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1.
Carotenoids from carrots and tomatoes were separated with high-performance liquid chromatography (HPLC) and capillary supercritical fluid chromatography (SFC). All trans alpha- and beta-carotene were separated from their respective cis-isomers with capillary SFC. Carotenoids extracted from tomatoes included xanthophyll, lycopene and beta-carotene, while alpha- and beta-carotene were extracted from carrots. The HPLC separations were accomplished isocratically with a 25-cm column containing 5-microns ODS and methanol-acetonitrile-chloroform (47:47:6) or acetonitrile-dichloromethane (80:20). beta-Carotene cis-isomers were separated with SFC with a SB-cyanopropyl-25-polymethylsiloxane column, while alpha-carotene isomers were separated with two SB-cyanopropyl-50-polymethylsiloxane columns. Carotenoids from carrots and tomatoes were separated with a SB-phenyl-50-polymethylsiloxane column. Carbon dioxide with 1% ethanol was the SFC mobile phase. The eluent was monitored at 461 nm for HPLC and either 453 or 461 nm for SFC.  相似文献   

2.
高效液相色谱法测定藻类中的类胡萝卜素和叶绿素   总被引:16,自引:0,他引:16  
袁建平  张义明  史贤明  龚贤弟  陈峰 《色谱》1997,15(2):133-135
提出了用高效液相色谱法测定藻类中类胡萝卜素和叶绿素的方法。采用丙国等有机溶剂提取藻类中的类胡萝卜素和叶绿素,然后在反相C18柱上进行分离。流动相选用二氯甲烷/乙腈/甲醇/水(22.5:9.5:67.5:0.5),流速为1.0mL/min。用光度检测器检测报长为450um。叶黄素、α-胡萝卜素、β-胡萝卜素、叶绿素a和叶绿素b的平均回收串分别为99.1%,98.5%,99.4%,100.6%和99.9%,相对标准偏差分别为2.4%,5.6%,6.0%,4.1%和4.0%。  相似文献   

3.
High-performance liquid chromatography with photodiode array detection was developed for the separation and identification of carotenoids from a new sweet orange, Earlygold. Carotenoid pigments were extracted using hexane-acetone-ethanol and saponified using 10% methanolic potassium hydroxide. More than 25 carotenoid pigments were separated within 40 min using a ternary gradient (acetonitrile-methanol, methyl tert-butyl ether and water) elution on a C30 reversed-phase column. The carotenoid pattern of Earlygold was generally similar to the early season Hamlin but with some quantitative differences, especially with violaxanthin. Major carotenoids including violaxanthin, lutein, beta-cryptoxanthin, antheraxanthin, luteoxanthin, zeaxanthin, beta-carotene, and alpha-carotene were identified based on on-line spectral data obtained by a photodiode array detector, and comparison to the spectra of the standards and reported values. A numerical notation, the ratio of the peak heights between absorption bands, was also calculated to compare to the literature values.  相似文献   

4.
反相高效液相色谱法同时测定番茄中5种类胡萝卜素   总被引:17,自引:0,他引:17  
王强  韩雅珊  戴蕴青 《色谱》1997,15(6):534-536
用反相高效液相色谱法同时测定了番茄中的α-胡萝卜素、β-胡萝卜素、γ-胡萝卜素、番茄红素及斑蝥黄素。采用11(V/V)丙酮-石油醚混合液直接提取,提取液减压蒸干后,用甲醇溶解,经μ-BondapakC18色谱柱,以乙腈-二氯甲烷-甲醇(85105)为流动相,在450nm下进行检测,其保留时间依次为15.25,16.12,12.87,10.38和5.37min,最小检出限分别为0.25,0.22,0.18,0.14和0.05mg/L。  相似文献   

5.
Vitamin A deficiency is a health problem in Southeast Asia that can be corrected by feeding orange fruits and vegetables such as mango. Pumpkin is a traditional Korean food that is easy to store and is already believed to have health benefits. We extracted carotenoids from pumpkin by liquid-liquid extraction and by supercritical fluid extraction. We measured carotenoids by reversed-phase chromatography with diode array detection. The major carotenoid in pumpkin (> 80%) is beta-carotene, with lesser amounts of lutein, lycopene, alpha-carotene and cis-beta-carotene. Pumpkin is a rich source of beta-carotene and might be useful for preventing Vitamin A deficiency.  相似文献   

