HPLC同时测定甘薯茎叶中叶黄素和β–胡萝卜素

以水果薯、水果花叶薯、紫羽1号、紫羽2号、农大紫1号、农大紫2号、浙紫1号、紫薯2号、高淀粉、叶用薯1号、叶用薯2号等11种不同品种甘薯茎叶为试验材料,建立了高效液相色谱法同时测定甘薯茎叶中叶黄素和β–胡萝卜素含量的方法。结果表明：采用甲醇为提取溶剂,料液比1∶30,50℃超声提取3次,每次40 min,能较完全提取甘薯茎叶中的叶黄素和β–胡萝卜素;HPLC以甲醇–水（95∶5）、甲醇–乙腈–四氢呋喃（70∶20∶5）为流动相梯度洗脱,叶黄素和β–胡萝卜素分离良好,回收率分别为105.8%和101.5%;叶黄素及β–胡萝卜素含量最高的是甘薯茎叶中的成熟叶,其次为新叶,茎的含量最低;成熟叶中叶黄素与β–胡萝卜素的含量分别在0.504~1.031 mg/g和0.188~0.467 mg/g,新叶中叶黄素与β–胡萝卜素的含量分别在0.409~0.617 mg/g和0.141~0.242mg/g,叶柄中叶黄素与β–胡萝卜素的含量分别在0.094~0.195 mg/g和0.040~0.082 mg/g,茎中叶黄素与β–胡萝卜素的含量分别在0.053~0.103 mg/g和0.036~0.045 mg/g。

壳聚糖-菜籽油脂体负载β-胡萝卜素体系构建与研究

β-胡萝卜素是人体不可缺少一种天然色素,具有抗氧化、抗癌、抗心血管疾病等功能活性,但极性较低且极易氧化,这使得其在食品体系中的应用与消化吸收具有挑战性,利用乳液体系将其包埋是解决上述问题的有效方法。油脂体作为植物种籽细胞中贮藏甘油三酯的特殊细胞器,具有“单层磷脂嵌套蛋白-油脂”的“膜-核”结构,是天然存在的乳化后的油滴,适宜作为水包油乳液的分散相。本研究采用微热搅拌和层层自组装相结合的方法,利用天然油脂体负电与壳聚糖正电性,将β-胡萝卜素包裹在壳聚糖-菜籽油脂体乳液中,并实现了对其稳定性、包埋性的提高,产生了较小粒径的高电荷阳离子乳液(d3,2=2.25μm;ζ=+66.67 mV)。类胡萝卜素的包封率为81.98%,载药量为1.23 mg/g油脂体;本研究结果为开发纯天然乳液递送体系的理论与应用创新提供了参考。

衣藻β–胡萝卜素加酮酶基因在南瓜果实中的瞬时表达

为了研究β–胡萝卜素加酮酶基因在南瓜中合成酮基类胡萝卜素的功能,采用农杆菌介导瞬时表达技术,将含有衣藻β–胡萝卜素加酮酶基因CrBKT的pBI121-CMTPCRBKT载体和pBI121对照载体的农杆菌菌液注入授粉后2、5、10、15和25 d的南瓜果实中,3 d后观察发现,授粉后2和5 d的果实转化后果肉呈微红,其中授粉后5 d的颜色较深,而10 d以上的无红色色素积累。经高效液相色谱(HPLC)分析色素成分,发现积累的红色色素为酮基类胡萝卜素角黄素和虾青素。与对照相比,注入CrBKT基因的授粉后2和5 d的幼果中的类胡萝卜素含量显著增加,5 d的幼果约增加了1/3,且含有106.31μg·g^(-1)酮类胡萝卜素,其中角黄素占80.65%,虾青素占19.35%,而授粉后10 d以上的果实类胡萝卜素含量没有明显变化,也没有酮类胡萝卜素的积累。PCR扩增表明转化组织中表达了该基因。结果表明,CrBKT基因只能在授粉后5 d以内的幼果中表达,并将幼果中的类胡萝卜素转化为酮基类胡萝卜素;随着果实成熟,菌液在果肉组织中无法渗透而阻碍基因表达。

