液相分离及联用技术在药物分析和化妆品中限用禁用物质分析中的应用研究
摘要
液相分离方法中最为成熟和有效的方法当属高效液相色谱和毛细管电泳。高效液相色谱法(High Performance Liqui Chromatography,HPLC)作为一种发展较为成熟的分析技术,以其分离效率,稳定性,重复性等特点在药物分析包括化妆品中限用禁用药物的分析方面发挥着重要的作用。而毛细管电泳(CapillaryElectrophoresis,CE)是近二十年来发展较快的分析分离技术之一,具有分离效率高、分析速度快、所需样品少等特点,应用范围广。在绪论中概述了两种分析技术发展的历史、现状以及发展趋势,对两种分析技术的基本原理和特点进行了简单介绍,并介绍了近年来两种分析检测技术在药物分析、药品质量控制、化妆品中限用禁用药物安全性控制等方面的应用。
本论文采用高效液相色谱法(HPLC)-电化学检测(ECD)联用技术对药物的杂质进行了分离测定,比较了HPLC-ECD和HPLC-UV的区别。分别采用快速液相色谱法(RRLC)-光电二极管阵列检测器(DAD)联用技术,高效毛细管电泳(HPCE)-电化学检测(ECD)联用技术,高效液相色谱法(HPLC)-串联质谱检测(MS/MS)联用技术测定了化妆品中的限用禁用药物,主要内容如下:
1.甲氨蝶呤注射液含量及有关物质的HPLC-ECD法分离测定研究
以乙腈-0.2mol·L~(-1)磷酸盐缓冲液(pH6.0)(10∶90)为流动相,采用TSK-GELODS-100SP色谱柱(150mm×4.6mm,5μm),流速为1.0mL·min~(-1),柱温35℃时,在+0.8V电位处,以玻碳圆盘电极为工作电极,银-氯化银电极为参比电极,建立了以高效液相色谱电化学检测对甲氨蝶呤及其有关物质进行分离分析的方法。甲氨蝶呤的线性范围为9.1-91μg·mL~(-1),r=0.9999;平均回收率为100.4%,RSD为1.0%(n=9)。氨蝶呤、甲蝶呤和甲氨蝶呤的最低检出量分别为3.3、11和8.6ng。电化学安培检测与紫外检测相比,具有较高的灵敏度,能有效地应用于甲氨蝶呤注射液中甲氨蝶呤含量以及极微量有关物质的检测。
2.替尼泊苷注射液含量及有关物质的HPLC-ECD法分离检测研究
建立以高效液相色谱电化学检测对替尼泊苷及其有关物质进行分离分析的方法。以PhenomenexR LUNA Phenyl-Hexyl柱,以乙腈-水(38∶62)为流动相,流速为1.0mL.min~(-1),玻碳圆盘电极为工作电极,银-氯化银电极为参比电极,在+0.7V电位处,实现了替尼泊苷及其有关物质与注射液中各种赋形剂的分离分析。木酚素、替尼泊苷、α-亚乙基木酚素P和苦亚乙基木酚素P分别在0.1-9.6μg.mL~(-1),0.1-9.0μg.mL~(-1),0.1-5.8μg.mL~(-1)和0.1-7.2μg.mL~(-1)浓度范围内呈良好线性,检测限(LOD,S/N=3)分别为0.2ng,1.1ng,1.0ng和0.7ng。电化学安培检测与紫外检测相比,具有较高的灵敏度和选择性,能有效地应用于替尼泊苷注射液中有关物质的检测和含量的测定。
3.恩他卡朋含量及有关物质的HPLC-ECD法分离测定研究
建立以高效液相色谱-电化学检测对恩他卡朋及其有关物质(z-异构体和3,4-二羟基-5-硝基苯甲醛)进行分离分析的方法。采用苯基柱,以甲醇-磷酸盐缓冲溶液(pH 2.1)(46∶54)为流动相,玻碳圆盘电极为工作电极,银-氯化银电极为参比电极,在+0.65V工作电位处,实现了恩他卡朋及其有关物质的分离分析。恩他卡朋在0.06～38.6μg mL~(-1)、z-异构体在0.06～42.5μg mL~(-1)、3,4-二羟基-5-硝基苯甲醛在0.06～27.5μg mL~(-1)浓度范围内线性关系良好,检测限分别为0.05、0.06和0.03ng。恩他卡朋的平均回收率为99.9%,RSD为1.5%。电化学安培检测具有较高的灵敏度和选择性,能有效地应用于恩他卡朋原料和片剂中有关物质的检测和片剂含量的测定。
4.胶束毛细管电泳安培检测法分离测定化妆品中限用禁用酚类化合物