6.
A single laboratory validation (SLV) study was conducted for a liquid chromatography (LC) method for the determination of total and all-trans-beta-carotene in a variety of dietary supplements, including multivitamin tablets, softgels, capsules, and beadlet raw materials. Extraction variants were developed for the different types of supplements tested based upon the supplement type and level of beta-carotene. Water dispersible formulations such as powders, emulsions, tablets, and capsules were enzymatically digested with protease and extracted with dichloromethane-ethanol. Oily suspensions were directly dissolved in dichloromethane-ethanol. After appropriate dilution or concentration, the extracts were chromatographed by using either a reversed-phase C18 column or, in products containing high amounts of alpha-carotene, a reversed-phase C30 column. The LC systems provided linear responses in the range of 0.1-50 microg beta-carotene/mL. The main geometrical isomers of beta-carotene (all-trans, 9-cis, 13-cis, and 15-cis) were well separated from each other and from other carotenoids such as a-carotene, cryptoxanthin, lutein, lycopene, and zeaxanthin. Duplicate determinations of total beta-carotene performed by 2 technicians in 8 different test materials on 5 different days resulted in relative standard deviations of 1.2-4.4%. Recoveries determined for supplements and beadlet raw material spiked with beta-carotene levels of 10 microg to 100 mg/test portion and 0.2-40%, respectively, ranged from 97.5 to 102.1%. On the basis of the accuracy, precision, and recovery results from the SLV study, the method is suggested for a collaborative study on the determination of total and all-trans-beta-carotene in dietary supplements.  相似文献   

7.
We used reversed-phase HPLC with diode array detection to simultaneously measure the major isoforms of vitamins A, E, and the carotenoids in serum from 55 healthy people with spinal cord injuries. Typically, the method measured retinol (vitamin A), alpha-tocopherol (vitamin E) and beta-carotene, alpha-carotene, lutein, lycopene, and cryptoxanthin (carotenoids). gamma-Tocopherol (vitamin E), 25-hydroxycalciferol (vitamin D), and the carotenoid zeaxanthin could also be measured when they were present in high concentrations. Healthy people with spinal cord injuries were more likely than similar people without injuries to have low concentrations of alpha-tocopherol, and to a lesser extent retinol and beta-carotene.  相似文献   

8.
Theoretical study of the enthalpies of formation (DeltaHf) for polyenes up to nine ethylene units and for several C40H56 carotenes including beta-carotene, alpha-carotene, lycopene, and prolycopene is presented. For polyenes and small branched alkenes, we used G2, G3, and G3MP2B3 theories, and the DeltaHf values were evaluated with the atomization, isodesmic bond separation, and homodesmic schemes. The applicability of six DFT functionals were evaluated by comparing their predictions with those obtained using G3 theory within the atomization scheme. Additivity approaches, including atom equivalents and group equivalents using DFT and semiempirical theories, were explored. We found that group equivalents associated with isodesmic reactions are able to provide the most accurate predictions within the test set. The predictions from the six functionals are in good agreement with the G3 results. Among them, B3LYP performs the best, with an average absolute deviation of only 0.30 kcal/mol. The application of DFT in the prediction for the DeltaHf value of C40H56 carotenes is promising.  相似文献   

9.
Carotenoids and vitamins A and E in multivitamin tablets can be determined simultaneously by reversed-phased liquid chromatography (LC) with a programmable UV detector. Samples were dissolved in dimethyl sulfoxide and then extracted with hexane. A portion was injected onto a Symmetry C18, 150 x 4.6 mm id, 5 microns column and chromatographed with a mobile phase of acetonitrile--0.25% ammonium acetate in methanol and 0.05% triethylamine in dichloromethane. A step gradient was used. The system was operated at 25 degrees C with a flow rate of 1.5 mL/min. UV detection was at 325 nm for retinols, 285 nm for tocopherols, and 450 nm for carotenoids. Detection limits were less than 0.3 ng for retinol and retinyl acetate; 2 ng for alpha-tocopherol acid succinate; 10 ng for alpha-tocopherol, gamma-tocopherol, and alpha-tocopherol acetate; and 0.4 ng for alpha-carotene and beta-carotene. Intraday and interday coefficients of variation ranged from 1.40 to 5.20%. The sample preparation method and LC assay are practical for quality control and routine analysis of multivitamin tablets.  相似文献   