红色和橙黄色果肉西瓜番茄红素和β–胡萝卜素代谢基因的表达分析

以橙黄色果肉西瓜‘WM-Clr-2’及红色果肉西瓜‘PI 179881’为试材,采用HPLC法分析其番茄红素及β–胡萝卜素在7个发育时期的积累情况,同时采用q RT-PCR技术分析番茄红素和β–胡萝卜素代谢途径中8个关键酶基因(PSY1、PSY2、PDS、ZDS、CRTISO、LCYB、NCED1和NCED7)的表达量。结果显示,随着果实的发育,番茄红素和β–胡萝卜素含量均不断增加,且在同一时期,红色果肉‘PI 179881’中的含量始终大于‘WM-Clr-2’。q RT-PCR结果显示PSY1在‘PI 179881’中的表达量高于‘WM-Clr-2’,LCYB和NCED1在‘WM-Clr-2’中的表达量高于红色果肉‘PI 179881’。分析番茄红素及β–胡萝卜素的积累规律与相关基因表达规律说明PSY1的大幅上调表达,LCYB的下调表达及NCED1的低表达可能是红色果肉‘PI 179881’中番茄红素和β–胡萝卜素大量积累的主要原因。PSY1的低表达水平,LCYB和NCED1的高表达水平可能是橙黄色果肉‘WM-Clr-2’中番茄红素及β–胡萝卜素积累量少的主要原因。

Molecular Cloning and Expression of Carotenogenic Genes in Yellowish and Mutant Whitish Loquat (Eriobotrya japonica) Fruits

ObjectiveThe study aimed to explore the factors regulating carotenoid accumulation in flesh color. MethodA loquat mutation （red-or orange-fleshed plant emerged a bud mutation of white-flesh in trunk） was used as material; HPLC analysis of β-carotene content was conducted. ResultThe β-carotene concentration in the flesh of wild and mutant types was 60.9 and 4.6 μg/g fresh weight, respectively. According to the conserved regions of genes from rose family genome, carotenogenic gene fragments in wild and mutant types were obtained. No nucleotide variation of the carotenogenic gene fragments was observed between wild and mutant genome. Real-time quantitative polymerase chain reaction （Q-PCR） was compared and one carotenogenic gene, β-ring hydroxylase （HYB） were considerably suppressed in mature mutant loquat fruits compared with that in wild. The other six carotenogenic genes were also expressed but the expression patterns appeared to be not correlated with the amount of β-carotene concentration in wild loquat flesh. ConclusionThe mutant whitish loquat lacks the ability to synthesize β-carotene because of the transcriptional down-regulation of carotenogenic gene HYB.

Resonance Raman Spectra of Purified PS Ⅱ Core Antenna Complexes CP43 and CP47

PSⅡ core antenna complexes, CP43 and CP47, were purified from spinach (Spinacia oleracea L.) by DEAE Fractogel TSK 650S anion exchange chromatography. Their normal temperature (298 K) resonance Raman spectra were measured. The results suggest that all β carotenoids bound to CP43 and CP47 are in all trans configuration and likely in twisted conformations.

Dynamic Accumulation of β-carotene and Dry Matter in Orange-fleshed Sweetpotato [Ipomoea batatas(L.) Lam] and Their Correlation Analysis

Using three orange-fleshed sweetpotato varieties as experimental materials,16 economic traits at 60 d,75 d,90 d,105 d,120 d and 135 d after planting were measured,so as to revealing the dynamic variations of β-carotene and dry matter accumulation in roots and their relationships with economic traits in orangefleshed sweetpotato.The results showed that the dynamic variations of β-carotene accumulation in tubers varied hugely among different varieties.Interesting,the βcarotene content of all three varieties showed a significant decrease after 120 d,while the dry matter content of them performed a similar ＂fluctuation-type＂.Correlation analysis indicated that β-carotene content of three orange-fleshed sweetpotato varieties had no significant correlation with dry matter content and photosynthetic parameters,but the correlation with other economic traits also varied among varieties.

Observation of Fundamental and High Order Raman Modes of β-Carotenes in Pd OECC Thin Film by SERS

Raman spectra of purified oxygen evolution core complexes (Pd OECC) thin films on silver mirror substrates have been taken over the frequency range of 250-3100 cm -1 by surface enhanced Raman scattering (SERS). Besides the fundamental frequency modes of β_carotene in Pd OECC, many weak peaks are observed. According to the selection rules of overtone and combination bands, most of them are attributed to the second_order Raman spectra of β_carotene. Compared with the SERS of normal Pd OECC, the SERS of Pd OECC after strong illumination shows a decrease in scattering intensity and an increase in line widths, indicating changes of conformation and micro_environment of β_carotene. The results of SERS are consistent with the changes of absorption spectrum of Pd OECC induced by strong illumination. There are no changes that can be ascribed to new vibration bands, so it is deduced that Pd OECC on the silver mirror is identical to that in the solution. In summary, SERS proved a good method to study the photodamage mechanism of photosynthesis.

