Application of near-infrared reflectance spectroscopy to the evaluation of rutin and D-chiro-Inositol contents in tartary buckwheat

Tartary buckwheat [Fagopyrum tataricum (L.) Gaench] is rich in rutin and D- chiro-inositol (DCI), which have beneficial effects in the treatment of hemorrhagic diseases and insulin-resistant diseases, respectively. The current methods of extraction and detection of rutin and DCI are complex and time-consuming; a simple way of analyzing these compounds in the native matrix would be desirable. In this work, near-infrared reflectance spectroscopy (NIRS) was applied to determine the contents of rutin and DCI in tartary buckwheat. The spectral data were compared with those determined from high-performance liquid chromatography (HPLC) methods. Models for predicting rutin and DCI contents in buckwheat were developed using a partial least-squares algorithm. Cross-validation procedures indicated good correlations between HPLC data and NIRS predictions (R2 = 0.76 for rutin and R2 = 0.86 for DCI). The rutin content ranged from 0.998 to 1.75%, while the DCI content covered 0.179-0.200%. The results showed that NIRS, a well-established and widely applied technique, could be applied to determine rutin and DCI in tartary buckwheat rapidly and nondestructively.     

Front-surface absorbance spectra of wheat flour: determination of carotenoids

Front-surface absorbance spectra of wheat flours in the 250-650 nm region can be obtained by measuring reflectance spectra with a conventional spectrofluorometer suitably set to detect light scattered from powder samples. The spectra recorded on flour samples, obtained from seeds of four bread and five durum wheats, show high-intensity absorption bands due to aromatic amino acids of wheat proteins and low-intensity bands due to chromophores bound to low-molecular-weight compounds. The intensity of these last bands is proportional to the concentration of the corresponding chromophores present in the flour; thus, it can be used to measure the content of the compounds containing the chromophore(s). In particular, a quantitative determination of the carotenoids actually present in the flours is made, obtaining information on the original content of the seeds. This determination is important, as, for example, xanthophylls are well-known antioxidants and free-radical scavengers involved in aging processes of seeds. Reflectance measurements on powder samples are far more economic in terms of time and materials consumption than methods such as extraction and HPLC analysis of extracts and, in addition, give an evaluation of the overall content of carotenoids with absorption bands in the spectral range 450-500 nm. Application of the technique to other food powders with low-intensity absorption bands in the near-UV and vis region is possible.     

Evaluation of analytical methods for carotenoid extraction from biofortified maize (Zea mays sp.)

Biofortification of maize with beta-carotene has the potential to improve vitamin A status in vitamin A deficient populations where maize is a staple crop. Accurate assessment of provitamin A carotenoids in maize must be performed to direct breeding efforts. The objective was to evaluate carotenoid extraction methods and determine essential steps for use in countries growing biofortified maize. The most reproducible method based on coefficient of variation and extraction efficiency was a modification of Kurilich and Juvik (1999). Heat and saponification are required to release carotenoids from biofortified maize and remove oils interfering with chromatographic analysis. For maize samples with high oil content, additional base may be added to ensure complete saponification without compromising results. Degradation of internal standard before carotenoids were released from the maize matrix required the addition of internal standard after heating to prevent overestimation of carotenoids. This modified method works well for lutein, zeaxanthin, beta-cryptoxanthin, alpha-carotene, and beta-carotene.     

Diversity in Phytochemical Composition and Antioxidant Capacity of Dent,Flint, and Specialty Corns