10.
Calculation of the HPLC chromatographic retention times of different carotenoids supported our improved chromatographic separation of beta-carotene and lutein in four tomatoes and two tomatillo varieties in fresh form and after three different cooking procedures: pot boiling, cooking in a pressure cooker and microwaving. A good separation was achieved experimentally using an Ultrasphere ODS column and gradient elution with an acetonitrile-tetrahydrofuran-water mobile phase. It was shown that diverse tomato species contained different amounts of beta-carotene (6-400 microg/100 mg) and lutein (2-30 microg/100 mg). The concentration in fresh samples was higher than in cooked tomatoes. The beta-carotene content in fresh tomatillo varied between 2 and 20 microg/100 mg. Microwaving caused partial destruction of the beta-carotene and lutein in tomatillos.  相似文献   

11.
Among more than 50 provitamin carotenoids, beta-carotene is the most metabolically active source of retinol. Despite diets rich in fruits and vegetables containing beta-carotene, vitamin A deficiency is the leading cause of blindness and childhood mortality in developing countries. In addition, the uncertainty of beta-carotene bioconversion into vitamin A suggests that new data are needed to update the nutritional guidelines in developed countries. Previously, we reported the development of a carotene/retinol plateau isotopic enrichment method (CarRet PIE) for the determination of beta-carotene bioavailability and bioconversion into retinol, which utilizes positive ion atmospheric pressure chemical ionization (APCI) liquid chromatography/mass spectrometry (LC/MS). While seeking to validate the CarRet PIE using a mass balance approach requiring fecal measurements of beta-carotene and retinol, interference was encountered that required substantial modifications of the LC/MS assay. Here we report a new LC/MS assay that is based on the detection of molecular anions of beta-carotene using negative ion APCI with a reversed-phase C30 column for HPLC separation. Sample preparation required saponification to eliminate interfering triglycerides. The limit of detection (LOD) of beta-carotene was 0.25 pmol calculated on the basis of an injection of 20 microL of 0.0125 microM beta-carotene, and the limit of quantitation (LOQ) was 1.0 pmol based on the injection of 20 microL of 0.050 microM beta-carotene. The linear range was 1.1 to 2179 pmol on-column. The wide linear range and low LOD and LOQ of this assay facilitated the sensitive and selective quantitative analysis of beta-carotene in both serum and fecal samples in support of an on-going clinical investigation of beta-carotene bioavailability and bioconversion into vitamin A.  相似文献   

12.
Highly transparent silica-surfactant nanocomposite films containing photosynthetic pigments have been successfully formed through the solubilization of chlorophyll a (Chl a) into surfactant micelles. The UV-vis absorption spectra indicated that a large amount of Chl a were transformed into pheophytin a in the films. These photosynthetic pigments were well dispersed in the surfactant assemblies and their chlorin rings were exposed to the surface of silica layers. Even under an air atmosphere, the photostability of immobilized pigments was largely improved in comparison with that in a homogeneous Chl a solution. Because both Chl a and pheophytin a molecules are effective for the photosensitive charge separation, the present film system is very suitable for heterogeneous immobilizing media for photosynthetic pigments from the viewpoint of in vitro biomimetic devices for solar energy conversion.  相似文献   

13.
In 1997, the National Institute of Standards and Technology (NIST) released Standard Reference Material (SRM) 2383 Baby Food Composite. This SRM can be used as a control material when assigning values to in-house control materials and when validating analytical methods for the measurement of proximates, vitamins, minerals, and trace elements in baby foods and similar matrixes. The Certificate of Analysis for SRM 2383 provides certified and reference values for concentrations of lutein, zeaxanthin, beta-cryptoxanthin, lycopene, alpha-carotene, beta-carotene, delta-tocopherol, gamma-tocopherol, alpha-tocopherol, retinol, and retinyl palmitate for 2 types of sample preparation--extraction and saponification. The assigned values were based on the agreement of measurements made by NIST and collaborating laboratories. The Certificate of Analysis also provides reference and information values for concentrations of proximates, minerals, and additional vitamins; assignment of these values is discussed in a companion paper (this issue, page 276).  相似文献   

