漂浮型和定生型铜藻中色素的液质联用分析

为探究由气候变化、浅海海水污染加剧等因素造成的铜藻爆发性漂浮增殖现象,明确铜藻的漂浮生长机制,采用高效液相色谱-三重四级杆串联质谱(HPLC-QqQ-MS)技术建立11种色素(岩藻黄素、角黄素、玉米黄质、叶黄素、叶绿素a、叶绿素b、α-胡萝卜素、β-胡萝卜素、ε-胡萝卜素、γ-胡萝卜素和 ζ-胡萝卜素)的定性定量分析方法,分析不同地区、不同生态型及不同部位的铜藻中色素的种类和含量。结果表明,共检测到10种色素,其中东极岛漂浮型铜藻中的叶绿素a、叶绿素b和β-胡萝卜素含量显著高于其他地区,而叶黄素、玉米黄质和岩藻黄素含量最低;定生型铜藻的色素总量是漂浮型铜藻的1.6倍;铜藻气囊中含有大量的叶黄素、岩藻黄素、玉米黄质、叶绿素a和叶绿素b等5种色素。本研究建立了快速、灵敏的液相色谱质谱联用定量分析方法,比较分析了不同铜藻中的10种色素种类和含量分布规律,为铜藻的健康、可控利用研究提供了基础数据支撑。     

缺磷胁迫加重柑橘叶片光合作用的光抑制及叶黄素循环的作用

用营养液培养的方法 ,研究了不同光强和磷水平对温州蜜柑叶片光合作用光抑制的影响。结果表明 ,在缺磷强光条件下 ,叶片的初始荧光Fo、最大荧光Fm、PSⅡ光化学效率 (Fv/Fm)及表观电子传递速率 (ETR)下降 ,叶片非光化学猝灭的慢相 (qNs)和叶温升高。用叶黄素循环的抑制剂DTT处理叶片后 ,Fo升高。这些结果说明 ,缺磷胁迫加重了温州蜜柑叶片光合作用的光抑制 ,叶黄素循环的非辐射能量耗散在保护光合机构免受强光破坏方面起着重要作用。     

不同施肥条件对谷子β-胡萝卜素和叶黄素含量的影响

为了探究不同施肥条件对谷子类胡萝卜素含量的影响及筛选谷子高类胡萝卜素含量的施肥种植条件,运用L16(45)正交试验,以16种不同氮、磷、钾施肥配比作为长农35号谷子的施肥种植条件,测定各施肥条件下谷子β-胡萝卜素和叶黄素含量,并对其进行分析。结果表明,不同施肥条件对谷子β-胡萝卜素和叶黄素含量均有显著影响,最佳施肥条件为尿素485 g、重过磷酸钙0 g、硫酸钾300 g,此条件下谷子的β-胡萝卜素和叶黄素总含量最高。通过方差分析和偏相关分析得到氮、磷、钾肥对谷子β-胡萝卜素和叶黄素含量的影响大小依次为氮肥钾肥磷肥。本研究为培育高类胡萝卜素含量谷子所需的施肥条件提供了理论依据和现实基础。     

酶为介质有机溶剂提取万寿菊花中叶黄素的工艺研究

以万寿菊(Tagetes erecta)花粉末为原料，采用液态纤维素酶处理和有机溶剂萃取同时进行的方法提取叶黄素，研究并优化此过程的工艺条件。试验结果表明：酶为介质有机溶剂提取万寿菊花中叶黄素工艺条件受多种因素影响，其中提取时间、萃取前保温时间、酶浓度、搅拌速度对提取过程影响较大。确定酶法提取较适宜的工艺条件为：2.000 g粒度60目的万寿菊花粉末、液态纤维素酶浓度为7%(m/m)、液料比为20∶1、萃取前保温时间为1.5 h、搅拌速度为700 r/min、提取时间为3 h，对此工艺条件进行验证叶黄素提取率可达92.37%。而单纯的有机溶剂法叶黄素提取率仅为77.53%，此方法明显优于单纯的有机溶剂法。     

超临界CO2萃取万寿菊花中叶黄素的研究

采用超临界CO2萃取技术,研究了从万寿菊花中萃取叶黄素的工艺条件.对影响超临界CO2萃取叶黄素的各种因素,包括分离参数、原料含水率、粉碎粒径,超临界萃取温度、压力、流速、时间等因素进行了考察,得到较佳的萃取工艺条件为:原料含水率10.92%,粒径40目,萃取温度60℃,压力30 MPa,CO2流速15 L/h,分离釜Ⅰ温度40℃,压力6 MPa,分离釜Ⅱ温度20℃,时间为6 h.     

