萝卜硫素和莱菔素提取、纯化及检测方法研究进展

流行病学研究表明,西兰花、萝卜等十字花科蔬菜具有抗炎、抗癌、预防心血管疾病等功能作用,主要是由于这类蔬菜富含硫代葡萄糖苷类化合物,被切碎或咀嚼后水解生成异硫氰酸酯类物质。萝卜硫素和莱菔素是抗癌活性最高的天然异硫氰酸酯类化合物,文中阐明了萝卜属和芸薹属的几种主要蔬菜及种子中萝卜硫素及莱菔素的提取纯化方法,着重概述了常用的萝卜硫素和莱菔素的溶剂萃取法及现代提取技术;比较了1,2-苯二硫醇（BDT）比色法、高效液相色谱法（HPLC）、超高压液相色谱法（UPLC）和气相色谱-质谱联用技术（GC-MS）等萝卜硫素和莱菔素定量分析方法;分析了开口柱色谱层析、固相微萃取（SPE）、高速逆流色谱分析（HSCCC）、制备液相色谱法等纯化方法的优缺点;并对各类提取检测、分离纯化方法进行了分析和总结,讨论了萝卜硫素和莱菔素提取纯化技术的未来发展趋势,以期对萝卜等十字花科蔬菜的加工利用及营养保持技术提供理论参考。     

Optimized Formation,Extraction, and Determination of Sulforaphane in Broccoli by Liquid Chromatography with Diode Array Detection

A new method has been developed to determine sulforaphane in broccoli florets, stems, and leaves using liquid chromatography coupled to diode array detection. The hydrolysis reaction, required to convert glucoraphanin to its breakdown product, sulforaphane, and the extraction procedure, based on solvent extraction with methyl t-butyl ether followed by solid phase extraction with silica cartridges, were optimized for the three broccoli matrices; the resulting recovery values ranged from 92 to 102 % in all cases. A fast separation was performed on a C₁₈ analytical column with a mobile phase composed of ammonium formate (20 mM) in water and acetonitrile (55:45, v/v) in isocratic elution mode. This method was fully validated and was found to be selective, linear from 2.5 to 800 mg/kg, and precise (percent relative standard deviation values below 5 %). Moreover, the limits of detection and quantification were 0.8 and 2.5 mg/kg, respectively. Finally, the proposed method has been applied to the analysis of sulforaphane in broccoli samples of two different cultivars (Parthenon and Marathon).     

高效液相色谱法测定肠衣中三聚氰胺

建立了用酸性氧化铝脱脂、固相萃取净化与高效液相色谱测定肠衣中三聚氰胺的检测方法.试样用三氯乙酸溶液-乙腈超声提取,酸性氧化铝脱脂后经阳离子交换固相萃取柱净化,用高效液相色谱测定,光谱定性、外标法定量.方法的线性范围为1～100mg/L ,相关系数为0.9999,检出限(S/N=10)均达到1mg/kg,在三个浓度水平(1, 5,10mg/kg)进行精密度和回收率试验,所得相对标准偏差在3.3%～4.8%之间,回收率在80%～110%之间.     

Determination of ergosterol in paddy rice using solid phase extraction

Ergosterol, the predominant sterol found in most fungi, is considered as an indicator of fungal invasion in grain. An extraction method based on solid phase extraction (SPE) has been developed for determination of ergosterol in unmilled rice, generally called paddy. The samples obtained by extraction through SPE have been analysed by HPLC. The method has been validated in experiments with paddy subjected to different drying treatments and recoveries above 85% have been obtained. SPE appeared to be simple, quick, reliable and consistent for ergosterol analysis. The ergosterol levels obtained in the paddy samples were found to be within the expected range and were comparable with those found using conventional methods. Copyright © 2004 Society of Chemical Industry     

固相萃取-高效液相色谱法测定畜禽肉中复合硝基酚钠残留

通过碱性溶液提取,再用正己烷萃取除去脂肪,用乙酸调p H为酸性条件后用乙酸乙酯-异丙醇反萃取,浓缩后再通过混合阴离子固相萃取柱净化,最后采用高效液相色谱分离检测。结果:三种复合硝基酚钠组分在0.05¹.0μg/m L范围内其浓度与峰面积呈线性,相关系数均可达到0.999;通过萃取、反萃取以及固相萃取等多种手段净化,样品基质几乎无干扰,检出限可达到0.003⁰.005 mg/kg;通过对0.25、0.5、1.0μg进行加标测试,其回收率在81%⁹5%之间,相对标准偏差均小于10%。      