14.
In this paper the application of on-line HPLC-UV-APCI (atmospheric pressure chemical ionization) mass spectrometry (MS) coupling for the separation and determination of different carotenoids as well as cis/trans isomers of beta-carotene is reported. All HPLC separations were carried out under RP conditions on self-synthesized polymeric C30 phases. The analysis of a carotenoid mixture containing astaxanthin, canthaxanthin, zeaxanthin, echinenone and beta-carotene by HPLC-APCI-MS was achieved by scanning the mass range from m/z 200 to 700. For the characterization of a sample containing cis/trans isomers of beta-carotene as well as their oxidation products, a photodiode-array UV-visible absorbance detector was used in addition between the column and the mass spectrometer for structural elucidation of the geometrical isomers. The detection limit for beta-carotene in positive-ion APCI-MS was determined to be 1 pmol. In addition, an extract of non-polar substances in vegetable juice has been analyzed by HPLC-APCI-MS. The included carotenoids could be identified by their masses and their retention times.  相似文献   

15.
An improved separation method for chlorophyll metabolites in Oriental tobacco leaf was developed. While Oriental leaf still gives the green color even after the curing process, little attention has been paid to the detailed composition of the remaining green pigments. This study aimed to identify the green pigments using non aqueous reversed phase chromatography (NARPC). To this end, liquid chromatograph (LC) equipped with a photo diode array detector (DAD) and an atmospheric pressure chemical ionization/mass spectrometer (APCI/MSD) was selected, because it is useful for detecting low polar non-volatile compounds giving green color such as pheophytin a. Identification was based on the wavelength spectrum, mass spectrum and retention time, comparing the analytes in Oriental leaf with the commercially available and synthesized components. Consequently, several chlorophyll metabolites such as hydroxypheophytin a, solanesyl pheophorbide a and solanesyl hydroxypheophorbide a were newly identified, in addition to typical green pigments such as chlorophyll a and pheophytin a. Chlorophyll metabolites bound to solanesol were considered the tobacco specific components. NARPC expanded the number of detectable low polar chlorophyll metabolites in Oriental tobacco leaf.  相似文献   

16.
Self-assembling of a carotenoid and pheophytin a into a supramolecular system was observed on the surface of nanocrystalline TiO2, and the photoinduced electron-transfer reactions within the system were studied by means of femtosecond transient absorption and laser flash photolysis techniques. Excitation of the pheophytin moiety results in ultrafast electron transfer from carotenoid to the excited pheophytin, creating a long-lived charge-separated state. Two decay pathways of the formed pheophytin a anion radical are proposed. The first is a direct back electron recombination forming a carotenoid triplet state on the nanosecond time scale, while the other is suggested to occur via electron injection to the TiO2 nanoparticle. These results demonstrate that a self-assembled carotenoid-pheophytin system leads to an efficient reductive quenching of the pheophytin moiety, suggesting that a similar mechanism can operate also in natural photosynthetic systems. Moreover, the formation of a long-lived charge-separated state indicates that such self-assembling strategy may be also considered for novel dye-sensitized solar cell constructions and other artificial systems aiming to mimic the electron-transfer chain in natural photosynthesis.  相似文献   

17.
An isocratic high-performance liquid chromatographic method specifically developed to allow simple and rapid determination of beta-carotene concentrations in serum and plasma is reported. Using a method modified from a previously published technique, serum and plasma proteins are denatured by exposure to perchloric acid, and beta-carotene is subsequently extracted into an organic matrix consisting of ethyl acetate-tetrahydrofuran (1:1); no evaporation step is required. Separation is achieved using isocratic elution from a reversed-phase C18 column with UV detection at 436 nm. Recovery of beta-carotene from water and plasma was greater than 98.1%; beta-carotene was stable in the extraction matrix for at least 4 h. Three anticoagulants (oxalate, citrate, and EDTA) caused losses of beta-carotene; perchloric acid and tetrahydrofuran could also destroy beta-carotene under certain conditions. Each run required less than 15 min; within-day coefficient of variation for identical samples averaged 2.3%, between-day coefficient of variation was 4.4% and sensitivity was better than 10 ng/ml. Stability of beta-carotene in plasma was also examined. This method permits a simple, rapid, sensitive, precise, and accurate determination of beta-carotene using 0.5 ml of serum or heparinized plasma.  相似文献   