超临界CO2萃取万寿菊花中叶黄素的研究

采用超临界CO₂萃取技术,研究了从万寿菊花中萃取叶黄素的工艺条件。对影响超临界CO₂萃取叶黄素的各种因素,包括分离参数、原料含水率、粉碎粒径，超临界萃取温度、压力、流速、时间等因素进行了考察,得到较佳的萃取工艺条件为:原料含水率10.92％，粒径40目，萃取温度60℃，压力30 MPa，CO₂流速15 L/h，分离釜Ⅰ温度40℃，压力6 MPa，分离釜Ⅱ温度20℃，时间为6 h。     

小米脂氧合酶活性与储藏稳定性关系

为研究小米中脂氧合酶(LOX)活性和储藏稳定性的关系,初步探索小米褪色机制,以褪色快的小米品种(‘谷丰2号’、‘公谷63号’)、褪色慢的小米品种(‘大青秸’、‘红苗牛头沟’)为试验材料,对其进行人工老化处理,并测定人工老化处理前后小米的脂氧合酶活性、米色、类胡萝卜素、丙二醛(MDA)含量的变化。同时,利用HPLC对小米类胡萝卜素提取液进行分析,比较4个小米品种中叶黄素、玉米黄质及β-胡萝卜素的变化差异,分析小米脂氧合酶活性与类胡萝卜素及其主要组分的相关性。结果表明,经老化处理后,4个小米品种的LOX活性、米色b*值和类胡萝卜素含量均下降,MDA含量升高。‘大青秸’、‘红苗牛头沟’小米与‘谷丰2号’、‘公谷63号’小米相比,其米色、类胡萝卜素含量和MDA含量变化较小,储藏稳定性较好。HPLC分析表明,老化处理前后4个小米品种叶黄素、玉米黄质的变化无明显差异,而β-胡萝卜素含量的变化存在明显差异;相关性分析表明,老化处理前后LOX活性与β-胡萝卜素含量呈负相关,表明LOX可能与β-胡萝卜素发生偶联,进而导致小米类胡萝卜素含量降低。本研究结果为耐储藏小米的遗传育种提供了一定的理论基础。     

盐生杜氏藻β-胡萝卜素羟化酶基因(chyb)的克隆及表达分析

玉米黄质(3,3'-二羟基-β-胡萝卜素)是自然界中常见的一种β-胡萝卜素衍生物,是黄斑和视网膜中最关键的部分.玉米黄质是通过一系列酶作用合成,其中β-胡萝卜素羟化酶(chyb)是关键限速酶,为了研究盐生杜氏藻(Dunaliella salina)中不同胁迫下β-胡萝卜素羟化酶基因的表达及玉米黄质含量的变化.本研究采用RACE方法首次从盐生杜氏藻中扩增出β-胡萝卜素羟化酶基因(Dschyb),并分析强光、葡萄糖等因素对DSchyb表达及玉米黄质含量的影响.结果表明,该基因全长1 433 bp(GenBank登录号:JN118489),包含一个969bp的开放阅读框,编码322个氨基酸,氨基酸的分子量为35.51kD,等电点(PI)为9.01.DsCHYB包含有4个保守的组氨酸基序,与团藻及莱茵衣藻的同源性分别是64％和58％.DsCHYB具有4个跨膜结构及叶绿体导肽,进一步证明该酶定位于叶绿体类囊膜上.系统进化树分析标明,DsCHYB与其他绿藻如团藻(Volvox carteri f.Nagariensis)、莱茵衣藻(Chlamydomonas reinhardtii)的CHYB共处一个进化支,亲缘关系很近.Dschyb基因的表达调控研究显示,在经强光刺激24 h后,Dschyb 基因表达显著上调(P＜0.01),在48 h表达最高(P＜0.01).在经乙酸钠、硫酸亚铁和强光共同处理6h时,Dschyb表达急剧上升(P＜0.01),处理12h后下降;在葡萄糖处理1.5h后,其表达达到最高,6h后表达与对照组无显著性差异;添加放线菌素D后,Dschyb的表达相对对照降低,表明放线菌素D可能对葡萄糖诱导Dschyb表达具有抑制作用.高效液相色谱测定其玉米黄质含量,钠铁、强光及葡萄糖处理均能提高盐生杜氏藻的玉米黄质的含量,其玉米黄质含量比对照组分别增长16％(P＜0.05),28％(P＜0.05)和53％(P＜0.01).本研究结果为玉米黄质合成调控及其在藻类中功能研究提供理论指导.     