Adsorption Kinetics of β-Carotene and Chlorophyll onto Acid-activated Bentonite in Model Oil

The textural properties of acid-activated bentonite （AAB）, which were prepared using four different concentrations of sulfuric acid, were analyzed by adsorption-desorption isotherm of nitrogen using an automated specific surface area and porosity analyzer. The total pore volume, specific surface area and average pore diameter of these four kinds of AAB show a regular changing trend, increasing first and then decreasing, the optimum of which can be achieved at a sulfuric acid concentration of 25% （sample A25）. The kinetic analysis of the adsorption of β-carotene and chlorophyll in model oil solutions, namely, xylene and edible oil solution, has been investigated by using AAB. Experimental results indicated that the adsorption data fit the pseudo-second-order model well. The whole adsorption process of the two pigments on AAB was divided basically into two parts： the initial adsorption of pigments was rapid in the first l0 rain, followed by a slower adsorption process till equilibrium was attained at 60 rain. In addition, the amount and rate of adsorption on A25 increase synchronously with the initial pigment concentration and temperature. The results showed that the adsorption kinetics behavior of AAB with respect to the pigments is not influence by the xylene and edible oil solution.

Application of derivative ratio spectrophotometry for determination of β-carotene and astaxanthin from Phaffia rhodozyma extract

A derivative ratio spectrophotometric method was used for the simultaneous determination of β-carotene and astaxanthin produced from Phaffia rhodozyma. Absorbencies of a series of the standard carotenoids in the range of 441 nm to 490 nm demonstrated that their absorptive spectra accorded with Beer’s law and that the additivity when the concentrations of β-carotene and astaxanthin and their mixture were within the range of 0 to 5 μg/ml, 0 to 6 μg/ml, and 0 to 6 μg/ml, respectively. When the wavelength interval (?λ) at 2 nm was selected to calculate the first derivative ratio spectra values, the first derivative amplitudes at 461 nm and 466 nm were suitable for quantitatively determining β-carotene and astaxanthin, respectively. Effect of divisor on derivative ratio spectra could be neglected; any concentration used as divisor in range of 1.0 to 4.0 μg/ml is ideal for calculating the derivative ratio spectra values of the two carotenoids. Calibration graphs were established for β-carotene within 0?6.0 μg/ml and for astaxanthin within 0?5.0 μg/ml with their corresponding regressive equations in: y=?0.0082x?0.0002 and y=0.0146x?0.0006, respectively. R-square values in excess of 0.999 indicated the good linearity of the calibration graphs. Sample recovery rates were found satisfactory (>99%) with relative standard deviations (RSD) of less than 5%. This method was suc- cessfully applied to simultaneous determination of β-carotene and astaxanthin in the laboratory-prepared mixtures and the extract from the Phaffia rhodozyma culture.

Sand Storage, Extending the Shelf-Life of Fresh Sweetpotato Roots for Home Consumption and Market Sales

Uni-modal rainfall pattern has long dry spell wherein sweetpotato is scarce, expensive but cheap at harvest. The crop is mostly consumed, processed or sold. Extending shelf-life of roots is crucial for Malawi and Northern Ghana as the crop has high value. Trials were conducted in the countries at the community level. In the dry season, temperature is cool in Malawi while warm in Ghana, but thru harmattan, the night is cool with low relative humidity. In Malawi, orange-fleshed sweetpotato Zondeni var., white and yellow types were assessed in three types of storage, Afghan ventilated pit store, storage in dry sand of pit-steps, and of a granary In Ghana, local moistened heap and sandbox were compared. In Malawi, weight losses were calculated relative to the quantity stored at start, it was not cumulative. At 1.5 months no significant difference was among treatments. By 3.5 months the pit-steps method emerged to be superior and continued to 6.5 months. Losses in granary were due to shriveling, in the pit-stepsdue to termites and rats, and in ventilated pit due to termites, rats and Java black rot. Sprouting was high in pit-steps, but it was simply removed and roots returned to storage. At 6.5 months, the beta-carotene of Zondeni roots was traceable. Farmers gained high price when selling them as roots were scarce. Women favored the pit-steps because it was manageable. In Ghana, the sandbox was superior to local moistened heap. Methods designed were suitable for home consumption, but will require modification for commercialization.