硬脂酰乳酸钠对铜皂络合比色法测定酸价干扰的固相萃取清除工艺

为清除合成乳化剂硬脂酰乳酸钠(SSL)对铜皂络合比色技术测定油脂酸价过程的正向干扰,开发了一种固相萃取(SPE)清除SSL的技术。用含2%异丙醇的环己烷为固相萃取流动相和油脂样品溶解液,通过离子排斥色谱检测油脂皂化水解后释放的游离乳酸的含量,间接评估SPE净化处理前后油脂中的SSL含量,并结合铜皂络合比色技术测定的酸价变化,评价所建立的SPE技术对去除油脂中SSL的效果。结果表明：SPE技术可有效去除油脂中的SSL,明显消除油脂中SSL对铜皂络合比色法测定酸价的干扰,不会导致油脂样品脂肪酸含量损失。对于含SSL的食用油脂试样,经SPE技术净化处理后,铜皂络合比色法测定酸价过程的干扰明显降低。     

西兰花粉营养成分及理化性质研究

为了促进西兰花茎叶的可食化利用，解决西兰花尾菜极大浪费以及废弃带来的环境问题，本文以西兰花全株为原料，分为可食花球、茎和叶3部分，系统分析了不同部位冻干粉的部分化学成分、矿物质含量、热稳定性及粉体表面形貌；进而对比了不同粒度下粉体的色度、密度、膨胀力和持水持油能力。结果表明，西兰花叶冻干粉中总黄酮含量高达19.82±0.86 mg/g，叶绿素含量为11.35±0.65 mg/g，钙含量高达11.27±0.48 mg/g，铜含量为20.23±0.81 μg/g，锰含量为29.62±0.71 μg/g，上述成分均显著高于花球和茎冻干粉（P<0.05）。相同目数样品的粒径和比表面积具有差异，其中西兰花茎粒径较大，比表面积较小。随着粒径的减小，样品压缩度升高；其中，180目粒度下西兰花叶和茎压缩度较高且差异不显著（P>0.05）。3种样品相比，西兰花叶和花球具有较高的膨胀力，茎和花球具有较高的持水力和持油力。不同样品中，140目花球粉末的持水力、持油力最大，分别为13.18±0.46 g/g和1.77±0.13 g/g。综上所述，综合考虑加工成本，西兰花废弃茎、叶和花球经干燥粉碎粒度选择140目较为适宜。     

An update procedure for an effective and simultaneous extraction of sesquiterpene lactones and phenolics from chicory

A new method for recovering and purifying sesquiterpene lactones (SLs) and optimise the solvent system for their extraction from freeze-dried chicory leaves was studied. Catalogna (leafy chicory) and Head Radicchio types were used as test samples. Solid phase extraction (SPE) employing silica-based cartridges allowed a fast and straightforward purification of SL from interfering phenolics, before HPLC determination. Under these conditions, SL were eluted during loading and washing steps whereas polar phenolic compounds were retained on silica stationary phase. Hydrophilic mixtures methanol/water acidified by small percentages of formic acid led to consistently higher SL recoveries in comparison to organic solvents and enabled the simultaneous extraction of phenolics. Phenolic recovery in crude extracts was assessed at levels higher than 95% for both Catalogna and Head Radicchio. Total, free and bound SL amounts detected in extract obtained by 2% (v/v) formic acid in methanol/water 4/1 (v/v) were 2223.4, 778.5 and 1444.9 mg/kg of dried product in Catalogna, and 401.2, 105.1 and 296.0 in Head Radicchio. In both cases, recoveries were consistently higher than those obtained with methanol alone.     

