测定食物中5种类黄酮物质的高效液相色谱法的改进

目的优化、完善分析植物性食物中5种类黄酮成分的方法。方法采用Nova-Pak C18(3.9 mm×150mm,4μm)色谱柱;流动相为0.1%三氟乙酸(A)-5%乙腈甲醇溶液(B),梯度洗脱(0～5min,90%A→60%A;5～18 min,60%A→40%A;18～21 min,40%A→10%A;21～25 min,10%A→10%A;25～30 min,10%A→90%A),流速1 ml/min;检测波长365 nm,柱温35℃。结果杨梅黄酮、槲皮素、玉米黄酮、坎二菲醇、芹菜配基5种类黄酮在0.156～20μg/ml浓度范围内与色谱峰高呈良好的线性关系,相关系数(r)为0.9995～0.9999;检测限为0.0195μg/ml(S/N≥3);低、中、高浓度的平均加标回收率(n=3)为88.5%～105.6%,相对标准偏差(RSD)均小于6.32%。结论该方法灵敏、准确,具有良好的重复性和稳定性,可用于植物性食物中5种类黄酮含量的检测。     

树莓与蓝莓中多酚类成分的HPLC测定

利用HPLC法对树莓与蓝莓中的生理活性成分进行含量测定。HPLC法采用Kromasil C18色谱柱(150 mm,4.6 mm);流动相采用色谱纯乙腈(A)-含0.1%磷酸水溶液(B)梯度洗脱,0~16 min,5%A;19 min,10%A;23 min,13%A;40 min,14%A;43 min,17%A;48 min,22%A;51 min,24%A;52 min,24%A;58 min,26%A;60 min,30%A;65 min,60%A;68 min,70%A;72 min~78 min,100%A;流速为1 mL/min;柱温30℃;检测波长为280 nm;每次进样10μL,测定树莓与蓝莓95%乙醇提取物中没食子酸、原儿茶酸、儿茶素、表儿茶素、原花青素B2、矢车菊素、鞣花酸、树莓酮、白藜芦醇、水杨酸的含量;这十种多酚类成分在三种浆果中含量高低不等,其中鞣花酸含量在黑树莓中最高(0.97μg/g),红树莓次之(0.51μg/g),蓝莓最低(0.36μg/g)。蓝莓与树莓中富含多种多酚类物质,是其具有多种生物活性的物质基础。     

超临界流体色谱—串联质谱分离和测定烟草中磷胺和速灭磷顺反异构体

  目的  快速分离和测定烟草中磷胺和速灭磷顺反异构体。  方法  通过超临界流体色谱-串联质谱(SFC-MS/MS)建立了磷胺和速灭磷顺反异构体的分析方法。  结果  结果表明:①磷胺和速灭磷顺反异构体均达到基线分离,分离时间为5 min。②在高、中、低三个不同添加浓度水平下,所有异构体的回收率在84.7%~96.7%之间,日间RSD < 5.0%,检出限(LOD)为0.6 μg/kg~1.7 μg/kg定量限(LOQ)为2.0 μg/kg~5.5 μg/kg。③采用该方法对20个烟叶实际样品进行检测,所有样品均未检出磷胺和速灭磷的异构体。  结论  该方法快速,环保,稳定可靠,可用于分离和测定烟草中磷胺和速灭磷顺反异构体的含量。      