检测睾酮、孕酮、雌酮与雌三醇的双抗体酶免疫分析法

本文报道了自制的试剂建立了检测睾酮、孕酮、雌酮和雌三醇的双抗体酶免疫分析法。实验表明 ,这些测定具有很高灵敏度 (最小可测量为 0 1～ 1pg 孔 ) ,标准曲线范围为 0～ 40pg 孔 ;同相关类固醇激素 (孕酮、孕烯醇酮、睾酮、雄酮、雌酮、雌三醇和皮质酮等 )的交叉反应率一般在 0 1 %以下。测定的批内和批间变异系数分别在5 8%～ 7 2 %与 9 5%～ 1 0 5% (n =8)范围内。向血中添加激素标准品后的回收率在 94%～ 1 1 0 %范围内。用自制防腐剂处理后 ,液态抗体 (包括第二抗体 )、酶标记物和已包被第二抗体的滴定板可保持活性 5～ 1 2个月或更久 ( 4℃～ -2 0℃ )。此测定可节约激素特异抗体 4～ 1 0倍。初测人和动物的血、奶、尿样品表明所建立的 4种测定方法可分别用于人和动物体液中睾酮、孕酮、雌酮和雌三醇的定量检测     

Cu、Zn、Pb、Cd对鲫鱼(Carassius auratus)组织DNA毒性的研究

用非程序DNA的合成作为毒性评价指标研究了混合重金属对鲫鱼 (carassiusauratus)组织DNA的毒性作用 ,结果表明 ,混合重金属的毒性作用靶标器官是鱼脑和鱼的肝脏 ;混合重金属中的主要毒性成分是Zn、Pb和Cd离子 ,混合重金属对鱼脑组织的毒性大小顺序是Pb>Cd >Zn ,对肝脏DNA的毒性顺序为Pb >Zn >Cd;Cu离子能够降低其它 3种离子的毒性作用 ,降低的机制在于竞争作用 ,导致了其它离子的吸收降低     

Thermal processing of vegetables increases cis isomers of lutein and zeaxanthin

Carotenoids, lutein and zeaxanthin, found in fruits and vegetables, comprise the macula pigment of the eye. These carotenoids exist in plants as the all-trans geometric form; however, in human plasma, cis isomers of these carotenoids have also been identified. Thermal processing can induce carotenoid trans to cis isomerization. The aim of this research was to determine if thermal processing induces isomerization of lutein and zeaxanthin and to quantify the extent of this reaction. High-performance liquid chromatography was used to separate and quantitate geometric isomers of lutein and zeaxanthin. Isomers were tentatively identified by UV-visible absorbance spectra, comparison of retention times to those of isomerized standards using C(30) chromatography, and mass spectrometry. Thermal processing increased the percent cis isomers of lutein and zeaxanthin up to 22 and 17%, respectively. Further studies are needed to consider the physiological impact of consuming carotenoid isomers in processed vegetables.     

Distribution of lutein,zeaxanthin, and related geometrical isomers in fruit,vegetables, wheat,and pasta products

Quantitative data with regard to dietary (3R,3'R,6'R)-lutein, (3R,3'R)-zeaxanthin, and their (E/Z)-geometrical isomers are scarce, and in most cases, only the combined concentrations of these two carotenoids in foods are reported. Lutein and zeaxanthin accumulate in the human macula and have been implicated in the prevention of age-related macular degeneration (AMD). The qualitative and quantitative distributions of lutein, zeaxanthin, and their (E/Z)-isomers in the extracts from some of the most commonly consumed fruits, vegetables, and pasta products were determined by HPLC employing a silica-based nitrile-bonded column. Green vegetables had the highest concentration of lutein (L) and zeaxanthin (Z), and the ratios of these carotenoids (L/Z) were in the range 12-63. The yellow-orange fruits and vegetables, with the exception of squash (butternut variety), had much lower levels of lutein in comparison to greens but contained a higher concentration of zeaxanthin. The ratio of lutein to zeaxanthin (L/Z) in two North American bread varieties of wheat (Pioneer, Catoctin) was 11 and 7.6, respectively, while in a green-harvested wheat (Freekeh) imported from Australia, the ratio was 2.5. Between the two pasta products examined, lasagne and egg noodles, the latter had a much higher concentration of lutein and zeaxanthin. The levels of the (E/Z)-geometrical isomers of lutein and zeaxanthin in these foods were also determined.     