Dent, flint, and specialty genotypes of Indian yellow maize were evaluated for phytochemical content and their hydrophilic antioxidant capacity. Solid‐phase extraction coupled with solid–liquid extraction was used to obtain free phenolics and flavonoids from maize samples, reducing the use of excessive solvents and handling time. Bound phenolics were extracted with enhanced acidic hydrolysis to improve extractability. The phenolic contents in Indian maize genotypes ranged from 46 to 79 μmol of ferulic acid equivalents per gram of sample on a dry basis (db). Carotenoids in Indian maize genotypes were found to be equivalent to Chinese yellow maize and ranged from 13 to 24 μg/g of sample (db), whereas tocol content varied greatly in the range of 607–1,238 μg of α‐tocopherol equivalents/g of sample (db). Dent and flint corn did not exhibit differences (P > 0.05) in their phenolic contents, whereas among the specialty genotypes sweet corn contained the highest phenolics but least carotenoids (P < 0.05). Bound extracts appeared to contribute largely to 2,2′‐diphenyl‐1‐picrylhydrazyl free radical scavenging, hydrogen peroxide scavenging, and ferric reducing antioxidant power of maize genotypes. Sweet corn exhibited higher antioxidant capacity (P < 0.05) among all the genotypes. The antioxidant capacities correlated well with the total phenolic contents of the maize genotypes.     

Near-infrared reflectance spectroscopy (NIRS) for protein, tryptophan, and lysine evaluation in quality protein maize (QPM) breeding programs

Quality protein maize (QPM) has approximately twice the tryptophan (Trp) and lysine (Lys) concentrations in protein compared to normal maize. Because several genetic systems control the protein quality of QPM, it is essential to regularly monitor Trp and/or Lys in breeding programs. Our objective was to examine the potential of near-infrared reflectance spectroscopy (NIRS) to enhance the efficiency of QPM research efforts by partially replacing more expensive and time-consuming wet chemistry analysis. More than 276 maize samples were used to develop NIRS models for protein content (PC), Trp, and Lys. The standard error of prediction (SEP) for the calibration and the coefficient of determination for validation (R(2)(v)) were 0.26 and 0.96 for PC, 0.005 and 0.85 for Trp, and 0.02 and 0.75 for Lys. When the NIRS models were used to evaluate 266 S2 lines from five QPM breeding populations, the coefficients of determination between NIRS and the chemical data were 0.94, 0.76, and 0.80 for PC, Trp, and Lys, respectively. Therefore, the NIRS models can be used to support the QPM breeding efforts.     

Nondestructive prediction of total phenolics, flavonoid contents, and antioxidant capacity of rice grain using near-infrared spectroscopy

Phytochemicals such as phenolics and flavonoids, which are present in rice grains, are associated with reduced risk of developing chronic diseases such as cardiovascular disease, type 2 diabetes, and some cancers. The phenolic and flavonoid compounds in rice grain also contribute to the antioxidant activity. Biofortification of rice grain by conventional breeding is a way to improve nutritional quality so as to combat nutritional deficiency. Since wet chemistry measurement of phenolic and flavonoid contents and antioxidant activity are time-consuming and expensive, a rapid and nondestructive predictive method based on near-infrared spectroscopy (NIRS) would be valuable to measure these nutritional quality parameters. In the present study, calibration models for measurement of phenolic and flavonoid contents and antioxidant capacity were developed using principal component analysis (PCA), partial least-squares regression (PLS), and modified partial least-squares regression (mPLS) methods with the spectra of the dehulled grain (brown rice). The results showed that NIRS could effectively predict the total phenolic contents and antioxidant capacity by PLS and mPLS methods. The standard errors of prediction (SEP) were 47.1 and 45.9 mg gallic acid equivalent (GAE) for phenolic content, and the coefficients of determination ( r (2)) were 0.849 and 0.864 by PLS and mPLS methods, respectively. Both PLS and mPLS methods gave similarly accurate performance for prediction of antioxidant capacity with SEP of 0.28 mM Trolox equivalent antioxidant capacity (TEAC) and r (2) of 0.82. However, the NIRS models were not successful for flavonoid content with the three methods ( r (2) < 0.4). The models reported here are usable for routine screening of a large number of samples in early generation screening in breeding programs.     