高效液相色谱同时检测不同类型烟叶中的多种生物碱含量

为快捷准确地测定烟叶中生物碱的含量,本研究通过优化提取条件,建立了测定烟草中烟碱、新烟草碱、假木贼碱和降烟碱的高效液相色谱检测方法。优化后的检测步骤为：干烟叶和5%甲醇的料液比为100 mg/20 mL,在25℃下振荡提取30 min,色谱条件：C18柱（250 mm×4.6 mm,5 μm）,流动相A（90%水+10%甲醇,pH 6.8,0.15%醋酸钠）,流动相B（90%甲醇+10%水,pH 3.8）。烟碱在0.1~50 μg/mL、其余3种在0.005~2μg/mL的范围内,4种生物碱均具有较好的线性关系（R²>0.99）;烟碱检出限为0.000 65 μg/mL,平均回收率100.79%,RSD 3.11%;新烟草碱检出限0.088 4 μg/mL,平均回收率98.53%,RSD 3.75%;假木贼碱检出限0.034 8μg/mL,平均回收率98.70%,RSD 3.63%;降烟碱检出限0.150 9 μg/mL,平均回收率101.33%,RSD 4.61%。对4种不同类型烟叶共11个不同样品进行检测验证,检测结果可靠。该方法前处理简单快捷,灵敏度高,重复性较好,可以满足烟草样品中4种生物碱的检测要求。     

食品中虾青素的C18-HPLC-PDA分离与鉴定

在装备了二极管阵列检测器(PDA)的高压液相色谱(HPLC)上,应用C18柱,冰凉花和虾中的主要酮基类胡萝卜素-虾青素可以被分离。分离条件为:色谱柱为DiamonsilTM(5μm,4.6mm×25cm,DikmaTechnology);流动相A为乙腈-水(90∶10,V/V),用磷酸调pH3.0;流动相B为乙酸乙酯;线性梯度洗脱:B在15min内由0%增加至70%;15～25min内B维持在70%;流速=1.0mL/min;检测波长=480nm;PDA波长范围=280～580nm;进样量=20mL。根据其与参比样品的色谱行为和光谱特征比较,样品中的虾青素可被鉴定。它在C18-HPLC-PDA上的定量亦成为可能。     

高效液相色谱法测定三七类保健食品中6种皂苷的含量

目的建立高效液相色谱法(high performance liquid chromatography,HPLC)测定三七类保健食品中三七皂苷R_1、人参皂苷Rg_1、人参皂苷Rg_2、人参皂苷Rb_1、人参皂苷Rd和人参皂苷Re含量的分析方法。方法用Kromasil 100-5-C_(18)(4.6 mm×250 mm,5μm)色谱柱,以乙腈(A)-水(B)为流动相,梯度洗脱(0~25 min,17%A→19%A;25~45 min,19%A→29%A;45~55 min,29%A;55~95 min,29%A→40%A;95~97 min,40%A;97~100 min,40%A→17%A),流速1.0 mL/min,检测波长203 nm,柱温40℃。结果三七皂苷R_1、人参皂苷Rg_1、Re、Rg_2、Rb_1、Rd分别在4.060~203.0,4.818~240.9,4.154~207.7,4.282~214.1,3.894~194.7,4.124~206.2μg/mL浓度范围内线性关系良好,相关系数都在0.9985以上。该方法 6种皂苷平均回收率均高于93.4%,相对标准偏差(relative standard deviation,RSD)小于2.5%。结论本法准确可靠、灵敏度高、重现性好,可作为含三七类保健食品的质量控制方法。     

RP-HPLC法测定烟草中的质体色素

为了快速测定烟草及其制品中的质体色素,考察了萃取溶剂、萃取方式、萃取时间、流动相、柱温和柱流速对测定结果的影响,建立了同时测定烟草样品中6种质体色素的方法,并采用该法测定了10种烟草样品中的质体色素.结果表明:①烟草样品中的色素用90%丙酮溶液超声提取20 min效果较好;②用Waters Nova-Pak-C18作色谱柱,异丙醇和乙腈-水(体积比80:20)为流动相,梯度洗脱,流速0.5mL/min,6种色素均能完全分离;③该法的回收率为96.7%-110.7%,RSD均小于5%;④2种新鲜的冻干烟叶样品中均检测出了新黄质、紫黄质、叶黄素、叶绿素b、叶绿素a、B-胡萝卜素等色素,且其含量较高,而原烟及卷烟烟丝中仅检测出了叶黄素和β-胡萝卜素,且其含量也较新鲜烟叶低得多.该方法适合批量烟草样品中这6种质体色素的快速分析.     

