Industrial Crops and Products 24 (2006) 307313

The use of near-infrared spectroscopy (NIRS) in the study of seed quality components in plant breeding programs
Rafael Font a , Mercedes del R o-Celestino b , Antonio de Haro-Bail on a,
a b

Instituto de Agricultura Sostenible (C.S.I.C.), Alameda del Obispo s/n, 14080 C ordoba, Spain CIFA-Alameda del Obispo (I.F.A.P.A.-C.I.C.E., Junta de Andaluc a), 14080 C ordoba, Spain

Abstract The potential of near-infrared spectroscopy (NIRS) for determining acid detergent ber (ADF), fatty acid composition, and also protein and oil content in the seed of oilseed Brassica (fam. Brassicaceae) and chickpea (Cicer arietinum L.) was assessed. Accessions of the species Indian mustard (Brassica juncea L. Czern. & Coss.), Ethiopian mustard (Brassica carinata A. Braun) and rapeseed (Brassica napus L.), and also chickpea (Cicer arietinum L.), were scanned by NIRS as intact (Brassica spp.) or ground (C. arietinum) seed, and their reference values regressed against different spectral transformations by modied partial least squares (PLSm) regression. The coefcients of determination in the external validation (r2 ) for the different components analyzed in Brassica ranged from 0.81 (oleic acid C18:2 in B. carinata) to 0.98 (oil in B. juncea), which characterize those equations as having from good to excellent quantitative information. The standard deviation (S.D.) to standard error of prediction ratio (RPD) and S.D. to range (RER) were variable for the different species and seed constituents, and showed values that were characteristic of equations suitable for screening purposes to select samples for more detailed chemical analysis. Also, quality components in chickpea seed were determined accurately. NIRS equations showed r2 values that ranged from 0.85 (C18:3) to 0.95 (crude protein and C22:1), and RPD and RER values that were indicative of equations suitable for screening. 2006 Elsevier B.V. All rights reserved.
Keywords: Near-infrared spectroscopy; Oilseed Brassica spp.; Cicer arietinum; Quality components; Plant breeding

1. Introduction The importance of the oilseed Brassica and legumes in human and animal nutrition, and also as crops with industrial application, is well recognized (Uppstr om, 1995). Over the past three decades the production of oilseed Brassica has increased to the state where it is one of the most important world sources of vegetable oil. Improvements in the quality of oil and meal have resulted in the recognition of the oil as an edible product of high nutritional value, and the meal as an important

Corresponding author. Tel.: +34 957499247; fax: +34 957499252. E-mail address: cs9habaa@uco.es (A. de Haro-Bail on).

source of protein for animal nutrition. Simultaneously, genotypes of Brassica exhibiting high levels of erucic acid (C22:1) in oil have been developed (Velasco et al., 1998), with applications in the lubricants industry. The utilization of the oil for fuel has also been commercialized (Cardone et al., 2003), as a substitute for traditional fossil diesel fuel, to reduce greenhouse gas emissions. Legumes have been well recognized as a valuable food in human nutrition (S anchez-Vioque et al., 1998), particularly in developing countries. Food legumes are an excellent source of dietary protein in animal feed rations and in human diets. Edible legumes have also been used as a part of the dietary treatment of diabetes (Thorne et al., 1983) and fortication of foods (Shehata et al., 1988).

0926-6690/$ see front matter 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.indcrop.2006.06.012

308

R. Font et al. / Industrial Crops and Products 24 (2006) 307313

Plant breeding programs usually involve extensive evaluations of the quality components of interest. Thus, large numbers of screenings by the standard analytical methods of seed lines are usually performed, in order to detect target genotypes. Although the standard analytical techniques usually offer a high level of accuracy and precision, they also show some handicaps, such as high costs, high labor input and delay in reporting. In addition, many standard techniques involve the destruction of the test sample, which could be a handicap in the case of valuable and scarce materials. In recent decades the development of equipment featuring improved electronic and optical components, the advent of computers capable of effectively processing information contained in spectra and development of powerful chemometric applications, have facilitated the expansion of spectroscopic techniques in an increasing number of elds, allowing efcient management of spectral and chemical data. The use of fast analytical techniques such as nearinfrared spectroscopy (NIRS) has many advantages compared with standard techniques. NIRS analysis is carried out with considerable saving of time, cost and without using hazardous chemicals. In addition, samples can be analyzed in their natural form without destruction, and simultaneous analyses of several traits may be carried out. Absorption of radiation in the region of 4002500 nm (visible plus near-infrared regions) is used to develop calibration curves, which can be related to sample properties. After calibration, the regression equations developed allows accurate analysis of many other samples by prediction of data based on the spectra. In recent decades, NIRS has been widely used as a fast and reliable method for qualitative and quantitative analysis of biological and non-biological materials in agriculture, food, textile, petrochemical, environmental and pharmaceutical elds (Williams and Norris, 1987; Murray and Cowe, 1992; Font et al., 2004a). International Standards Committees have formally accepted methods using NIRS for analysis of many compounds (Batten, 1998). Regarding Brassicas, many authors have reported NIRS equations for different components, such as glucosinolates (Tkachuk, 1981; Michalski and Krzymanski, 1988; Velasco and Becker, 1998; Font et al., 2004b), ber (Michalski et al., 1992; Font et al., 2003), protein, oil and fatty acids (Tkachuk, 1981; Font et al., 2002). In legumes, NIRS has been successfully applied to determine tannins in Vicia faba L. (De Haro et al., 1989), but it has never been used to assess the composition of other important legume species such as chickpea (Cicer arietinum L.),

