Determination of Benzo[a]pyrene in Edible Oil by High Performance Liquid Chromatography-Fluorescence Detector (HPLC-FL)

In this study, an optimized high performance liquid chromatography-fluorescence detector (HPLC-FL) method for the determination of benzo[a]pyrene in edible oil was established. HPLC was performed with Thermo Fisher Scientific C18 column (250 mm×4.6 mm, 5 μm) as the chromatographic column and acetonitrile and water as the mobile phase, and the excitation wavelength and emission wavelength of fluorescence detector were 286 and 430 nm, respectively. The response was high, and the linear range was 0.5-10.0 ng/ml. The lowest limit of detection was 0.11 ng/ml, and the average recovery was 92.5%. This method is suitable for quantitative analysis of benzo[a]pyrene content in edible oil.

Research Advances in Detection Techniques of High Performance Liquid Chromatography-Refractive Index Detector

As a highly sensitive and stable detector,refractive index detector is usually used for quantitative detection of substances such as polymer,sugar and organic acid. The research reviewed the application of HPLC-RID in the fields of quantitative determination of medicine and food,in order to lay a foundation for wider use of RID.

Determination of AF and AFG in Red Ginseng by High Performance Liquid Chromatography with Evaporative Light Scattering Detector (HPLC-ELSD)

[Objective]The paper was to establish a method for determining AF and AFG in red ginseng.[Method]A new simple,rapid and sensitive method for simultaneous determination of two amadori compounds,arginyl-fructose(AF)and arginyl-fructosyl-glucose(AFG),in extracts of three kinds of ginseng preparations was developed and validated using high performance liquid chromatography with evaporative light scattering detector(HPLC-ELSD).Two target analytes were efficiently separated by Prevail CTM18 column(4.6 mm×250 mm,5μm)at the flow rate of 0.8 mL/min within 15 min of single chromatographic run.[Result]Under optimized conditions,the detection limits were 0.015 and 0.02 mg/mL for AF and AFG,respectively.Calibration curves of peak area for two analytes were linear over three orders of magnitude with the correlation coefficients greater than 0.999.The average recoveries,precision,reproducibility and stability for two analytes(AF and AFG)were 99.5% and 100.9%,0.43% and 0.47%,0.46% and 0.43%,0.41% and 0.49%,respectively.[Conclusion]This method was successfully applied for quantifying AF and AFG in red ginseng and the method was efficient,sensitive and accurate.

Quantification of six bioactive compounds in Zhenqi Fuzheng preparation by high-performance liquid chromatography coupled with diode array detector and evaporative light scattering detector

A simple and accurate high-performance liquid chromatography（HPLC）coupled with diode array detector（DAD）and evaporative light scattering detector（ELSD）was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation（ZFP）.The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis（HCA）,which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Fast determination of tobramycin by reversed-phase ion-pair high performance liquid chromatography with a refractive index detector

A simple and direct method without a deriva- tion step for routine analysis of tobramycin has been developed. This method used reversed-phase ion-pair high performance liquid chromatography （HPLC） with a refractive index （RI） detector and a C18 column which is stable at pH above 1.00. The presence of 4.50mg.mL ] trifluoroacetic acid （TFA） in the mobile phase improved the protonation of tobramycin and the formation of ionpairs, and thus reduced its hydrophility. This unique separation-detection combination showed good linearity with correlation coefficients 0.9996 in the concentration range of 0.25-2.50mg.mL 1. The quantitation limit and detection limit were determined to be 0.23 mg.mL ~ and 0.071 mg. mL ~, respectively. Tobramycin was recovered in 98.00%, 98.84% and 99.64% for tobramycin solutions at concentrations of 2.25 mg.mL^-1, 1.50mg.mL 1 and 0.75 mg.mL ＇, respectively. The relative standard deviations for six spiked samples ranged from 0.20% to 2.40%, indicating a good reproducibility of this method.