采用毛细管电泳-电化学检测技术(CE-ECD)同时测定了化妆品中限用禁用酚类化合物。考察了工作电极的氧化电位、运行缓冲液的酸度和浓度、分离电压、进样时间等因素对分离和检测的影响。在优化条件下,以直径300μm的碳圆盘电极为工作电极,工作电位为+0.80 V(vs.SCE),在40 mmol L~(-1)pH9.0的H_3BO_3-Na_2B_4O_7缓冲液(含10 mmol L~(-1) SDS)中,上述组分实现了较好分离。被测物浓度与峰电流在三个数量级范围内呈良好线性关系,检测限在4.09×10~(-8)g.mL~(-1)到4.50×10~(-7)g.mL~(-1)之间,该方法简单可靠,已成功应用于不同化妆品中限用禁用酚类化合物的测定,是一种有效的化妆品安全分析检测方法。
5.RPLC-DAD法分离测定化妆品中抗生素
建立了祛痘除螨类化妆品中8种抗生素及甲硝唑同时测定的快速高效液相色谱分析方法。以Agielent1200快速高效液相色谱,采用SB和XDB-C18(1.8μm,4.6mm i.d.×50 mm)色谱柱,0.1 mol L~(-1)磷酸二氢钾溶液(用磷酸调节pH为2.5)-乙腈为流动相进行梯度洗脱,在7分钟内首次实现了8种抗生素及甲硝唑的同时分离测定,常规液相色谱分离至少需要30分钟才能完成一次分析。在4～100μg.mL~(-1)线性范围内,9种物质的线性相关系数均＞0.9999,最低检出限为0.25～1.4ng。在低中高三个添加水平,9种物质的平均回收率为92.2%～103.2%,相对标准偏差为0.04%～4.5%。并考察了不同状态的化妆品样品的提取方式。该法操作简便、准确、快速,能够检测祛痘除螨类化妆品中8种抗生素以及甲硝唑的含量。
6.HPLC-MS/MS测定化妆品中丙烯酰胺残留单体
采用高效液相色谱-串联质谱,Atlantis C18色谱柱(2.1×150 mm,3μm),以0.1%甲酸溶液-0.1%甲酸甲醇溶液(98∶2)为流动相A,0.1%甲酸甲醇溶液为流动相B进行梯度洗脱。添加同位素内标~(13)C3-丙烯酰胺于样品中,经0.1%甲酸溶液提取,石油醚液液萃取净化,串联质谱多反应监测模式,以保留时间及质谱图定性,以峰高或峰面积进行定量,测定化妆品中丙烯酰胺单体残留量。采用本方法的检出限为0.01 ng,最低检出浓度为0.01mg/Kg。方法适用于化妆品中丙烯酰胺单体残留量的测定,方法灵敏度高、重现性好。
High performance liquid chromatography(HPLC)and high performance capillary electrophoresis(HPCE)are the most important separation techniques in liquid separation method.The principle,character,and key factors of HPLC and HPCE technologies were described.The history,current situation and future trends of HPLC and HPCE were reviewed in this thesis.HPLC has greatly evolved into a major analytical technique that is routinely employed for the rapid,reliable and highly sensitive determination of numerous compounds in pharmaceutical analysis and quality control of cosmetic products.While HPCE also plays an important role in determination of impurities of pharmaceutical analysis,in chiral separation,as well as in illegal adulteration in cosmetic products,due to its speed,efficiency,reproducibility, ultra-small sample volume and ease of clearing up the contaminants.This review covers around 269 references including all types of chromatographic and hyphenated techniques reported in the latest two years.