高效液相色谱法同时测定心动颗粒中阿魏酸和欧前胡素的含量

目的建立高效液相色谱法(HPLC)同时测定心动颗粒中阿魏酸和欧前胡素含量。方法色谱柱为Welchrom@C18反相色谱柱(4.6 mm×250 mm,5μm);流动相为乙腈(A)-水(10 mmol/L磷酸)(B),梯度洗脱:0~3.5 min,15%A;3.5~12 min,15%A→25%A;12~21 min,25%A);流速1.0 m L/min;检测波长310 nm;柱温35℃。结果阿魏酸和欧前胡素的质量浓度分别在5.19~51.90μg/m L和5.20~52.05μg/m L浓度范围内呈良好的线性关系(r20.9995),平均回收率分别为98.34%和98.06%,RSD分别为1.02%和0.96%(n=6)。结论本测定方法简便、准确,稳定性、重复性好,适用于心动颗粒中阿魏酸和欧前胡素的含量测定。     

HPLC同时测定木香顺气丸中5种成分的含量

目的建立同时测定木香顺气丸中5种活性成分含量的方法。方法采用高效液相色谱法(HPLC),色谱柱为Kromasil C18柱(250 mm×4.6 mm,5μm),以乙腈为流动相A,0.2%磷酸溶液为流动相B,梯度洗脱:0~5 min,10%A;5~20 min,10%→20%A;20~40 min,20%→60%A;40~60 min,60%A;分段设定检测波长:0~20.0 min为210 nm(木香烃内酯、去氢木香内酯);20.0~30.0 min为292 nm(和厚朴酚);30.0~45.0 min为211 nm(厚朴酚);45.0~60.0 min为250 nm(甘草酸),流速1.0 m L/min,柱温30℃,进样量20μL。结果香烃内酯、去氢木香内酯、和厚朴酚、厚朴酚和甘草酸分离度良好,进样量分别在0.025~1.008,0.031~1.232,0.022~0.864,0.058~2.304,0.045~1.800μg范围内线性关系良好,r均大于0.9990,平均加样回收率为99.2%~99.7%(RSD=0.74%~1.05%)。5批样品中香烃内酯、去氢木香内酯、和厚朴酚、厚朴酚和甘草酸含量分别在6.17~6.51,2.74~2.89,13.2~13.66,2.54~2.87,4.40~4.65 mg/g。结论本方法专属性好、灵敏度高、准确度高,为木香顺气丸的质量控制提供依据。     

HPLC同步测定秃疮花中紫堇丁碱同分异构体的含量

研究HPLC同步测定秃疮花中紫堇丁碱同分异构体含量的方法。采用工业乙醇超声辅助法提取秃疮花全草干粉制得乙醇提取物,用等量石油醚和氯仿溶剂依次萃取,氯仿相采用柱层析法同步分离得到紫堇丁碱同分异构体,采用HPLC测定其含量。色谱条件:采用HPLC(E2695 Waters,1525-2707 TCM-2489)、柱型C₁₈(250 mm×4.6mm,5μm),甲醇(A)-水(B)为流动相,梯度洗脱(0~5 min,10%A;5~35 min,10%A→100%A;45~48 min,100%A→10%A,48~50 min,10%A),上样量1 mL,进样体积10μL,每个样品运行检测时间50 min;柱温30℃,检测波长260nm。结果表明,秃疮花中紫堇丁碱含量为0.79%,异紫堇丁碱含量为1.33%。紫堇丁碱和异紫堇丁碱是秃疮花酸化氯仿相的主要活性成分,其同步富集工艺及HPLC同时检测方法未见报道,试验为定量测定药食同源植物中单体化合物的含量奠定试验基础和理论依据。     

烟叶及提取物中茄尼醇的高效液相色谱标准化测定方法研究

为了更好地理解和应用笔者制定的国家标准《烟叶和烟叶提取物中茄尼醇的测定－高效液相色谱法》,笔者详细说明了该方法的技术要点,筛选出最佳的提取条件和皂化条件。高效液相色谱检测条件为;采用Waters Atlantis T3- C₁₈柱(4.6×150 mm,3 μm),柱温35 ℃;以异丙醇-甲醇(体积比为30:70)为流动相,流速为1.0 mL/min;采用二极管阵列检测器,检测波长213 nm。实验结果表明,该方法对茄尼醇的检出限为1 mg/kg,在0.1～5.0 g/100g线性范围内,茄尼醇含量与峰面积的相关系数达0.9999以上;方法加标回收率达到95%以上;相对标准偏差小于5%(n=6)。该方法快速、简便、具有良好的精密度和准确性。     