HPLC测定亚麻籽油苦味肽的纯化富集前处理研究

比较了高效液相色谱(HPLC)定量测定亚麻籽油苦味肽(CLE)的溶剂萃取法和固相萃取法(SPE)纯化富集前处理效果,并对不同规格SPE硅胶小柱填料的前处理效果作了对比。结果表明,通过溶剂萃取法和固相萃取法前处理的亚麻籽油样品,进行HPLC定量测定时,CLE均可以在液相色谱中得到良好的分离,前处理方法并不影响CLE的出峰时间及峰形。但与溶剂萃取法相比,SPE所提取的环肽样品中甘油三酯及其他杂质显著减少,这有利于保护液相色谱柱、提高柱效。随着SPE硅胶小柱填料量的增加,CLE测定值减小,使用填料规格为0. 5 g的SPE硅胶小柱可以得到较好的分离及富集效果。SPE硅胶小柱预处理所得物经超高效液相色谱-质谱联用分析证实为CLE。SPE可有效提取富集亚麻籽油所含的CLE,适用于HPLC定量检测CLE的前处理,该方法样品加标回收率为92. 62%,组内精密度相对标准偏差为0. 62%,组间精密度相对标准偏差为1. 79%。     

SPE - UHPLC - DAD法测定植物油微量酚酸类化合物的方法研究

建立了固相萃取-高效液相色谱对植物油中9种微量酚酸类化合物进行测定的方法。从提取及净化方式、流动相组成、流动相流速以及色谱柱温度等条件优化9种酚酸类化合物的检测方法。结果表明,样品经正己烷溶解,二醇基固相萃取柱(Diol-SPE)净化,甲醇-水为流动相,梯度洗脱,流速0.3 mL/min,检测波长为280 nm时,9种酚酸类化合物的检测结果最好,该检测方法的回收率在91.35%～103.21%之间,日内及日间精密度的RSD范围分别在0.3%～0.9%和0.6%～1.1%之间。该方法快速、准确,具有较好的重复性及稳定性,适合对植物油样品中9种酚酸类化合物的检测,结果为植物油的分类及真假判定提供了参考。     

基于高效液相色谱法对农产品中吡虫啉的提取和净化

基于高效液相色谱法,对不同农产品中吡虫啉的不同提取和净化方法进行比较,建立农产品中吡虫啉农药残留的检测方法。样品经乙腈提取后,采用WondaSep GC-e/PSA固相萃取柱净化,以乙腈-0.1%磷酸为流动相梯度洗脱,XBridge C18色谱柱进行分离,二极管阵列检测器检测,外标法定量。考察不同农产品加水或不加水提取对吡虫啉提取的影响,以及固相萃取和QuEChERS对干扰基质的净化效果。研究表明：不同提取方法和净化方法,对不同农产品影响差异较大。该方法中吡虫啉在0.05～10.0 μg/mL范围内线性关系良好（R2≥0.999）,平均回收率在88.3%～109.1%之间,相对标准偏差不大于3.5%。本研究可为农产品中吡虫啉农药残留的监测提供技术支持。     

The determination of niacin in selected foods by capillary electrophoresis and high performance liquid chromatography: acid extraction

A robust method for the determination of niacin in raw and cooked meat and fish samples using acid extraction to liberate the niacin from the food matrix and capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) as the determinative steps is described. Niacin is liberated from the food matrix by autoclaving in the presence of 0.8 M sulphuric acid at 121°C for 2 h. C18 and cation exchange solid phase extraction (SPE) cartridges were used to isolate and concentrate the niacin before analysis. CE analyses were performed with an uncoated 50 μm extended light path fused silica capillary column and a buffer comprising of 7.5% acetonitrile and 92.5% of a 1:1 mixture of 0.02 M potassium dihydrogen orthophosphate and 0.02 M disodium hydrogen orthophosphate pH 7. Saccharin was used as the internal standard. The levels of niacin determined by CE compared favourably with those determined by HPLC (canned ham CE 3.5 mg/100 g, HPLC 3.3 mg/100 g; raw lamb CE 7.7 mg/100 g, HPLC 7.7 mg/100 g). The average recovery of niacin added to the samples prior to extraction was 103%. The limit of reporting for the determination is 0.5 mg/100 g. The procedure was suitable for only a limited number of other food types (e.g. fruit, vegetables and nuts). Alkali extraction of these foods using saturated calcium hydroxide followed by SPE cleanup and CE determination provided more reliable results. The limit of reporting for this procedure was 0.2 mg/100 g for fruit and 0.5 mg/100 g for vegetables.     