PEAK IDENTIFICATION FROM INTERACTION INDEX IN REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

The interaction index c which was derived from the fundamental retention equationlogk’ =a + cC_B in reversed--phase high--performancc liquid chromatography (RP-HPLC) quan-titatively describes the difference in the interaction between solute--strong. solvent andsolute--weak solvent; it has shown to be a constant for a specific solute even when columnsystems with different C18 packings are used. The theoretical basis for peak identificationby using interaction index has been proposed, which was based on the a,c values on stan-dard C18 column by utilizing linear a-a plots on column pairs and the linear relationship be-tween parameters a and c for the structural related compounds. Through the establishmentof parameters a,c data based on the standard C18 column for a certain type of compounds,the retention of thesc compounds on various C18 columns can be predicted. Typical exam-ples have been given to verify the correctness of this method.

丹磺酰肼衍生-高效液相色谱-荧光检测器法测定水中痕量3-羟基丙醛

提出了丹磺酰肼衍生-高效液相色谱-荧光检测器法测定水中痕量3-羟基丙醛(3-HPA)的方法。将1.0 g水样、2.0 mL含200 mg·L^(-1)丹磺酰肼的乙腈溶液、1.0 mL 3.0%(体积分数)乙酸溶液混合,再用乙腈定容至50 mL,于40℃衍生反应30 min。以Agilent Eclipse Plus C_(18)色谱柱为固定相,以不同体积比的乙腈-0.10%(质量分数)七氟丁酸溶液的混合溶液为流动相进行梯度洗脱,采用荧光检测器测定。结果表明:衍生试剂丹磺酰肼与3-HPA衍生物在20 min内可实现基线分离;3-HPA的质量浓度在7.6~380.0μg·L^(-1)内与衍生物的峰面积呈线性关系,检出限(3S/N)为1.1μg·L^(-1);方法用于实际水样分析,测定值的相对标准偏差(n=6)为0.71%,3-HPA的加标回收率为98.0%~102%。

高效液相色谱-蒸发光散射法测定油莎豆块茎发育过程中的水溶性糖

基于高效液相色谱-蒸发光散射法(HPLC-ELSD)建立油莎豆块茎发育过程中水溶性糖(果糖、葡萄糖、蔗糖)含量的检测方法。以乙腈∶水(体积比75∶25)为流动相,流速为1.0 mL/min,并将蒸发光散射检测器条件优化为氮气压力206.8 kPa、漂移管温度65℃。结果表明:3种水溶性糖在0.10~20.0 mg/mL质量浓度范围内线性关系良好(r>0.999),检出限为0.04~0.10 g/100 g,回收率为96.0%~100.2%,相对标准偏差(RSD)为1.00%~7.32%(n=6)。在油莎豆块茎发育过程中,其果糖含量为0.376~0.692 g/100 g,葡萄糖含量为0.366~0.787 g/100 g,两者呈显著正相关(P<0.01);蔗糖含量为5.49~24.2 g/100 g,呈先降低后总体升高的趋势。所建方法灵敏度高、重现性好,适用于油莎豆块茎发育过程中水溶性糖的测定。

HPLC-ELSD同时测定腺梗豨莶草中6个二萜成分的含量

为了研究腺梗豨莶草中6个二萜成分(对映海松烯型二萜:奇任醇、Hythiemoside B和豨莶精醇;对映贝壳杉烷型二萜:对映-17,18-二羟基-贝壳杉烷-19-羧酸、对映-16β,17-二羟基-贝壳杉烷-19-羧酸和对映-16α氢-贝壳杉烷-17,19-二羧酸)的含量,本试验建立了高效液相色谱法-蒸发光散射检测器(HPLC-ELSD)同时测定二萜类化合物含量。样品粉碎过筛加甲醇和乙酸乙酯回流提取,蒸发减压除去溶剂,甲醇溶解,0.45μm滤膜滤过,取续滤液进行测定。色谱条件:色谱柱为Waters Symmetry Shield^(TM)RP18柱(250 mm×4.6 mm,5μm),流动相为0.3%甲酸水溶液-乙腈(v/v),梯度洗脱,流速为1.0 mL/min。蒸发光散射检测器漂移管温度为103℃,雾化气流速为3.0 L/min。应用该方法测定腺梗豨莶草样品不同部位中6个二萜类化合物的含量,同时比较叶、枝、茎中对映海松烯型二萜、对映贝壳杉型二萜和总二萜含量的差异。结果显示,6个二萜成分在其线性范围内线性关系良好(r≥0.9992);日内和日间精密度相对标准偏差(RSD)均小于3.5%;回收率介于96.5%~101.5%,RSD均小于2.3%;腺梗豨莶草不同部位(叶、枝、茎)的二萜类化合物含量差异较大。结果表明,本试验所建立的HPLC-ELSD方法简便、准确、重复性好,为腺梗豨莶草药材全面的质量评价和临床应用中最佳药用部位的选择提供了参考。