Quick Fluorescence Method for the Distinguishing of Vegetable Oils

This paper presents the possibilities offered by fluorescence spectroscopy for the identification of vegetable oils such as soybean, sunflower, flax, walnut, corn, almond, sesame, olive and pumpkin oils. The probes under study have been excited with two types of sources： a laser diode （LD） and light-emitting diodes （LEDs） emitting in the UV and in the visible range. Total luminescence spectra were recorded by measuring the emission spectra in the range 350-720 nm at excitation wavelengths from 375 to 450 nm. The excitation-emission matrices have been obtained and two basic fluorescence regions in the visible have been outlined. On this basis the fluorescence spectra of the oils have been subdivided into three categories depending on the prevalence of the fluorescence maxima. The samples show differences in their fluorescence spectra. The latter fact shows that fluorescence spectroscopy can be used for the quick identification of edible oils. The fatty acid, the tocopherol, the beta-carotene and chlorophyll contents in the analyzed oils have been studied. It is shown that some of the types of oils differ significantly from each other by the first derivatives of their fluorescence spectra. There also exist color differences between the groups of vegetable oils under study.

Effect of Steaming, Steam-Drying and Frying on the Provitamins A and Ascorbic Acid Contents of Squash （ Cucurbita Spp.）

Vitamin A deficiency still remains a nutritional concern in Cameroon. Squashes pulp is rich in provitamins A and could help to reduce this deficiency. Unfortunately, in Cameroon, squash pulp is more often reserved for the nutrition of sick people and domestic animals. This study aimed at encouraging the consumption of squash pulp to contribute to vitamin A needs. The contents of a-carotene, all-trans-β-carotene and ascorbic acid were determined respectively by HPLC and titration with 2.6 dichlorophenol-indophenol in five landraces of raw, steamed, steam-dried and fried squash pulp from Cameroon. Moisture and total lipid content were also determined. Peeled pulp squash of 5 cm slice was steamed at 90℃ for 30 min or at 85℃ for 30 min, sliced again at 5 mm thickness and dried at 80℃ during 4 h to obtained steamed or steam-dried squashes. To have fried squashes, 40 g of 1 mm thickness slices pulp were fried in boiling refined palm oil （free of carotenoids） bath at 150℃ during 7 and 10 min. The results obtained showed that steam-drying and frying of squashes leaded to water losses （89%-95%）. As a consequence of this, the a-carotene, all-trans-β-carotene and ascorbic acid contents （expressed in g （100 g）-1 fresh portion） of steam-dried and fried squashes were significantly higher （P 〈 0.05） than those of steamed and raw pulp. However, the retention rate of provitamins A and ascorbic acid was more elevated in steamed than in steam-dried and fried squashes. These results suggest that steam-dried and fried squashes could contribute to fight against vitamin A deficiency while increasing availability of squashes.

The Protective Role of β-Carotene and Hesperidin on Some Hematological and Myocardial Measurements against Imidacloprid Toxicity in Albino Rats

Neonicotinoids including IM （Imidacloprid） are widely used as plant systemic insecticides. Several studies have indicated that pesticide toxicity may be associated with the enhanced production of ROS （reactive oxygen species）. Both β-carotene （I3C） and hesperidin （H） have an antioxidant property and quench free radicals. This study aimed to clarify the protective role of β-carotene and hesperidin as natural antioxidants on IM induced toxicity in hematological parameters and markers of cardiac muscle activity in male albino rats. The treatment of rats with IM showed a significant decrease in hemoglobin （Hb %）, MCH （mean corpuscular hemoglobin）, MCHC （mean corpuscular hemoglobin concentration）, HCT （hematocrit） values and RBCs count comparing with control group. On the other hand, MCV （mean corpuscular volume）, WBCs （white blood cells） and Pits （platelets） count pronounced a significant increase in IM group comparing to control. Also, αFP （plasma alpha fetoprotein）, CEA （carcinoembryonic antigen）, CK （creatine kinase）, CK-MB （creatine phosphokinase myocardial band） and LDH （lactate dehydrogenase） clarify a significant increase in IM group comparing to control. Both β-carotene and hesperidin mitigate the deleterious effects of IM on previous parameters. β-Carotene and hesperidin may protect hematopoietic system and heart muscle against toxicity of IM. These improvements of the results clarify the protective effect of the used antioxidants. Conclusion： β-carotene and hesperidin, natural antioxidants, have a protective effect against IM evoked hematological and biochemical changes.