Different effects of microwave and ultrasound on the stability of (all-E)-astaxanthin

Both microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE) have been widely applied in the extraction of a variety of biologically active compounds including carotenoids due to their lower pollution to environment, high extraction efficiency, lower cost, and shorter extraction time as compared to conventional extraction techniques. However, there are few reports on their effects on the stability of these compounds. In the present study, the stability of (all-E)-astaxanthin, one of the carotenoids, was examined under the action of both ultrasound and microwave. Results showed that microwave induced the isomerization of (all-E)-astaxanthin to its Z analogues, preferentially to (13Z)-astaxanthin as analyzed by HPLC coupled with diode array detection and LC-MS; and the percentage of the isomerization increased with increasing both treatment time and microwave power. In contrast to the microwave, the ultrasound degraded (all-E)-astaxanthin to unidentified colorless compound(s) as suggested by HPLC analysis and UV/vis measurements, and the degradation likewise increased as both treatment time and ultrasonic power increased. The results presented here emphasized that both MAE and UAE techniques should be carefully used in the extraction of unstable compounds such as (all-E)-astaxanthin.     

Multivariate analysis of antioxidant power and polyphenolic composition in red wines

It has been demonstrated that the dietary intake of compounds having antioxidant activity is very important, and various chemical, biological, and electrochemical methods have been proposed to evaluate the antioxidant power of compounds such as polyphenols. Wine, although nonessential, has a high polyphenol content up to 2-3 g/L in red wines obtained by traditional maceration. The polyphenol content of wines is usually evaluated by the Folin reagent, which provides an appropriate response to the requirements of wine manufacturers. Because the presence of individual polyphenols may be evaluated by HPLC, more or less selective methods toward the various chemical classes of polyphenols have been developed. An HPLC method set up recently was applied to evaluate how individual polyphenols contributed to the overall antioxidant power (AOP) of 60 Italian red wines, trying to identify the effect that individual compounds may have on the total AOP. Application of the multivariate analysis allowed us to detect some determining compounds such as gallic acid and some flavonols. On the basis of the correlation between two traditional chemical methods, namely the total polyphenol determination by the Folin reagent and the flavanol determination by the condensation reaction with p-(dimethylamino)-cinnamaldehyde, it was shown that the use of these two merely chemical methods is well correlated (r = 0.83 and 0.87) to an AOP evaluation of red wines.     

Development of a new method for the complete extraction of carotenoids from cereals with special reference to durum wheat (Triticum durum Desf.)

Due to the growing interest in the role of carotenoids in human health, their qualitative and quantitative analysis in foods is becoming more and more important. High-performance liquid chromatography has become the method of choice for the determination of these phytochemicals. A crucial step prior to the chromatographic separation is the quantitative extraction from the food matrix which was proven to be impeded in durum wheat. To optimize the extraction procedure, several factors with influence on extractability of carotenoids were investigated. Finally, it was shown that soaking of samples in water for 5 min prior to extraction with organic solvents had the strongest impact on extraction yield and led to the most rapid and gentle method. Contents of carotenoids in the extracts of several durum wheat and corn samples were doubled by soaking in water before extracting with methanol/tetrahydrofuran (1/1, v/v). In light of these findings, literature data on contents of carotenoids in cereal grains have to be viewed critically regarding the extraction procedures employed.     

油用牡丹单粒种子含油量NIRS模型的建立

为了建立油用牡丹单粒种子含油量的近红外测定模型,便于高含油量单株的选育,采用索氏抽提法测试了200份油用牡丹凤丹单粒种子的含油量,并应用近红外反射光谱技术(NIRS)采集了200份样品的光谱数据,通过偏最小二乘法(PLS)和主成分回归法(PCR)构建了油用牡丹单粒种子含油量的数学模型。结果表明,索氏抽提法中,均匀粉碎后的油用牡丹籽样品干燥烘焙条件为105℃ 2 h,牡丹籽抽提时间为20 h,测出的含油量变化范围在10%~28%之间,籽油含量基本符合正态分布。NIRS法构建的模型最佳参数为:采用PLS法,光程固定,一阶导数消除背景,数据平滑处理采用Norris derivative filter的方法,平滑参数选用5和3。内部交叉检验校正相关系数r₁为0.980 1、预测相关系数r₂为0.957 6、校正均方根误差(RMSEC)为0.463、预测均方根误差(RMSEP)为0.705。外部检验相关系数达 0.957 6, 平均误差小于3%。本试验所构建的牡丹单粒种子含油量的NIRS模型可靠,可以用于分析油用牡丹单粒种子的含油量。     