Xanthophylls in commercial egg yolks: quantification and identification by HPLC and LC-(APCI)MS using a C30 phase

The xanthophylls lutein and zeaxanthin have attracted a lot of interest since it was presumed that an increased nutritional uptake may prevent adult macula degeneration (AMD). Although egg yolks serve as an important dietary source of lutein and zeaxanthin, data on xanthophyll concentrations in commercial egg yolks are not available. Thus, an high-performance liquid chromatography-diode array detector (HPLC-DAD) method was developed allowing for simultaneous separation of eight xanthophylls used to fortify poultry feed. Peak identification was carried out by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [LC-(APCI)MS]. Egg yolks of four types of husbandry (seven batches each) were examined. Lutein and zeaxanthin were the predominant xanthophylls in egg yolks produced in accordance with ecological husbandry (class 0) because the concentrations of these xanthophylls ranged from 1274 to 2478 microg/100 g and from 775 to 1288 microg/100 g, respectively. Analysis of variance (ANOVA) proved that both mean lutein and mean zeaxanthin concentrations of eggs from class 0 were statistically discriminable from mean lutein and zeaxanthin concentrations from eggs of all other classes (P < 0.01). Mean concentrations of synthetic xanthophylls in eggs of classes 1 (free range), 2 (barn), and 3 (cage) were as follows: canthaxanthin, 707 +/- 284 microg/100 g; beta-apo-8'-carotenoic acid ethyl ester, 639 +/- 391 microg/100 g; and citranaxanthin, 560 +/- 231 microg/100 g. Experiments with boiled eggs proved that beta-apo-8'-carotenoic acid ethyl ester was the xanthophyll with the highest stability, whereas lutein was degraded to the largest extent (loss of 19%). Detailed knowledge about the xanthophyll amounts in eggs is indispensable to calculate the human uptake.     

Quantification of carotenoid and tocopherol antioxidants in Zea mays.

Recent investigations into carotenoid and tocopherol biological activity in mammalian systems indicate that these antioxidants are associated with the prevention of degenerative diseases. Both carotenoids and tocopherols can be found in corn kernel tissue. A replicated survey of 44 sweet and dent corn lines was conducted to determine qualitative and quantitative variability of lutein, zeaxanthin, beta-cryptoxanthin, alpha-carotene, and beta-carotene, as well as the alpha-, delta-, and gamma- forms of tocopherol. The primary carotenoids in fresh market sweet corn were found to be lutein and zeaxanthin, with the gamma form dominating among the tocopherols. Mean values among the genotypes were observed to range from 0 to 20.0 and 2.4 to 63.3 microg/g dry weight for lutein and gamma-tocopherol, respectively, indicating variability among genotypes in genes regulating the metabolism of these compounds. The observed genetic variability suggests profound differences in potential health promotion among genotypes and supports the feasibility of developing germplasm with enhanced levels of these antioxidant compounds at dosages that could promote health among the consuming public.     

Improved normal-phase high-performance liquid chromatography procedure for the determination of carotenoids in cereals

Besides the health benefits associated with whole-grain consumption, cereals are recognized sources of health-enhancing bioactive components such as carotenoids, which are a group of yellow pigments involved in the prevention of many degenerative diseases and which have been used for a long time as indicators of the color quality of durum wheat and pasta products. This work reports a fast, sensitive, and selective procedure for the extraction and determination of carotenoids from cereals and cereal byproducts. The method involves sample saponification and extraction followed by normal-phase high-performance liquid chromatography, allowing the separation of the main carotenoids pigments of cereals, especially lutein and zeaxanthin. An application of the established method to various species of cereals and cereal byproducts is also shown. The highest carotenoid levels were found in maize (approximately 11.14 mg/kg of dry weight), which contains high amounts of beta-cryptoxanthin (2.40 mg/kg of dry weight), and, among the cereals considered, has the highest content of zeaxanthin (6.43 mg/kg of dry weight) and alpha+beta-carotene (1.44 mg/kg of dry weight). With the exception of maize, lutein is the main compound found (from 0.23 to 2.65 mg/kg of dry weight in oat and durum wheat, respectively). Moreover, whereas alpha+beta-carotene and zeaxanthin are principally localized in the germ, lutein is equally distributed along the kernel.     