which is the third most important legume species (FAO, 1993) in the world. The Department of Agronomy and Plant Breeding (Institute of Sustainable Agriculture, CSIC) for many years, has focused on applying NIRS to plant breeding programs to study multiple seed quality components of different plant species of agronomical interest, including oilseed Brassica and legumes. In this paper, we present some of the results obtained in relation to the use of NIRS to analyze protein, oil, fatty acids, acid detergent ber and glucosinolates in the genetically related allotetraploid species of Brassica Indian mustard (Brassica juncea L. Czern. & Coss.; genome = AABB), Ethiopian mustard (Brassica carinata A. Braun; genome = BBCC) and rapeseed (Brassica napus L.; genome = AACC). In addition, NIRS calibrations for crude protein, oil and fatty acid composition of C. arietinum seeds are also reported. 2. Materials and methods 2.1. Scanning samples for NIRS analysis A NIR Systems Model 6500 spectrophotometer (Foss-NIRSystems, Inc., Silver Spring, MD, USA) equipped with a transport module was used to perform the NIRS analyses. Seed samples were analyzed as intact (Brassica spp.) or ground seed (C. arietinum). Spectra were recorded in reectance mode. In this mode, a ceramic standard is placed in the radiant beam, and the diffusely reected energy is measured at each wavelength, before and after reading the sample. Spectra of the samples were recorded once from each sample, and were obtained as an average of 32 scans. The ceramic and the sample spectra were used to generate the nal log (1/R) spectrum. Seed samples were placed for the analysis in a 3 cm diameter round cell holder. This cell is composed of quartz glass and anodized aluminium to avoid absorption. Intact or ground seed samples lled the sample holder (10 ml volume, approximately) and their spectra were registered in the range from 400 to 2500 nm, at 2 nm intervals. 2.2. Selecting the samples for NIRS analysis Samples used in the NIRS analysis were selected to represent the whole spectral and chemical variability in the target population in the calibration and validation groups. This enabled principal component analysis (PCA) to be performed on the registered spectra. The Mahalanobis distance (H) was used as a criterion for

R. Font et al. / Industrial Crops and Products 24 (2006) 307313

309

selecting those samples in the population as being more variable on the basis of spectra features (Shenk and Westerhaus, 1991). The ISI algorithm CENTER was used to establish population boundaries with a maximum standardized H distance of 3.0. Then, the algorithm SELECT was used for efcient selection, by choosing samples with a minimum standardized H distance of 0.6 from their nearest neighbors. The nal number of samples selected for NIRS analysis was variable, depending on different economical and practical factors. 2.3. Obtaining the reference values of the selected samples For each of the species, the selected samples were analyzed separately for the different quality components of interest by the respective standard methods of analysis. Acid detergent ber (ADF) was determined according to procedures described by Goering and Van Soest (1970) in a Dosi-Fiber apparatus (SELECTA). Samples were analyzed in duplicate, and both determinations averaged. Glucosinolates were determined according to ISO protocol (ISO 9197-1, 1992). Values of individual desulfoglucosinolates were determined with a model 600 HPLC instrument (Waters), equipped with a model 486 UV tunable absorbance detector at a wavelength of 229 nm. Separation was carried out by using a Lichrospher 100 RP-18 in Lichrocart 125-4 column, 5 m particle size (Merck). The amount of each individual glucosinolate present in the sample was calculated by mean of the internal standard, and expressed as mol g1 seed dry weight. The total glucosinolate content was computed as the sum of all the individual glucosinolates present in the sample. Fatty acid methyl esters were obtained as described by Garc es and Mancha (1993) and analyzed on a PerkinElmer Autosystem gasliquid chromatograph (PerkinElmer Corporation, Norwalk, USA) equipped with a amed ionization detector (FID). Oil content in the seeds was determined by using nuclear magnetic resonance (NMR), according to the protocol of the AOCS (1980). Nitrogen content was determined by Kjeldahl analysis, and a factor of 6.25 was used to estimate the protein content (AOAC, 1990). 2.4. Developing NIRS equations Spectra corresponding to the samples were sorted on the basis of the reference values for each component, from the lowest to the highest, and then divided into calibration and validation groups in a rate 2:1.