The present research work describes the studies of high performance liquid chromatography with electrochemical detection in the quality control of impurities in pharmaceutical preparations,and different types of chromatographic and hyphenated techniques in the analysis of illegal adulteration in cosmetic products.The major content is described as follows:
1.HPLC-electrochemical detection of methotrexate and related substances in Methotrexate Injection
An HPLC-ECD method to separate and determine methotrexate and its related substances on an ODS column was established.A TSK-GEL~@ODS-100SP column (150 mm×4.6 mm,5μm)was used as analytical column with a mobile phase consisted of acetonitrile-0.2mol·L~(-1)phosphate buffer(pH6.0)(10:90).The flow rate was 1.0 mL·min~(-1),the column temperature was 35℃,and the working potential was 0.8V when using the glassy carbon disc electrode as the working electrode.The condition of electrochemical detection was studied and the difference between electrochemical detection and UV detection was compared.Methotrexate,aminopterin,and methopterin were separated.The linear range of methotrexate was 9.1-91μg·mL~(-1) (r=0.9999),the average recovery of the method was 100.4%(RSD=1.0%,n=9)。The detection limits of aminopterin,methopterin and methotrexate were 3.3,11 and 8.6ng,respectively.Compared with the UV detection method,the sensitivity of electrochemical dection is higher.It can be further employed in the detection of methotrexate in biosamples.
2.Assay and determination of related impurities in Teniposide Injection by HPLC with electrochemical detection
A sensitive analytical method for related impurities of teniposide in Teniposide Injection based on high performance liquid chromatography(HPLC)separation and electrochemical detection(ECD)has been developed.A Phenyl-Hexyl and an ODS columns were used for the separation of teniposide,lignan P,alpha-thenylidene lignan P(ATLP),picrothenylidene lignan P(PTLP)and the excipients of the Injection.The condition of electrochemical detection was investigated and the difference between EC detection and UV detection was compared.Optimal separation and detection were obtained with a mobile phase of acetonitrile-water(38:62,v/v)on an ODS column and an EC detector(carbon disc electrode as the working electrode,the working potential was set at+0.7V vs.Ag/AgCl)within 10 minutes.Good linear relationship was established between peak current and concentration of analytes in the range of 0.1-9.6μg.mL~(-1)for lignan P,0.1-9.0μg.mL~(-1)for teniposide,0.1-5.8μg.mL~(-1)for ATLP and 0.1-7.2μg.mL~(-1)for PTLP.The limit of detection(LOD,S/N=3)was 0.2ng for lignan P,1.1ng for teniposide,1.0ng for ATLP and 0.7ng for PTLP.Compared with the UV detection method,the sensitivity of EC detection is largely enhanced.It can be employed efficiently in the quality control for impurities in Teniposide Injection with good selectivity and reproducibility.
3.Determination of Entacapone and its related impurities by HPLC-ECD
An HPLC-electrochemical detection method was established for the determination of entacapone and its related impurities,z-isomer of entacapone and 3,4-dihydroxy-5-nitrobenzaldehyde.A phenyl column was used,using phosphate buffer(pH2.1) -methanol(54:46)as mobile phase.A glassy carbon disc electrode was used as the working electrode at the working potential of+0.65V.The calibration curve of entacapone,z-isomer and 3,4-dihydroxy-5-nitrobenzaldehyde were linear in the concentration range of 0.06-38.7μg.mL~(-1),0.06-42.5μg.mL~(-1)and 0.06-27.5μg.mL~(-1). The detection limit was 0.05,0.06 and.03ng,respectively.The average recovery of entacapone was 99.9%with RSD 1.5%.
4.Determination of six phenolic compounds in cosmetics by capillary electrophoresis with electrochemical detection
A method based on capillary electrophoresis with electrochemical(CE-ECD)to determine some phenolic compounds added in cosmetics was described.Effects of several factors,such as running buffer,potential applied to the working electrode, injection time and separation voltage were investigated to obtain the optimum conditions for separation and determination.With a 75 cm length of 25μm i.d. fused-silica capillary,well-defined separation of arbutin,phenol,resorcinol, hydroquinone,kojic acid and salicylic acid was achieved in the H_3BO_3-Na_2B_4O_7(40 mmol.mL~(-1))running buffer with 10 mmol.mL~(-1)SDS(pH 9.0).While operated in a wall-jet configuration,a 300μm carbon disc electrode used as the working electrode exhibits good response at+0.80V(vs.SCE)for these phenolic compounds.Good linear relationship was established between the peak current and the concentration of each analyte over 3 orders of magnitude with detection limit(S/N=3)ranging from 4.09×10~(-8)g mL~(-1)to 4.50×10~(-7)g mL~(-1).The proposed method has been successfully applied for the determination of phenolic compounds in cosmetics with satisfactory results,provides a useful monitoring method for cosmetics safety.