芪归养血颗粒质控标准研究

目的研究建立芪归养血颗粒质控的标准。方法采用薄层色谱法(TLC)对黄芪、淫羊藿、菟丝子进行定性鉴别,通过超高效液相色谱-蒸发光散射检测器(UPLC-ELSD)测定制剂中黄芪甲苷含量:色谱柱:Acclaim RSLC 120 C_(18)(100 mm×2.1 mm,2.2µm),ELSD,流动相为乙腈-水(32∶68),流速为0.4 ml/min;柱温30℃;进样体积为1μl。结果TLC能特异性鉴别黄芪、淫羊藿、菟丝子。黄芪甲苷的注射量与峰面积具有良好的线性关系,线性范围为0.0826~0.8260µg,r=0.9996;平均回收率为99.19%,相对标准偏差(RSD)为0.74%。结论采用TLC法和UPLC-ELSD检测对芪归养血颗粒分别进行了定性鉴别和定量分析,所建立的方法专属性强,结果准确,可作为芪归养血颗粒的质控方法进行应用。

市售豉香型白酒品质及风味感官分析

目的探究市售豉香型白酒品质特性,通过理化指标与感官风味评价的总体分析,构建其之间的相互关系。方法以12种市售豉香型白酒为研究对象,采用高效液相色谱法(high performance liquid chromatography,HPLC)结合风味感官评价方法,通过有机酸含量、酒精度、总酸、总酯、酸酯总量、pH及风味感官特性等指标综合分析市售豉香型白酒的品质。结果12种市售豉香型白酒中均检测出乳酸和乙酸2种有机酸,并未检测出草酸和柠檬酸;相比于浓香、清香等白酒,豉香型白酒中苹果酸和酒石酸含量较低;12种市售豉香型白酒属于平均为29.8%vol的低度酒,总酸和总酯含量平均分别为0.54 g/L和0.53 g/L;92%的样品酸酯总量不小于8mmol/L,符合一级豉香型白酒的要求;样品的pH都在3.06~4.53内;市售豉香型白酒在水果香、蜜香、米香等风味强度上无显著性差异,而在醇香和脂肪味上具有显著差异。结论豉香型白酒相关理化指标所代表的风味物质对其风味感官具有一定贡献,研究结果为豉香型白酒理化指标与风味感官存在相互关系,理化指标测定结果对豉香型白酒风味感官评价具有导向作用。

Phytochemical Analysis of Ziziphus jujuba Leaves in Six Cultivars at the Whole Life Stage by High Performance Liquid Chromatography

Leaves of Ziziphus jujuba Mill.（Z. jujuba） were used as a folk medicine to treat children suffering from typhoid fever, furuncle and ecthyma in China. The present study was to establish a high performance liquid chroma- tography-diode array detector（HPLC-DAD） method for quality control of Z. jujuba leaves. The validated method was applied to the simultaneous characterization and quantification of sixteen main constituents in thirty samples, which comprised the whole life stage of Z. ]ujuba leaves from six cultivars in Yulin, Shannxi Province, China. HPLC fin- gerprint file of Z. jujuba leaves was set up and the results indicate that the major constituents of Z. jujuba leaves are （-）-catechin（1）, quercetin-3-O-robinobioside（3）, rutin（4） and quercetin-3-O-a-L-arabinosyl-（1---~2）-a-L-rhamnoside（6） with the total contents of the nine flavonoids in respective batch ranging from 18.52 mg/g to 39.51 mg/g. In addition, the contents of compounds depend on both the season and the cultivar. During ripening of Z. jujuba leaves, flavo- noids of all the cultivars present a similar trend： initially decrease from 25th, Jun. to 5th, Aug., 2015, and then in- crease for a period of time, finally slightly fluctuate on 5th Oct. The highest contents of total flavonoids were regis- tered on 25th Jun., 2015. Different cultivars have 50%--90% variation on the content for the three classes of com- pounds. Taken together, the results demonstrate that Z. jujuba leaves have great potential to be a new health-promoting resource based on flavonoids and the best sampling time of them is in June.