Quantification of kavalactones and determination of kava (Piper methysticum) chemotypes using near-infrared reflectance spectroscopy for quality control in vanuatu

Kava ( Piper methysticum Forst f., Piperaceae) has anxiolytic properties and the ability to promote a state of relaxation without the loss of mental alertness. The rapid growth of the nutraceutical market between 1998 and 2000 has been stopped by a ban in Europe and Australia because of some suspicion of liver toxicity. It is now important to develop a fast, cheap, and reliable quality test to control kava exports. The aim of this study is to develop a calibration of the near-infrared reflectance spectroscopy (NIRS) using partial least-squares (PLS) regression. Two hundred thirty-six samples of kava roots, stumps, and basal stems were collected from the Vanuatu Agricultural Research and Technical Centre germplasm collection and from four villages. These samples, representing 45 different varieties, were analyzed using NIRS to record their absorption spectra between 400 and 2500 nm. A set of 101 selected samples was analyzed for their kavalactone content using HPLC. The results were used for PLS calibration of the NIRS. The NIRS prediction of the kavalactone content and the dry matter were in agreement with the HPLC results. There were good correlations between these two series of results, and coefficients ( R (2)) were all close to 1. The measurements were reproducible and had repeatability on par with the HPLC method. The NIRS system has been calibrated for the six major kavalactone content measurements, and it is suggested that this method could be used for quality control in Vanuatu.     

Screening and identification of antioxidants in biological samples using high-performance liquid chromatography-mass spectrometry and its application on Salacca edulis Reinw

In this study, a new approach was developed for screening and identifying antioxidants in biological samples. The approach was based on significant decreases of the intensities of ion peaks obtained from high-performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) upon reaction with 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals. HPLC-MS/MS was further applied to elucidate structures of antioxidant peaks characterized in a spiking test. The new approach could also be used to monitor the reactivity of antioxidants in biological sample with free radicals. The approach was successfully applied to the identification of antioxidants in salak (Salacca edulis Reinw), a tropical fruit that is reported to be a very good source of natural antioxidants, but it was still not clear which compounds were responsible for its antioxidant property. The antioxidants in salak were identified to be chlorogenic acid, (-)-epicatechin, and singly linked proanthocyanidins that mainly existed as dimers through hexamers of catechin or epicatechin. In salak, chlorogenic acid was identified to be an antioxidant of the slow reaction type as it reacted with free radicals much more slowly than either (-)-epicatechin or proanthocyanidins. The new approach was proved to be useful for the characterization and identification of antioxidants in biological samples as a mass detector combined with an HPLC separation system not only serves as an ideal tool to monitor free radical active components but also provides their possible chemical structures in a biological sample.     

On-line HPLC detection of tocopherols and other antioxidants through the formation of a phosphomolybdenum complex

An on-line method to detect and quantify antioxidant species in complex extracts has been developed as a combination of conventional HPLC separation and a postcolumn reaction with phosphomolybdenum reagent at acidic pH. Sample analytes were chromatographed by HPLC, and the postcolumn formation of a phosphate/Mo(V) complex was detected at 598 nm with an on-line absorbance detector. An optimized instrumental system was set up using pure alpha-tocopherol, and it was successfully tested with complex food extracts including lettuce, tomato, red pepper, and soybean seed, where several tocopherols and carotenoids were identified. A potential application of this detection method to quantitatively determine different antioxidants was considered, and a specific application to the determination of tocopherols was developed. The new method was characterized with respect to linearity interval, repeatability, and reproducibility, the quantitative results obtained were validated by comparison with a conventional HPLC method with fluorometric detection, and it was applied to the determination of tocopherols in different foods. The results suggest that the proposed on-line HPLC method can be a powerful instrument for the detection, purification, and characterization of natural antioxidants.     