5.Simultaneous Determination of Antibiotics in Anti-acne Cosmetics by Rapid Liquid Chromatography with DAD
A suitable method that allows,for the first time,the simultaneous determination of nine antibiotics which may help the therapy of acne vulgaris by rapid high performance liquid chromatography with diode array detection(RPLC-DAD)in 7 minutes is presented in this work.A SB RP18(50 mm×4.6 mm;1.8μm particle size) column was used with the mobile phase consisted of a mixture of 0.1 mol.L~(-1) potassium dihydrogen phosphate(pH2.5)and acetonitrile at the gradient elution program.The correlation coefficients were all above 0.9999 in the linear range between 4-100μg mL~(-1),the average spiked recoveries(n=6)were 92.2%-103.2% with RSD ranging from 0.04%-4.5%depending on the target analytes.The method detection limits were in the range of 0.02-0.2μg mL~(-1)in anti-acne cosmetics.The analysis of real cosmetic preparations demonstrated the fitness for the whole analytical procedure.The proposed method appeared therefore as a sound alternative for official testing method,which could overcome the general problems of time consuming,lack of the specificity and precision difficulty.
6.Determination of Acrylamide residues in cosmetic products by liquid Chroma -tography with Tandem Mass Spectrometry
A liquid chromatography coupled with electrospray ionization tandem mass spectrometry(LC-MS/MS)was established to the identification and quantification of acrylamide residues in cosmetic products.The method included defatting with petroleum ether,extraction with aqueous solution of 0.1%formic acid,further liquid-liquid extraction with petroleum ether.Acrylamide was detected with an Atlantis C18 column(2.1×150 mm,3μm)using 0.1%formic acid in water/0.1% formic acid in methanol(98:2)as mobile phase A,0.1%formic acid in methanol as mobile phase B in gradient elution mode.The analytical method in the present study was well verified and good results were obtained with respect to precision, repeatability and spiked recovery.
本论文采用高效液相色谱法(HPLC)-电化学检测(ECD)联用技术对药物的杂质进行了分离测定,比较了HPLC-ECD和HPLC-UV的区别。分别采用快速液相色谱法(RRLC)-光电二极管阵列检测器(DAD)联用技术,高效毛细管电泳(HPCE)-电化学检测(ECD)联用技术,高效液相色谱法(HPLC)-串联质谱检测(MS/MS)联用技术测定了化妆品中的限用禁用药物,主要内容如下:
1.甲氨蝶呤注射液含量及有关物质的HPLC-ECD法分离测定研究
以乙腈-0.2mol·L~(-1)磷酸盐缓冲液(pH6.0)(10∶90)为流动相,采用TSK-GELODS-100SP色谱柱(150mm×4.6mm,5μm),流速为1.0mL·min~(-1),柱温35℃时,在+0.8V电位处,以玻碳圆盘电极为工作电极,银-氯化银电极为参比电极,建立了以高效液相色谱电化学检测对甲氨蝶呤及其有关物质进行分离分析的方法。甲氨蝶呤的线性范围为9.1-91μg·mL~(-1),r=0.9999;平均回收率为100.4%,RSD为1.0%(n=9)。氨蝶呤、甲蝶呤和甲氨蝶呤的最低检出量分别为3.3、11和8.6ng。电化学安培检测与紫外检测相比,具有较高的灵敏度,能有效地应用于甲氨蝶呤注射液中甲氨蝶呤含量以及极微量有关物质的检测。
2.替尼泊苷注射液含量及有关物质的HPLC-ECD法分离检测研究