高效液相色谱-示差折光检测器法测定复配消毒液中3种双链季铵盐的含量

提出了高效液相色谱-示差折光检测器法测定复配消毒液中二癸基二甲基氯化铵、辛基癸基二甲基氯化铵、二辛基二甲基氯化铵等3种双链季铵盐含量的方法。复配消毒液样品经流动相稀释,以Welchrom C_(18)色谱柱为固定相,以体积比70:30的乙腈-0.01 mol·L^(-1)丁烷磺酸钠溶液的混合溶液(pH 3.5)为流动相进行洗脱,采用示差折光检测器测定3种双链季铵盐的含量。结果表明:3种双链季铵盐的质量浓度在50~1000 mg·L^(-1)内与对应的峰面积呈线性关系,检出限(3S/N)为3.3~5.1 mg·L^(-1);对实际样品进行3个浓度水平的加标回收试验,回收率为91.0%~105%,测定值的相对标准偏差(n=6)为1.3%~2.2%。采用独立样本t检验法比对本方法与高效液相色谱-蒸发光散射检测器法是否具有显著性差异,结果显示两组检测结果t值为1.85,P值为0.089,表明无显著性差异。

高效液相色谱法检测葡萄糖酸-δ-内酯

葡萄糖酸-δ-内酯常通过葡萄糖酸水溶液蒸发浓缩制备,所得产品常含有少量的葡萄糖酸和葡萄糖酸-γ-内酯。为了检测产品中葡萄糖酸-δ-内酯的质量分数,文中建立了可快速分离分析葡萄糖酸-δ-内酯、葡萄糖酸和葡萄糖酸-γ-内酯的高效液相色谱法,并优化了葡萄糖酸-δ-内酯的分析条件。以SPD-M20A二极管阵列检测器,采用Titank Hilic色谱柱分离葡萄糖酸-δ-内酯、葡萄糖酸和葡萄糖酸-γ-内酯,以0.01 mol/L乙腈-磷酸水溶液(体积比90∶10)为流动相进行等度洗脱,柱温为30℃,流速为1.0 mL/min,检测波长为205 nm,进样量为20μL。结果表明,葡萄糖酸-δ-内酯在7 min内可实现较好的分离;在0.5—2.5 mg/mL内具有良好的线性关系(r>0.999),检出限为0.002 mg/mL,定量限为0.5 mg/mL,3个水平的平均加标回收率为98.8%—99.9%,相对标准偏差0.1%—0.4%(n=5)。此方法分析时间短、灵敏度高、分离效果好,可用于葡萄糖酸-δ-内酯、葡萄糖酸和葡萄糖酸-γ-内酯的同时检测。

三黄洗剂指纹图谱建立及原药材相关性研究

目的建立三黄洗剂高效液相色谱法(HPLC)指纹图谱,并分析与原药材的相关性,为该制剂的质量控制提供依据。方法采用ZORBAX Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5μm),以甲醇-0.1%磷酸溶液为流动相(梯度洗脱),流速为1.0 ml/min,体积流量为1.0 ml/min,柱温为30℃,检测波长265 nm,进样量为10μl。结果建立起三黄洗剂的指纹图谱,共标定27个共有峰,指认出6个成分分别为黄芩苷、汉黄芩苷、黄芩素、小檗碱、芦荟大黄素和大黄酸,10批制剂的相似度均>0.95,与原药材的相关性良好。结论该方法简便、准确、重复性好,可以作为三黄洗剂质量控制的检测手段。