Lutein and zeaxanthin in leafy greens and their bioavailability: olive oil influences the absorption of dietary lutein and its accumulation in adult rats

This study determined the lutein level in various green leafy vegetables (GLVs) and the influence of olive and sunflower oils on the postprandial plasma and eye response of dietary lutein in adult rats, previously induced with lutein depletion (LD). Fresh GLVs (n = 35) were assessed for lutein (L) and its isomer zeaxanthin (Z) levels by high-performance liquid chromatography and liquid chromatography-mass spectrometry. Among GLVs analyzed, Commelina benghalensis L. contained a higher level of L + Z (183 mg/100 g dry wt) and was used as a lutein source for feeding studies. Rats with LD were fed a diet containing powdered C. benghalensis (2.69 mg lutein/kg diet) with either olive oil (OO group), sunflower oil (SFO group), or groundnut oil (GNO group) for 16 days. The L + Z levels of the OO group were markedly (p > 0.05) higher than those of SFO and GNO groups, in plasma (37.6 and 40.9%) and eyes (22.7 and 30.8%), respectively. These results suggest that oleic acid or OO can be used as a suitable fat source to modulate the absorption of dietary lutein to manage age-related macular degeneration.     

Carotenoid Composition and Changes in Sweet and Field Corn (Zea mays) During Kernel Development

Kernels of two carotenoid‐rich cultivars, sweet corn Jingtian 5 and field corn Suyu 29, were compared in terms of carotenoid composition during corn kernel development. The results showed that eight principal carotenoids were characterized by HPLC with diode array detection and atmospheric pressure chemical ionization tandem mass spectrometry with a C30 column. During kernel development, there was a similar trend in the change of total carotenoids for both corn cultivars, and the variation of individual carotenoids was also somewhat similar; violaxanthin, zeaxanthin, lutein, α‐cryptoxanthin, and β‐cryptoxanthin contents had upward trends, whereas neoxanthin content declined all the time, and α‐carotene and β‐carotene had no significant changes. However, the highest levels of the major carotenoids lutein (41.61 µg/g, dry weight) and zeaxanthin (39.59 µg/g, dry weight) obtained in field corn Suyu 29 during the milk stage were higher than those in sweet corn Jingtian 5, whereas the other individual carotenoid levels were significantly lower. Compared with the grain color, highly significant positive correlations were observed between zeaxanthin, lutein, and violaxanthin contents and deeper yellow/orange coloration indicators for field corn Suyu 29, but these relationships were weak for sweet corn Jingtian 5. Potential genetic variation might exist for carotenoid accumulation in sweet and field corn kernels.     

Identification and quantification of zeaxanthin esters in plants using liquid chromatography-mass spectrometry

It has been suggested that lutein and zeaxanthin may decrease the risk for age-related macular degeneration. Surprisingly, oleoresins rich in zeaxanthin are not yet available on the market. Several authors have reported enhanced stability of esterified xanthophylls, so plants containing zeaxanthin esters were investigated to establish valuable sources for the production of durable oleoresins. Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [LC-(APCI)MS] was used to unequivocally identify zeaxanthin esters of a standard mixture and in several plant extracts. Zeaxanthin esters were quantified on the basis of their respective molecular masses using zeaxanthin for calibration; total zeaxanthin was determined after saponification of aliquots of the extracts. Thus, dried wolfberries (Lycium barbarum), Chinese lanterns (Physalis alkekengi), orange pepper (Capsicum annuum), and sea buckthorn (Hippophae rhamnoides) proved to be valuable zeaxanthin ester sources. The present LC-MS method allows for an even more detailed analysis of zeaxanthin esters than reported previously.     

Analysis of xanthophylls in corn by HPLC

An HPLC method was developed using the C-30 carotenoid column to separate and identify the major xanthophylls in corn (lutein, zeaxanthin, and beta-cryptoxanthin). A photodiode array detector and a mobile phase consisting of methyl tert-butyl ether/methanol/water was used. All three xanthophylls eluted in less than 25 min. Yellow dent corn had a total xanthophyll content of 21.97 microg/g with lutein content of 15.7 microg/g, zeaxanthin content of 5.7 microg/g, and beta-cryptoxanthin of 0.57 microg/g. Commercial corn gluten meal had a 7 times higher concentration of xanthophylls (145 microg/g), and deoiled corn contained 18 microg/g, indicating that the xanthophylls are probably bound to the zein fraction of corn proteins.