A simple,sensitive HPLC-DAD method for simultaneous determination of carotenoids,chlorophylls and α-tocopherol in leafy vegetables

Leafy vegetables are the important components of our diet and are the source of several beneficial phytochemicals. A sensitive, simple analytical method is therefore required to precisely measure the phytochemical composition. A validated reversed phase high performance liquid chromatography with diode array detection (HPLC-DAD) method was developed to determine simultaneously carotenoids, chlorophylls and α-tocopherol composition of six leafy vegetables (B. compestris, B. rapa, B. juncea, M. neglecta, and two spinach varieties). Carotenoids were extracted and efficiently separated using a tertiary mobile gradient system of methanol–water, water and MTBE in 40 min on a reversed phase C18 column. The method was simple, precise, accurate and highly reproducible. Twelve carotenoids namely lutein and its three isomers, β-carotene-5,6-epoxide, neoxanthin, violaxanthin, two cis-isomers of zeaxanthin, 8-apo-carotenal, all-trans-β-carotene and its 13-cis-isomer; one fatty acid ester (β-cryptoxanthin ester); α-tocopherol and chlorophyll a & b were quantified in vegetable leaves. α-Tocopherol, neoxanthin, violaxanthin, lutein, 8-apo-carotenal, chlorophyll a and all-trans-β-carotene were present in higher amounts. Significant variations in the major compounds were observed in the selected vegetables. It was concluded that the developed method was highly sensitive, accurate and can be used to analyze carotenoids, chlorophylls and α-tocopherol simultaneously in leafy vegetables as well as in other plant leaves.     

特殊烟叶烘烤过程中生理生化变化及烤后质量特点

以正常烟叶为对照,对黑暴烟、黑胫病烟叶、叶斑病烟叶、病毒病烟叶等特殊烟叶烘烤过程中的生理生化指标及烤后烟叶质量进行了分析研究。结果表明,特殊烟叶烘烤前中期水分含量偏高或偏低;黑暴烟烘烤过程中叶绿素和类胡萝卜素含量都明显高于正常烟叶和其他特殊烟叶,类胡萝卜素/叶绿素（类叶比）变化较小,黑胫病、病毒病、叶斑病烟叶烘烤过程中类叶比的高峰值出现时间早于正常烟叶;烘烤前期特殊烟叶多酚氧化酶活性比正常烟叶高;特殊烟叶烤后碳水化合物含量低,含氮化合物含量高,评吸时香吃味变差、杂气和刺激性加重。     

诱导子对烟草植株叶片酶活性及香气物质积累的效应

采用盆栽试验,通过叶面喷施诱导子,考察了诱导子对烟草次生代谢关键酶活性及香气物质积累的影响.结果表明,经诱导子处理过的烟草叶片苯丙氨酸解氨酶、多酚氧化酶和过氧化物酶活性均高于对照,且腺毛分泌物、石油醚提取物、类胡萝卜素、多酚类物质含量也高于对照.综合考虑,100 mg/L浓度的茉莉酸甲酯诱导子和200 mg/L浓度的壳聚糖诱导子处理烟叶较有利于烟草香气成分的生成.     