建立以高效液相色谱电化学检测对替尼泊苷及其有关物质进行分离分析的方法。以PhenomenexR LUNA Phenyl-Hexyl柱,以乙腈-水(38∶62)为流动相,流速为1.0mL.min~(-1),玻碳圆盘电极为工作电极,银-氯化银电极为参比电极,在+0.7V电位处,实现了替尼泊苷及其有关物质与注射液中各种赋形剂的分离分析。木酚素、替尼泊苷、α-亚乙基木酚素P和苦亚乙基木酚素P分别在0.1-9.6μg.mL~(-1),0.1-9.0μg.mL~(-1),0.1-5.8μg.mL~(-1)和0.1-7.2μg.mL~(-1)浓度范围内呈良好线性,检测限(LOD,S/N=3)分别为0.2ng,1.1ng,1.0ng和0.7ng。电化学安培检测与紫外检测相比,具有较高的灵敏度和选择性,能有效地应用于替尼泊苷注射液中有关物质的检测和含量的测定。
3.恩他卡朋含量及有关物质的HPLC-ECD法分离测定研究
建立以高效液相色谱-电化学检测对恩他卡朋及其有关物质(z-异构体和3,4-二羟基-5-硝基苯甲醛)进行分离分析的方法。采用苯基柱,以甲醇-磷酸盐缓冲溶液(pH 2.1)(46∶54)为流动相,玻碳圆盘电极为工作电极,银-氯化银电极为参比电极,在+0.65V工作电位处,实现了恩他卡朋及其有关物质的分离分析。恩他卡朋在0.06～38.6μg mL~(-1)、z-异构体在0.06～42.5μg mL~(-1)、3,4-二羟基-5-硝基苯甲醛在0.06～27.5μg mL~(-1)浓度范围内线性关系良好,检测限分别为0.05、0.06和0.03ng。恩他卡朋的平均回收率为99.9%,RSD为1.5%。电化学安培检测具有较高的灵敏度和选择性,能有效地应用于恩他卡朋原料和片剂中有关物质的检测和片剂含量的测定。
4.胶束毛细管电泳安培检测法分离测定化妆品中限用禁用酚类化合物
采用毛细管电泳-电化学检测技术(CE-ECD)同时测定了化妆品中限用禁用酚类化合物。考察了工作电极的氧化电位、运行缓冲液的酸度和浓度、分离电压、进样时间等因素对分离和检测的影响。在优化条件下,以直径300μm的碳圆盘电极为工作电极,工作电位为+0.80 V(vs.SCE),在40 mmol L~(-1)pH9.0的H_3BO_3-Na_2B_4O_7缓冲液(含10 mmol L~(-1) SDS)中,上述组分实现了较好分离。被测物浓度与峰电流在三个数量级范围内呈良好线性关系,检测限在4.09×10~(-8)g.mL~(-1)到4.50×10~(-7)g.mL~(-1)之间,该方法简单可靠,已成功应用于不同化妆品中限用禁用酚类化合物的测定,是一种有效的化妆品安全分析检测方法。
5.RPLC-DAD法分离测定化妆品中抗生素
建立了祛痘除螨类化妆品中8种抗生素及甲硝唑同时测定的快速高效液相色谱分析方法。以Agielent1200快速高效液相色谱,采用SB和XDB-C18(1.8μm,4.6mm i.d.×50 mm)色谱柱,0.1 mol L~(-1)磷酸二氢钾溶液(用磷酸调节pH为2.5)-乙腈为流动相进行梯度洗脱,在7分钟内首次实现了8种抗生素及甲硝唑的同时分离测定,常规液相色谱分离至少需要30分钟才能完成一次分析。在4～100μg.mL~(-1)线性范围内,9种物质的线性相关系数均＞0.9999,最低检出限为0.25～1.4ng。在低中高三个添加水平,9种物质的平均回收率为92.2%～103.2%,相对标准偏差为0.04%～4.5%。并考察了不同状态的化妆品样品的提取方式。该法操作简便、准确、快速,能够检测祛痘除螨类化妆品中8种抗生素以及甲硝唑的含量。
6.HPLC-MS/MS测定化妆品中丙烯酰胺残留单体
采用高效液相色谱-串联质谱,Atlantis C18色谱柱(2.1×150 mm,3μm),以0.1%甲酸溶液-0.1%甲酸甲醇溶液(98∶2)为流动相A,0.1%甲酸甲醇溶液为流动相B进行梯度洗脱。添加同位素内标~(13)C3-丙烯酰胺于样品中,经0.1%甲酸溶液提取,石油醚液液萃取净化,串联质谱多反应监测模式,以保留时间及质谱图定性,以峰高或峰面积进行定量,测定化妆品中丙烯酰胺单体残留量。采用本方法的检出限为0.01 ng,最低检出浓度为0.01mg/Kg。方法适用于化妆品中丙烯酰胺单体残留量的测定,方法灵敏度高、重现性好。
High performance liquid chromatography(HPLC)and high performance capillary electrophoresis(HPCE)are the most important separation techniques in liquid separation method.The principle,character,and key factors of HPLC and HPCE technologies were described.The history,current situation and future trends of HPLC and HPCE were reviewed in this thesis.HPLC has greatly evolved into a major analytical technique that is routinely employed for the rapid,reliable and highly sensitive determination of numerous compounds in pharmaceutical analysis and quality control of cosmetic products.While HPCE also plays an important role in determination of impurities of pharmaceutical analysis,in chiral separation,as well as in illegal adulteration in cosmetic products,due to its speed,efficiency,reproducibility, ultra-small sample volume and ease of clearing up the contaminants.This review covers around 269 references including all types of chromatographic and hyphenated techniques reported in the latest two years.