Normal phase high-performance liquid chromatography method for the determination of tocopherols and tocotrienols in cereals

The eight vitamers of vitamin E (alpha-, beta-, gamma-, and delta-tocopherols and -tocotrienols) have different antioxidant and biological activities and have different distributions in foods. Some cereals, especially oat, rye, and barley, are good sources of tocotrienols. A fast procedure for the determination of tocopherols and tocotrienols (tocols) in cereal foods was developed. It involves sample saponification and extraction followed by normal phase high-performance liquid chromatography (HPLC). The results have been compared with those found by direct extraction without saponification. The method is sensitive and selective enough to be tested on a wide variety of cereal samples. The highest tocol levels were found in soft wheat and barley ( approximately 75 mg/kg of dry weight). beta-Tocotrienol is the main vitamer found in hulled and dehulled wheats (from 33 to 43 mg/kg of dry weight), gamma-tocopherol predominates in maize (45 mg/kg of dry weight) ), and alpha-tocotrienol predominates in oat and barley (56 and 40 mg/kg of dry weight, respectively).     

Near-Infrared Reflectance Spectroscopy (NIRS) assessment of δ(18)O and nitrogen and ash contents for improved yield potential and drought adaptation in maize

The oxygen isotope composition (δ(18)O), accumulation of minerals (ash content), and nitrogen (N) content in plant tissues have been recently proposed as useful integrative physiological criteria associated with yield potential and drought resistance in maize. This study tested the ability of near-infrared reflectance spectroscopy (NIRS) to predict δ(18)O and ash and N contents in leaves and mature kernels of maize. The δ(18)O and ash and N contents were determined in leaf and kernel samples from a set of 15 inbreds and 18 hybrids grown in Mexico under full irrigation and two levels of drought stress. Calibration models between NIRS spectra and the measured variables were developed using modified partial least-squares regressions. Global models (which included inbred lines and hybrids) accurately predicted ash and N contents, whereas prediction of δ(18)O showed lower results. Moreover, in hybrids, NIRS clearly reflected genotypic differences in leaf and kernel ash and N contents within each water treatment. It was concluded that NIRS can be used as a rapid, cost-effective, and accurate method for predicting ash and N contents and as a method for screening δ(18)O in maize with promising applications in crop management and maize breeding programs for improved water and nitrogen use efficiency and grain quality.     

近红外光谱法测定玉米秸秆饲用品质

为了对玉米秸秆的饲用品质进行可靠、便捷、快速的分析和评价,该研究以不同品种、密度、氮肥和水分处理的不同发育时期和不同部位玉米秸秆为试验材料,应用近红外光谱（NIRS）技术和偏最小二乘法（PLS）,采用一阶导数+中心化+多元散射校正的光谱数据预处理方法,构建了玉米秸秆体外干物质消化率（IVDMD）、酸性洗涤纤维（ADF）、中性洗涤纤维（NDF） 和可溶性糖（WSC）含量的NIRS分析模型。所建立的IVDMD、ADF、NDF和WSC含量的NIRS校正模型决定系数（R2cal）分别为0.9906、0.9870、0.9931和0.9802,交叉验证决定系数（R2cv）分别为0.9593、0.9413 、0.9678和0.9342,外部验证决定系数（R2val）分别为0.9549、0.9353、0.9519和0.9191,各项标准差（SEC、SECV和SEP）为0.935～1.904,相对分析误差（RPD）均大于3。结果表明,各参数的NIRS分析模型可用于玉米秸秆饲用品质的分析和品种选育的快速鉴定。     

Preparation of ferulic acid from agricultural wastes: its improved extraction and purification

Ferulic acid (FA) is a phenolic antioxidant present in plants, which is widely used in the food and cosmetic industry. In the present study, various agricultural wastes such as maize bran, rice bran, wheat bran, wheat straw, sugar cane baggasse, pineapple peels, orange peels, and pomegranate peels were screened for the presence of esterified FA (EFA). Among the sources screened, maize bran was found to contain the highest amount of EFA. Pineapple peels, orange peels, and pomegranate peels were also found to contain traces of EFA. Alkaline extraction of EFA from maize bran was carried out using 2 M NaOH. Response surface methodology (RSM) was used for optimization of EFA extraction, which resulted in a 1.3-fold increase as compared to the unoptimized conventional extraction technique. FA was analyzed by means of high-performance liquid chromatography (HPLC). Purification was carried out by adsorption chromatography using Amberlite XAD-16 followed by preparative high-performance thin-layer chromatography (HPTLC). The recovery of Amberlite XAD-16 purified FA was up to 57.97% with HPLC purity 50.89%. The fold purity achieved was 1.35. After preparative HPTLC, the maximum HPLC purity obtained was 95.35% along with an increase in fold purity up to 2.53.     