18.
Twelve laboratories representing 4 countries participated in an interlaboratory study conducted to determine all-trans-veta-carotene and total beta-carotene in dietary supplements and raw materials. Thirteen samples were sent as blind duplicates to the collaborators. Results obtained from 11 laboratories are reported. For products composed as softgels and tablets that were analyzed for total beta-carotene, the reproducibility relative standard deviation (RSDR) ranged from 3.35 to 23.09% and the HorRat values ranged from 1.06 to 3.72. For these products analyzed for trans beta-carotene, the reproducibility relative standard deviation (RSDR) ranged from 4.28 to 22.76% and the HorRat values ranged from 0.92 to 3.37. The RSDr and HorRat values in the analysis of a beadlet raw material were substantial and it is believed that the variability within the material itself introduced significant variation in subsampling. The method uses high pressure liquid chromatography (LC) in the reversed-phase mode with visible light absorbance for detection and quantitation. If high levels of alpha-carotenes are present, a second LC system is used for additional separation and quantitation of the carotene species. It is recommended that the method be adopted as an AOAC Official Method.  相似文献   

19.
The extraction of carotenoids from Japanese persimmon peels by supercritical fluid extraction (SFE), of which the solvent was CO(2), was performed. In order to enhance the yield and selectivity of the extraction, some portion of ethanol (5 - 20 mol%) was added as an entrainer. The extraction temperature ranged from 313 to 353 K and the pressure was 30 MPa. The effect of temperature on the extraction yield of carotenoids was investigated at 10 mol% of the ethanol concentration in the extraction solvent, and a suitable temperature was found to be 333 K among the temperatures studied with respect to the carotenoid yield. With increasing the entrainer amount from 0 to 10 mol% at a constant temperature (333 K), the carotenoid yield in the extraction was improved, whereas the selectivity of the extracted carotenoids was drastically depressed. We also conducted qualitative and quantitative analyses for the carotenoid components in the extract by HPLC, and analyzed the extraction behavior of each individual carotenoid (alpha-carotene, beta-carotene, beta-cryptoxanthin, lycopene, lutein, and zeaxanthin). The selectivity of each carotenoid changed with the elapsed time and its time evolution was dependent on the carotenoid component, indicating that the location profile and the content can be important factors to understand the SFE behavior of each carotenoid in persimmon peels.  相似文献   

20.
Abstract— In the first part of this study the spectral properties of pheophytin a in rigid, unstretched anhydrous polyvinyl alcohol and nitrocellulose films have been studied in order to establish the influence of the central magnesium atom on the state of chlorophylls in polymer systems. The absorption, fluorescence, excitation spectra and fluorescence intensity decays in the polymer films and in the solutions from which they are cast are reported. It is shown that pheophytin a aggregate formation is influenced by the nature of the polymer system. An aggregate of pheophytin a is found in polyvinyl alcohol films over a wide concentration range. On the other hand, pheophytin a exists in the monomeric form in unstretched nitrocellulose films at concentrations below 6 × 10-6 mol/g.
In the second part of this work, the influence of stretching of the films on the state and distribution of embedded chlorophyll pigments, is described. Here we show that the chlorophyll a molecules are found to undertake a heterogenous distribution in polyvinylalcohol matrices, since stretching partially disrupts the pocket-like structures present in unstretched films. In contrast, chlorophyll a and pheophytin a molecules can be embedded in a monomeric state in nitrocellulose matrices and moreover they remain homogeneously distributed upon stretching. The chlorophyll/nitrocellulose system is concluded to be a useful model system for studies of donor-donor energy transfer processes.  相似文献   

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