固相萃取-高效液相色谱法测定食品基质中5种不同合成红色着色剂

目的建立一种固相萃取-高效液相色谱法(solidphaseextraction-highperformanceliquid chromatography,SPE-HPLC)测定食品中5种人工合成红色着色剂(新红、苋菜红、胭脂红、诱惑红、赤藓红)的方法。方法比较ProElut PWA-2, Cleanert PWAX和Sep-Pak Plus QMA 3种不同SPE小柱的净化效果,比较Eclipse XDB C_(18)、Venusil XBP C_(18)及Agilent TC-C_(18) 3种色谱柱对5种着色剂的分离情况,按照GB 5009.35-2016《食品中合成着色剂的测定》对食品中5种着色剂进行测定。结果 Pro Elut PWA-2 SPE固相萃取小柱进行样品净化效果最优, Agilent TC-C_(18)色谱柱在10 min内有较好的分离度。加标浓度为0.5～50 mg/kg时,该方法回收率为90.3%～103.9%,相对标准偏差(relative standard deviation, RSD)值为1.18%～7.11%。结论该方法快速、准确,适合用于不同食品基质中5种合成着色剂定性、定量测定。     

Use of postharvest UV-B and UV-C radiation treatments to revalorize broccoli byproducts and edible florets

UV-B radiation (5, 10 and 15 kJ m^− 2) treatments, single or combined with UV-C (9 kJ m^− 2), were hereby firstly studied as bioprocessing tools to enhance bioactive compounds of broccoli byproducts (leaves and stalks) compared to edible florets during storage (72 h/15 °C). Leaves showed similar total phenolic content (TPC) and antioxidant capacity (TAC) than florets, and 2.5/14.5 higher glucoraphanin/glucobrassicin contents than florets. UV increased initial TPC and TAC of leaves/stalks up to 31–97/30–75 and 20–120/170–420%, respectively. UV-B10 + C induced the highest TPC increase (110%) in leaves while UV-B10 and UV-B10 + C led to the highest TPC of stalks after 48 h. UV-B10 + C increased (34%) glucobrassicin levels of leaves while UV-B15 and UV-B15 + C induced the highest glucoraphanin levels (131 and 117 mg kg^− 1) in florets after 72 h. Conclusively, single or combined postharvest UV-B and UV-C treatments can revalorize such agricultural byproducts and also add value to edible broccoli parts.Industrial relevanceThis work demonstrates the high potential of the use of postharvest treatments with UV-B and UV-C, optimized as single or combined treatments, to be used as bioprocessing tools to enhance the bioactive compounds of broccoli byproducts (leaves and stalks) being compared to edible florets. Thought this innovative application of this technology such agricultural wastes may be highly revalorized with a cheap technology leading to high production of health-promoting compounds with such eco-sustainable technology whose benefits can be even reach to the pharmaceutical industry.     

Determination of Hormones Residues in Milk by Gas Chromatography-Mass Spectrometry

The article presents the use of gas chromatography-mass spectrometry (GC-MS) technique in a method for the determination of 18 anabolic hormones from synthetic stilbenes, steroids and resorcylic acid lactones (RALs) groups in raw milk and milk powder. Sample preparation consisted of liquid-liquid extraction with diethyl ether and purification by solid phase extraction (SPE). Prior to instrumental analysis, the reaction of derivatisation with the heptafluorobutyric anhydride or N-methyl-N-trimethylsilyltrifluoroacetamide was performed. Method validation was carried out according to the required performance criteria of the Commission Decision 2002/657/EC. The apparent recovery of all analytes at 1 μg L^?1 (kg^?1) level was ranged between 70.4 and 119.4 % with the coefficients of variation values less than 30 %. The decision limits (CCα) and the detection capabilities (CCβ) were in the range of from 0.11 to 0.44 μg L^?1 (kg^?1) and from 0.19 to 0.75 μg L^?1 (kg^?1), respectively. The procedure has been accredited and successfully applied as a screening method for the presence of hormone residues in the study of commercial samples of milk.     