Determination of antioxidant activity of wine byproducts and its correlation with polyphenolic content

It has been demonstrated that wine and other products derived from the grape have a high antioxidant capability; as a possible consequence of this, they may have potential benefits for health. The byproducts of the winemaking process represent a source of antioxidant compounds that has been relatively unexploited to date, but that is now the subject of increasing industrial interest. This article describes an approach to the study of the antioxidant activity of grape marcs, stalks, and dregs of both white and red varieties. This activity is compared with the measurements of their content of total polyphenols and of individual polyphenolic compounds, identified and quantified by HPLC. From the results we have been able to establish a positive correlation between the antioxidant activity and the total polyphenolic content of samples, but not with specific compounds.     

Characterization of carotenoid pigments in mature and developing kernels of selected yellow-endosperm sorghum varieties

Sorghum is a critical source of food in the semiarid regions of sub-Saharan Africa and India and a potential source of dietary phytochemicals including carotenoids. The objective of this study was to determine the carotenoid profiles of sorghum cultivars, selected on the basis of their yellow-endosperm kernels, at various developmental stages. Following extraction from sorghum flours, carotenoids were separated by high-performance liquid chromatography (HPLC) with diode array detection. Total carotenoid content in fully matured yellow-endosperm sorghum kernels (0.112-0.315 mg/kg) was significantly lower (p < 0.05) than that in yellow maize (1.152 mg/kg) at physiological maturity. Variation in total carotenoids and within individual carotenoid species was observed in fully mature sorghum cultivars. For developing kernels, large increases in carotenoid content occurred between 10 and 30 days after half bloom (DAHB), resulting in a peak accumulation between 6.06 and 28.53 microg of total carotenoids per thousand kernels (TK). A significant (p < 0.05) decline was noted from 30 to 50 DAHB, resulting in a final carotenoid content of 2.62-15.02 microg/TK total carotenoids. (all-E)-Zeaxanthin was the most abundant carotenoid, ranging from 2.22 to 13.29 microg/TK at 30 DAHB. (all-E)-Beta-carotene was present in modest amounts (0.15-3.83 microg/TK). These data suggest the presence of genetic variation among sorghum cultivars for carotenoid accumulation in developing and mature kernels.     

Rapid electrochemical method for the evaluation of the antioxidant power of some lipophilic food extracts.

In this paper, a novel electrochemical method to evaluate the antioxidant power of lipophilic compounds present in vegetables, such as carotenoids, chlorophylls, tocopherols, and capsaicin, is reported. The method is based on a flow injection system with an electrochemical detector equipped with a glassy carbon working electrode operating amperometrically at a potential of + 0.5 V (vs Ag/AgCl). The proposed method is selective for lipophilic compounds having antioxidant power. When applied to pure compounds, the order of antioxidant power resulted as follows: lycopene > beta-carotene > zeaxanthin > alpha-carotene > beta-cryptoxanthin > lutein > alpha-tocopherol > capsaicin > chlorophyll a > chlorophyll b > astaxanthin > canthaxanthin. Results obtained on five vegetable and two fruit extracts were compared to those obtained by the 2,2'-azinobis(3-ethylbenz-thiazoline-6-sulfonic) acid (ABTS) radical cation decolorization assay, one of the most used methods to evaluate the total antioxidant capacity of foods. A good correlation between the two methods was found, except for spinach, because of the different antioxidant powers assigned by the two methods to chlorophylls. In conclusion, results suggest that the proposed electrochemical method can be successfully employed for the direct, rapid, and reliable monitoring of the antioxidant power of lipophilic food extracts.