基于HPLC⁃DAD指纹图谱结合化学模式识别的红糖产地溯源方法

红糖作为一种功能食品,近年来需求量不断增长,然而市场上红糖的品质却良莠不齐,且缺乏科学的鉴别和评估方法来确保红糖的品质。该研究以我国5个红糖主产地(广西、广东、云南、贵州和香港地区)的55批红糖作为研究对象,通过构建高效液相色谱⁃二极管阵列检测器(high⁃performance liquid chromatography⁃diode array detector,HPLC⁃DAD)指纹图谱并结合共有模式法、相似度评价法和化学计量学法,建立一种能够准确追溯红糖产地的方法。研究发现,不同产地的红糖既有特有的HPLC⁃DAD指纹图谱也有共有的HPLC⁃DAD指纹图谱,特有的HPLC⁃DAD指纹图谱显示产地特有峰的保留时间具有明显的地域特性,与产地区分密切相关。产地共有指纹图谱共标定出9个共有峰,指纹图谱的相似度为0.473~0.915,表明虽然存在共有峰但共有峰峰面积存在明显差异(例如广东、广西、云南、贵州和香港地区的S2号共有峰的平均相对峰面积分别为47.592%、26.873%、814.853%、8.683%和3.111%)。共有峰峰面积的聚类热图显示不同地区的样品存在明显差异。此外,共有峰峰面积的主成分分析发现前3个主成分对产地区分的贡献率分别为59.9%、17.1%和11.6%,累计贡献率为88.6%,可以作为鉴别红糖产地的特征性指标。

荧光检测器高效液相色谱法测定樟脑丸中萘

建立高效液相色谱法测定樟脑丸中萘的含量。样品以甲醇为溶剂超声溶解,经0.22μm滤膜过滤后,以荧光检测器高效液相色谱法测定。荧光检测器激发、发射波长分别为290、330 nm,以甲醇为流动相,流量为1.0 mL/min,以Platisil ODS柱(250 mm×4.6 mm,5μm)色谱柱作为分析柱,柱温为25℃,进样体积为10μL,利用色谱峰面积外标法进行定量。该方法线性相关系数为0.9999,检出限为1.0×10^(-8)g/mL,样品加标回收率为98.4%~102.8%(n=6),测定结果的相对标准偏差为1.81%(n=6)。该方法可用于樟脑丸中萘的检测。

高效液相色谱-荧光检测器法同时测定乳及乳制品中维生素A和维生素E

目的建立高效液相色谱-荧光检测器法同时测定乳及乳制品中维生素A和维生素E的分析方法。方法样品经水解皂化、液液萃取、蒸发浓缩后,选用PFP色谱柱分离,以甲醇-水(9:1,V/V)等度洗脱,高效液相色谱-荧光检测器检测,外标法定量,计算维生素A、维生素E的含量。结果维生素A和4种维生素E异构体在0.10~5.00μg/ml范围内呈现良好线性,方法检出限为0.0100μg/g、方法定量限为0.0400μg/g,维生素A的平均回收率在85.0%~97.0%之间,RSD≤6.41%(n=6);4种维生素E异构体的平均回收率在84.0%~101%之间,RSD≤7.04%(n=6)。结论本方法简便快速,灵敏准确,可满足乳及乳制品中维生素A和维生素E的同时测定。

基于多指标成分定量、化学计量学、EW-TOPSIS分析的白术厚朴合剂质量评价研究

目的以10种成分为指标,建立白术厚朴合剂HPLC定量分析方法,联合化学计量学及熵权优劣解距离法(EW-TOPSIS)对其质量进行综合评价。方法以75%甲醇提取液为样品,采用Agilent TC-C18色谱柱,乙腈-0.1%磷酸为流动相梯度洗脱,HPLC法同时测定12批白术厚朴合剂中10个化合物(白术内酯Ⅲ、白术内酯Ⅰ、肉桂醇、肉桂酸、桂皮醛、和厚朴酚、厚朴酚、6-姜辣素、8-姜酚和10-姜酚)含量,运用化学计量学和EW-TOPSIS法对含量数据进行分析。结果经方法学验证,各指标均符合《中华人民共和国药典》要求:10种成分在各自质量浓度范围内线性关系良好,相关系数>0.999;精密度、稳定性、重复性、加样回收率RSD均<2.0%。化学计量学分析显示,前2个主成分代表10个成分92.24%的信息,12批样品聚为3类,桂皮醛、厚朴酚、和厚朴酚、白术内酯Ⅲ、6-姜辣素和白术内酯Ⅰ对白术厚朴合剂的质量影响较大。EW-TOPSIS法结果显示,12批白术厚朴合剂的最优解欧氏贴近度均值为0.2385~0.6662,各批次间质量差异较大。结论建立的多指标成分定量方法操作便捷、结果准确,可用于白术厚朴合剂的质量控制,化学计量学及EW-TOPSIS分析可实现对白术厚朴合剂质量的综合评价,为白术厚朴合剂质量标准提升提供参考。