Nutrient and Nutraceutical Potentials of Seaweed Biomass Ulva lactuca and Kappaphycus alvarezii

The study aimed at evaluating the nutrient and nutraceutical potentials of the selected seaweed biomass namely Ulva lactuca and Kappaphycus alvarezii. The selected seaweeds were found to be rich in macro and micronutrients. Kappaphycus contained high amount of ash when compared to U. lactuca. It is revealed that the mercury level in U. lactuca was 0.017 ppm and it was not detected in K. alvarezii. The heavy metal concentration in the selected seaweeds was within the tolerable value reported as quality criteria for edible seaweeds. These seaweeds were also found to be rich sources of vitamin antioxidants namely Vitamin C, total carotenoid, beta carotene, total chlorophyll and polyphenol. The phytochemical screening of the selected seaweeds showed that both seaweeds contained carbohydrates, protein, gums and mulicage, phenols, starch and quinones. Alkaloids was absent in Kappaphycus, saponins was moderate in U. actuca when compared to Kappaphycus. Glycosides, flavonoids, tannins and terpenoids were absent in both the species. The selected seaweeds also possess antibacterial activity. The selected biomass could be recommended for use in dietaries to combat protein energy malnutrition and in particular micronutrient deficiencies.

Stability of β-carotene microcapsules with Maillard reaction products derived from whey protein isolate and galactose as coating materials

The stability of β-carotene microcapsules using Maillard reaction products（MRPs） derived from whey protein isolate（WPI） and galactose as coating materials, was studied under the varying environmental conditions of temperature, pH, air, incandescent light, and ultraviolet（UV） light.Scanning electron microscopy showed that microcapsules prepared by WPI-galactose MRPs displayed a smooth and less concave-convex surface and that the particle size（D_（50）） of the microcapsules made with WPI-galactose MRPs was smaller than those made with WPI-galactose mixture.The storage stability of β-carotene microencapsulated in WPI-galactose MRPs was remarkably better than that of β-carotene microencapsulated in the WPI-galactose mixture and that of β-carotene crystal, in respect of temperature, pH, air, incandescent light, and UV light measurements.When the storage temperature was increased from 5 to 105 ℃, the retention rate of β-carotene microcapsules significantly decreased（P〈0.05）.When p H values were increased from 1 to 12, the β-carotene retention rate of the microcapsules significantly increased and afterward decreased.Compared with the retention rate of β-carotene microencapsulated in a WPI-galactose mixture, the retention rate of β-carotene microencapsulated in WPI-galactose MRPs was at a maximum between pH 8 and 9.Under the actions of air, incandescent light, and UV light, the retention rates of β-carotene microcapsules in WPI-galactose MRPs and WPI-galactose mixture, as well as in β-carotene crystal, decreased significantly as the storage time increased（P〈0.05）.Therefore, the use of WPI-galactose MRPs as coating materials can aid in improving the storage stability of β-carotene microcapsules.

Microencapsulated β-carotene preparation using different drying treatments

β-Carotene is one of the most abundant natural pigments in foods;however,usage of β-carotene is limited because of its instability.Microencapsulation techniques are usually applied to protect microencapsulated p-carotene from oxidization.In this study,β-carotene was microencapsulated using different drying processes:spray-drying,spray freeze-dryi ng,coati ng,and spray granulation.The properties of morphology,particle size,water con tent,thermal characteristic,and chemical stability have been explored and compared.Scanning electron microscopy measure?ments showed that the coated powder had a dense surface surro un ded by starch and suggested that the coati ng process gave a microencapsulated powder with the smallest bulk density and the best compressibility among the prepared powders.The chemical stabilities of microcapsules were evaluated during six months of storage at different temperatures.The coated powder had the highest mass fraction of β-carotene,which indicated that the coating process was superior to the three other drying processes.

Function of terahertz spectra in monitoring the decomposing process of biological macromolecules and in investigating the causes of photoinhibition

Chlorophyll α and β-carotene play an important role in harvesting light energy, which is used to drive photosynthesis in plants. In this study, terahertz(THz) and visible range spectra of chlorophyll α and β-carotene and their changes under light treatment were investigated. The results show that the all THz transmission and absorption spectra of chlorophyll α and β-carotene changed upon light treatment, with the maximum changes at 15 min of illumination indicating the greatest changes of the collective vibrational mode of chlorophyll α and β-carotene. The absorption spectra of chlorophyll α in the visible light region decreased upon light treatment, signifying the degradation of chlorophyll a molecules. It can be inferred from these results that the THz spectra are very sensitive in monitoring the changes of the collective vibrational mode, despite the absence of changes in molecular configuration. The THz spectra can therefore be used to monitor the decomposing process of biological macromolecules; however, visible absorption spectra can only be used to monitor the breakdown extent of biological macromolecules.