ).A series of experimentswere conducted to ensure the accuracy and validity of the analytical method.The method was successfully applied to the determination of arsenic species in lakewhitefish,northern pike,and walleye,with AsB,DMA,and iAs^(Ⅴ) being frequently detected.Three new arsenic species were detected,but their chromatographic retention times did not match with those of any available arsenic standards.Future research is necessary to elucidate the identity of these new arsenic species detected in freshwater fish.
文摘Aim To establish a reversed-phase liquid chromatographic (LC) method forsimultaneous determination of tetramethylpyrazine (TMP) and aspirin in a new compound formulation.Methods Chromatographic separation of the two drugs was achieved on a Diamonsil C_(18) column, usinga binary mixture of methanol-1.5% acetic acid (35:65, V/V, pH = 3.1) as mobile phase at a flow rateof 1.0 mL·min^(-1). Results Separation was completed in less than 12 min. Benzoic acid was used asthe internal standard. Recoveries at levels corresponding to 80 % to 120 % of the label claim ofthe formulation ranged from 99.6 to 100.3 % for aspirin and from 99.9 to 101.3% for TMP. The linearrange was 12.6 - 150.9 μg·mL^(-1)(r= 0.9997, n = 5) for aspirin and 25.0- 300.0 μg·mL^(-1) (r =0.9999, n = 5) for TMP. Conclusion The method developed can be used for the simultaneousdetermination of TMP and aspirin in pharmaceutical preparations.
文摘Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on determination of residual protein in lincomycin hydrochloride. Methods The chromatographic conditions were SuperdexTM peptide column, 0.01 mol*L-1 phosphate buffer solution as mobile phase, and flow rate of 1 mL·min-1. Five hundred microliters of lincomycin hydrochloride solution (3 g of lincomycin hydrochloride dissolved in 10 mL of mobile phase) was injected into the chromatograph and the eluted solution was collected between 6 min and 14.5 min (protein eluted from column within this period), and the residual content of total protein in the eluted solution was assayed using Bradford assay method. Results The average recovery was more than 90% for bovine serum albumin, the calibration equation for the range of 0-12 μg·mL-1 of protein was y=-0.002 4x2+0.064 2x+0.002 9, r2=0.999 9, RSD=0.1%-0.9%, and the LOD and LOQ were 3 and 10 ng·mL-1 of protein, respectively. Conclusion The novel method for determining the residual protein in ferment antibio-tics is simple, rapid, and precise.