The present research work describes the studies of high performance liquid chromatography with electrochemical detection in the quality control of impurities in pharmaceutical preparations,and different types of chromatographic and hyphenated techniques in the analysis of illegal adulteration in cosmetic products.The major content is described as follows:
1.HPLC-electrochemical detection of methotrexate and related substances in Methotrexate Injection
An HPLC-ECD method to separate and determine methotrexate and its related substances on an ODS column was established.A TSK-GEL~@ODS-100SP column (150 mm×4.6 mm,5μm)was used as analytical column with a mobile phase consisted of acetonitrile-0.2mol·L~(-1)phosphate buffer(pH6.0)(10:90).The flow rate was 1.0 mL·min~(-1),the column temperature was 35℃,and the working potential was 0.8V when using the glassy carbon disc electrode as the working electrode.The condition of electrochemical detection was studied and the difference between electrochemical detection and UV detection was compared.Methotrexate,aminopterin,and methopterin were separated.The linear range of methotrexate was 9.1-91μg·mL~(-1) (r=0.9999),the average recovery of the method was 100.4%(RSD=1.0%,n=9)。The detection limits of aminopterin,methopterin and methotrexate were 3.3,11 and 8.6ng,respectively.Compared with the UV detection method,the sensitivity of electrochemical dection is higher.It can be further employed in the detection of methotrexate in biosamples.
2.Assay and determination of related impurities in Teniposide Injection by HPLC with electrochemical detection
A sensitive analytical method for related impurities of teniposide in Teniposide Injection based on high performance liquid chromatography(HPLC)separation and electrochemical detection(ECD)has been developed.A Phenyl-Hexyl and an ODS columns were used for the separation of teniposide,lignan P,alpha-thenylidene lignan P(ATLP),picrothenylidene lignan P(PTLP)and the excipients of the Injection.The condition of electrochemical detection was investigated and the difference between EC detection and UV detection was compared.Optimal separation and detection were obtained with a mobile phase of acetonitrile-water(38:62,v/v)on an ODS column and an EC detector(carbon disc electrode as the working electrode,the working potential was set at+0.7V vs.Ag/AgCl)within 10 minutes.Good linear relationship was established between peak current and concentration of analytes in the range of 0.1-9.6μg.mL~(-1)for lignan P,0.1-9.0μg.mL~(-1)for teniposide,0.1-5.8μg.mL~(-1)for ATLP and 0.1-7.2μg.mL~(-1)for PTLP.The limit of detection(LOD,S/N=3)was 0.2ng for lignan P,1.1ng for teniposide,1.0ng for ATLP and 0.7ng for PTLP.Compared with the UV detection method,the sensitivity of EC detection is largely enhanced.It can be employed efficiently in the quality control for impurities in Teniposide Injection with good selectivity and reproducibility.