In a rst step, the full wavelength range (from 400 to 2500 nm) was used for calibration. In most cases, the visible region of the spectrum (400700 nm) provided efcient contribution to calibration of seed components, as color is often correlated with seed characters (i.e., chlorophyll and pigments, protein, moisture, oil and carbohydrates) (Williams and Sobering, 1996). Calibrations were performed by using the GLOBAL v. 1.50 software (WINISI II, Infrasoft International, LLC, Port Matilda, PA, USA). Calibration equations were computed by using the raw optical data (log 1/R), or rst or second derivatives of the log 1/R data, with several combinations of segment (smoothing) and derivative (gap) sizes [i.e., (0, 0, 1, 1; derivative order, segment of the derivative, rst smooth, second smooth); (1, 4, 4, 1); (1, 10, 10, 1); (2, 5, 5, 2); (2, 10, 10, 2)] (Shenk et al., 1992). The use of derivative spectra instead of the raw optical data to perform calibration is a way of solving problems associated with overlapping peaks and baseline correction (Hruschka, 1987). A rst-order derivative of log(1/R) results in a curve containing peaks and valleys corresponding to the point of inection on either side of the log(1/R) peak. While the second-order derivative calculation results in a spectral pattern display of absorption peaks pointing down rather than up, with an apparent band resolution (Shenk et al., 1992). The gap size and amount of smoothing used to make the transformation will also affect the number of apparent absorption peaks. Among the different methods based on selected wavelengths or in full-spectrum which are available in commercial chemometric softwares, the full-spectrum methods have yield calibration equations that perform better with the types of seed material, and components that are the object of this study. Specially, modied partial least squares (PLSm) regression has been revealed as noteworthy for assessing seed components. PLS performs a linear regression in a new coordinate system with a lower dimensionality than the original space of independent variables. The PLS factors are determined by the maximum variance of independent (spectral data) variables and by a maximum correlation with the dependent (chemical) variable. The model actually uses only the primary, most important factors, the noise being encapsulated in the less important factors. Regression is performed in the space spanned by the new reduced coordinate system of the orthogonal factors. In addition to derivatives, standard normal variate and de-trending (SNVD) algorithms (Barnes et al., 1989) were applied to the derived spectra to minimize baseline offset due to scattering effects caused by differences in particle size or path-length variation among samples.

310

R. Font et al. / Industrial Crops and Products 24 (2006) 307313

2.5. Validation of the equations An external validation procedure was carried out to determine the accuracy and precision of the equations obtained in the calibration for each component in each species. To evaluate the accuracy of the equations, different statistics were used, namely the coefcient of determination (r2 ) (Williams, 1987); the RPD, which is the ratio of the standard deviation (S.D.) for the validation samples to the standard error of prediction (performance) (SEP), and the RER, which is the ratio of the range in the reference data (validation set) to the SEP (Williams and Sobering, 1996). As far as possible, we also calculated the ratio SEP to standard error of laboratory (SEL), as this statistic allows the error of NIRS to be put in perspective to the error in the reference method. The mathematical expressions of these statistics are as follow:
n n 1

N = number of samples, K = number of wavelengths used in an equation. 3. Results and discussion 3.1. Acid detergent ber Fiber is a component of high interest in Brassica spp., as it has been demonstrated to be negatively correlated with the oil and protein content in seed and with meal digestibility (Simbaya et al., 1995). Studies carried out by our research group in previous years have shown that ADF calibrations performed over single species of Brassica, yielded prediction accuracies that were not signicantly different from those obtained for multiproduct calibrations for the related allotetraploid species B. juncea, B. carinata and B. napus. Those ndings led to the development of a multi-species calibration for ADF over the above mentioned species, which is continuously expanded with new samples each time any accession showing chemical or physical variability is detected. This strategy allows a lower number of reference analyses to perform the calibration, and avoids the need to maintain separate calibrations by species. Thus, the multi-product equation obtained for ADF determination resulted in an r2 (Eq. (1)) value (0.83) in the external validation that was indicative of good quantitative information equations (Shenk and Westerhaus, 1996) (Table 1; Fig. 1). Although the highest RPD (Eq. (2)) obtained (2.40) was under the cut-off point of 3, which is recommended by Williams and Sobering (1996) for using the equation for screening purposes, the RER (Eq. (3)) displayed (10.75), is considered by the same authors as suitable for screening on the basis of the set range. The ratio SEP/SEL (Eq. (4)) obtained for the

r =
2 i=1

( yy )