高效液相色谱-二极管阵列检测器结合固相萃取法快速测定食品中米酵菌酸残留

建立WAX混合型弱阴离子固相萃取富集净化,高效液相色谱-二极管阵列检测器快速测定食品中米酵菌酸残留的方法。采用C18色谱柱（4.6 mm×250 mm,5 μm）,以乙腈-甲醇-1%乙酸溶液（10∶62∶28,V/V）为流动相,以269 nm为分析波长,米酵菌酸在0.1～40 mg/L质量浓度范围内线性良好,相关系数r值大于0.999。本实验考察样品的提取条件及不同固相萃取小柱净化效果,结果表明,选择体积分数80%乙腈溶液为提取溶剂,超声提取30 min,WAX小柱富集、净化,效果最佳。方法检出限为0.03 mg/kg。通过不同样品加标验证,平均回收率为83.3%～95.6%,相对标准偏差不大于5%（n=6）。与GB/T 5009.189-2003《银耳中米酵菌酸的测定》方法相比,该方法精确灵敏、回收率高、前处理简单快速,适用于各种食品中米酵菌酸残留快速测定。     

Solid-phase extraction for HPLC analysis of dietary folates

Phenyl and cyclohexyl bonded silicas used for the solid-phase extraction (SPE) of dietary folates were studied and compared in terms of their retention capacities and recoveries. The SPE procedure was optimised to achieve the maximum recovery. The stabilities of folates in elution buffers and during the SPE procedure were also studied. The use of ascorbic acid throughout the SPE procedure was found to be important for the stabilisation of tetrahydrofolate – the most labile folate compound, whereas the use of both ascorbic acid and 2-mercaptoethanol was necessary to stabilise tetrahydrofolate during the storage of the eluates in the autosampler until analysis. Two purification methods, using phenyl endcapped (PH EC) cartridges or the combination of strong anion-exchange (SAX) and PH EC cartridges, were applied to the determination of folates in different food extracts, and verified by correlation with the usual purification method that uses SAX cartridges. The combination of SAX and PH EC cartridges was found to give the cleanest extracts.     

微波提取-高效液相色谱法测定食用菌中百草枯

建立百草枯在食用菌中的高效液相色谱残留分析方法。样品用10mL浓度2mol/L的盐酸溶液120℃微波消解提取30min,提取液经强阳离子交换(SCX)固相萃取小柱净化,氯化铵饱和溶液洗脱并定容。采用AtlantisHILIC Silica色谱柱,乙腈-辛烷基磺酸钠缓冲液为流动相,紫外检测波长259nm。该方法百草枯定量限为1.0μg/kg(RSN=10)。百草枯的添加回收率为82.33%～96.99%,相对标准偏差为1.23%～3.90%,该方法适合于食用菌中百草枯残留快速分析。     

Evaluation of a solid phase extraction procedure for the determination of pesticide residues in foodstuffs

An evaluation of a method based on solid phase extraction (SPE) as a clean‐up stage for the determination of pesticide residues in composite vegetable and cabbage samples is presented. Samples are extracted with acetonitrile and the extract is loaded onto an SPE cartridge. The pesticides are eluted with a mixture of acetonitrile/toluene (3:1 v/v) and determined by GC–MS or HPLC with fluorescence detection. Acceptable recoveries were obtained for 31 pesticides. An investigation of the effect of partial, as opposed to exact, matrix matching of standards in GC–MS analysis is also presented. While the variation using unmatched standards is visibly greater, the difference is not such as to require exact matrix matching for residue‐screening work. The procedure removes the need for chlorinated solvents, reduces the volumes of other solvents used and allows increased sample throughput compared with liquid–liquid extraction procedures. © 1999 Society of Chemical Industry     

Simultaneous determination of five aluminum lake dyes in chewing gum by HPLC with photodiode array detection

A simple and rapid method has been developed and validated for the determination of five food aluminum lake dyes (Tartrazine Al lake, Sunset Yellow Al lake, Ponceau 4?R Al lake, Allura Red Al lake and Brilliant Blue Al lake) in chewing gum. The dye portions of the target aluminum lakes were simultaneous extracted with 0.25?M NaOH and cleaned up by liquid-liquid extraction with dichloromethane, followed by further purification using Oasis WAX solid-phase extraction (SPE) cartridges. Analytes were separated by HPLC using an Inertsil ® ODS-3 column coupled to a photodiode array detector. The amounts of the aluminum lake dyes were finally quantified and indicated as their dye portions using corresponding calibration curves over ranges of 0.5 to 50?µg?ml^?1, with correlation coefficients >0.9999. Recoveries of the dye parts in aluminum lake dyes (spiked at levels of 1, 5, 25?µg?g^?1) ranged from 72.5 to 116.4%, with relative standard deviations between 0.9 and 6.5%. Limits of detection and limits of quantification for all analytes were 0.15 and 0.50?µg?g^?1, respectively. This method was successfully applied in real samples of chewing gum.