3.Determination of Entacapone and its related impurities by HPLC-ECD
An HPLC-electrochemical detection method was established for the determination of entacapone and its related impurities,z-isomer of entacapone and 3,4-dihydroxy-5-nitrobenzaldehyde.A phenyl column was used,using phosphate buffer(pH2.1) -methanol(54:46)as mobile phase.A glassy carbon disc electrode was used as the working electrode at the working potential of+0.65V.The calibration curve of entacapone,z-isomer and 3,4-dihydroxy-5-nitrobenzaldehyde were linear in the concentration range of 0.06-38.7μg.mL~(-1),0.06-42.5μg.mL~(-1)and 0.06-27.5μg.mL~(-1). The detection limit was 0.05,0.06 and.03ng,respectively.The average recovery of entacapone was 99.9%with RSD 1.5%.
4.Determination of six phenolic compounds in cosmetics by capillary electrophoresis with electrochemical detection
A method based on capillary electrophoresis with electrochemical(CE-ECD)to determine some phenolic compounds added in cosmetics was described.Effects of several factors,such as running buffer,potential applied to the working electrode, injection time and separation voltage were investigated to obtain the optimum conditions for separation and determination.With a 75 cm length of 25μm i.d. fused-silica capillary,well-defined separation of arbutin,phenol,resorcinol, hydroquinone,kojic acid and salicylic acid was achieved in the H_3BO_3-Na_2B_4O_7(40 mmol.mL~(-1))running buffer with 10 mmol.mL~(-1)SDS(pH 9.0).While operated in a wall-jet configuration,a 300μm carbon disc electrode used as the working electrode exhibits good response at+0.80V(vs.SCE)for these phenolic compounds.Good linear relationship was established between the peak current and the concentration of each analyte over 3 orders of magnitude with detection limit(S/N=3)ranging from 4.09×10~(-8)g mL~(-1)to 4.50×10~(-7)g mL~(-1).The proposed method has been successfully applied for the determination of phenolic compounds in cosmetics with satisfactory results,provides a useful monitoring method for cosmetics safety.
5.Simultaneous Determination of Antibiotics in Anti-acne Cosmetics by Rapid Liquid Chromatography with DAD
A suitable method that allows,for the first time,the simultaneous determination of nine antibiotics which may help the therapy of acne vulgaris by rapid high performance liquid chromatography with diode array detection(RPLC-DAD)in 7 minutes is presented in this work.A SB RP18(50 mm×4.6 mm;1.8μm particle size) column was used with the mobile phase consisted of a mixture of 0.1 mol.L~(-1) potassium dihydrogen phosphate(pH2.5)and acetonitrile at the gradient elution program.The correlation coefficients were all above 0.9999 in the linear range between 4-100μg mL~(-1),the average spiked recoveries(n=6)were 92.2%-103.2% with RSD ranging from 0.04%-4.5%depending on the target analytes.The method detection limits were in the range of 0.02-0.2μg mL~(-1)in anti-acne cosmetics.The analysis of real cosmetic preparations demonstrated the fitness for the whole analytical procedure.The proposed method appeared therefore as a sound alternative for official testing method,which could overcome the general problems of time consuming,lack of the specificity and precision difficulty.
6.Determination of Acrylamide residues in cosmetic products by liquid Chroma -tography with Tandem Mass Spectrometry
A liquid chromatography coupled with electrospray ionization tandem mass spectrometry(LC-MS/MS)was established to the identification and quantification of acrylamide residues in cosmetic products.The method included defatting with petroleum ether,extraction with aqueous solution of 0.1%formic acid,further liquid-liquid extraction with petroleum ether.Acrylamide was detected with an Atlantis C18 column(2.1×150 mm,3μm)using 0.1%formic acid in water/0.1% formic acid in methanol(98:2)as mobile phase A,0.1%formic acid in methanol as mobile phase B in gradient elution mode.The analytical method in the present study was well verified and good results were obtained with respect to precision, repeatability and spiked recovery.
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