不同品种烤烟多酚含量和组成的差异分析

采用反相高效液相色谱法对2005和2006年河南中部烟区种植的4个烤烟品种初烤烟叶中的6种多酚成分进行了测定。结果表明:①不同烤烟品种的多酚类物质含量存在不同程度的差异,且两年的情况基本相同,中烟100的多酚类物质含量较高,云烟85和云烟87的多酚类物质含量相对较低;②绿原酸及其同分异构体和芸香苷占烤烟多酚类物质的90%以上,是主要的多酚类物质,山奈酚糖苷和莨菪亭含量极低,中烟100的总绿原酸、绿原酸占多酚总量的比例高于NC89和云烟系列,云烟85和云烟87的芸香苷占多酚总量的比例明显高于其它品种;③不同部位多酚含量的差异与烤烟品种也有一定关系。     

烤后烟叶类胡萝卜素的稳定性及其影响因素

为了探明烤后烟叶褪色的成因,进行了光照、温度、相对湿度、酸、碱、金属离子和还原剂等影响烤后烟叶中类胡萝卜素稳定性的因素试验。结果表明:烤后烟叶在贮藏过程中光照强度越强,类胡萝卜素越易降解,而室内避光条件下烤后烟叶类胡萝卜素的稳定性较强;低温贮藏有利于减缓类胡萝卜素的降解,而高温贮藏条件下易加快其分解;干燥的环境有利于维持类胡萝卜素的稳定性,而潮湿环境促进了类胡萝卜素的降解;在Cu2+,Fe2+,Fe3+,Zn2+和Al3+存在条件下类胡萝卜素稳定性较差,但在其他金属离子存在条件下则较稳定;还原剂对类胡萝卜素有一定的保护作用。因此,推荐烟叶在低温、避光、干燥、偏碱性或适宜的还原剂处理下贮藏,通过农业生产措施降低烟叶中Cu2+,Fe2+,Fe3+,Zn2+和Al3+等金属离子含量,有助于抑制烤后烟叶类胡萝卜素降解及烟叶褪色。      

Extraction and quantification of major carotenoids in processed foods and supplements by liquid chromatography

A simple, fast and robust method with minimum steps, small sample size and amounts of solvents was developed to determine major carotenoids contents in processed foods, tablets and gel capsules. The method involves dispersion of the sample in hot water (60 °C) with added butylated hydroxytoluene (BHT) in ethanol to minimize oxidation, followed by extraction with chloroform and analysis by liquid chromatography. Chromatographic parameters were: a C30 column protected with a C18 guard cartridge; gradient elution at the rate of 1.0 mL/min starting with 100% methanol (A) and ending with 40:60 (v/v) methanol/isopropanol (B); detection set at 450 nm for carotenoids, and 325 nm for retinol, retinyl acetate and retinol palmitate. The method exhibited: (i) high degree of repeatability (%rsd); (ii) linear calibration curves (r² ? 0.9998); (iii) low detection; and quantification limits. The method was validated with standard reference material 2383 for trans-β-carotene; and tested for α-, and β-carotenes, lutein, zeaxanthin, cryptoxanthin, trans-retinol in processed foods, tablets and gel supplements.     

Carotenoid profile and retinol content of baby food products

 The contents of provitamin A carotenoids, non-provitamin A carotenoids and retinol were investigated in two types of 65 commercially available baby food products by reverse-phase high-performance liquid chromatography. The content of β-carotene in vegetable products for babies from 5 months was 57% of the total amount of carotenoids (7.83 mg/100 g); the contents of other carotenoids were lycopene 19%, α-carotene 18%, lutein 4%, cryptoxanthin 1% and zeaxanthin 1%. All together, the content of provitamin A carotenoids was 76% and that of non-provitamin A carotenoids was 24%. A similar carotenoid profile was found in vegetable products for babies from 8 months and from 12 months, although the total carotenoid content was lower (3.07 mg/100 g). In fruit and cereal products the contents of β-carotene and lycopene were 36% and 39% of the total amount of carotenoids (0.44 mg/100 g). The contents of α-carotene, lutein, cryptoxanthin and zeaxanthin ranged from 3% to 9% of the total carotenoids. The contents of provitamin A and non-provitamin A carotenoids were equal for this group (both 50% of all carotenoids). The calculated average content of retinol equivalents (mg RE/100 g) was between 0.33 and 0.88 for the vegetable products and 0.03 for the fruit and cereal products. Received: 27 May 1999 / Revised version: 2 August 1999