2 i=1

(yi y )

(1)

where y = NIR measured value; y = mean y value for all samples; yi = lab reference value for the ith sample.
n 1/2 1

RPD = S.D.
i=1

(yi y i)

(N K1)

(2)

where yi = lab reference value for the ith sample; y = NIR measured value; N = number of samples, K = number of wavelengths used in an equation; S.D. = standard deviation.
n 1/2 1

RER = range
i=1

(yi y i)

(N K1)

(3)

where yi = lab reference value for the ith sample; y = NIR measured value; N = number of samples, K = number of wavelengths used in an equation. ratio SEP = SEL
i=1 n 1/2

(yi y i )2
i=1 n

(N K 1)1
1/2 1

(y1 y2 )

(2N )

(4)

where yi = laboratory reference value for the ith sample; y = NIR measured value; y1 and y2 = laboratory reference values for two duplicates of the same sample;

Fig. 1. External validation scatter plot for acid detergent ber (% dw) of laboratory vs. predicted data by NIRS in Brassica spp.

R. Font et al. / Industrial Crops and Products 24 (2006) 307313 Table 1 Validation statistics for equations of different components developed over the intact seed of Brassica juncea (70 < n < 300) Component Acid detergent bera (% dw) Glucosinolates (mol g1 dw) Sinigrin (mol g1 dw) Gluconapin (mol g1 dw) Oil (% dw) C18:1 (% oil) C18:2 (% oil) C18:3 (% oil) C22:1 (% oil) Range 6.6515.47 21.89187.29 4.70178.25 0.51142.56 24.4252.04 8.3450.00 11.2743.51 5.1114.02 0.0256.23 Mean 11.02 127.62 68.51 54.92 36.48 19.73 22.87 9.47 38.37 S.D. 1.97 34.15 48.58 49.66 5.73 11.75 7.62 1.78 14.66 SEP 0.82 15.65 18.74 18.12 0.82 1.93 1.51 0.75 2.54 r2 0.83 0.82 0.86 0.95 0.98 0.97 0.96 0.82 0.97 RPD 2.40 2.18 2.59 2.74 6.98 6.08 5.04 2.37 5.77

311

RER 10.75 10.56 9.26 12.09 33.68 21.58 21.35 11.88 22.12

n: Number of samples in the validation set; S.D.: standard deviation of the data in the validation set; SEP: standard error of performance; r2 : coefcient of determination in the validation; RPD: ratio of the standard deviation to standard error of prediction (performance); RER: ratio of the range to standard error of prediction (performance); dw: dry weight; C18:1 = oleic acid, C18:2 = linoleic acid, C18:3 = linolenic acid, and C22:1 = erucic acid. a Multi-product equation comprising the species B. juncea, B. carinata and B. napus.

second derivative of the equation (3.56) was characteristic of high precision equations in the prediction of ADF. 3.2. Individual and total glucosinolates Genotypes of B. juncea have been reported to show a large variability in its individual glucosinolate pattern depending on the origin of the cultivar (Uppstr om, 1995). For instance, B. juncea genotypes of European or North American origin contain high quantities of sinigrin (typically shows values ranging from 150 to 200 mol g1 , oil-extracted, air dried meal), while genotypes from the Indian subcontinent contain variable amounts of sinigrin and gluconapin. In contrast, sinigrin is the major glucosinolate contained in the B. carinata seed, which shows low concentrations of other minor glucosinolates. However, these compounds can be measured by NIRS, as individual glucosinolates (sinigrin, gluconapin) or as total glucosinolates, in spite of the low concentrations in which they are usually present in the seeds (Table 1). In developing NIRS equations for sinigrin and gluconapin in B. juncea seeds, the second derivative transformation (2, 5, 5, 2; SNV + DT) equation of the raw (log 1/R) data resulted in the highest accuracy

of any of the other mathematical treatments that were tested. The r2 values shown by the sinigrin (0.86) and by the gluconapin (0.95) equations were indicative of equations showing good and excellent quantitative information, respectively, i.e., the 86 and 95% of the sinigrin and gluconapin variability contained in the seed samples of their respective validation sets were explained by the models. The RPD (2.59; 2.74) and RER (9.26; 12.09) values obtained for sinigrin and gluconapin, respectively, were close to those values recommended for screening purposes (Williams and Sobering, 1996). Moreover, the equation for total glucosinolates also showed good quantitative information (r2 = 0.82) and the RPD and RER values were suitable for screening such components. To evaluate the prediction ability of those equations in relation to the overall error of the reference method, the SEL was calculated and related to SEP. The SEP/SEL ratio shown by sinigrin and gluconapin (2.70 and 2.08, respectively) and by the total glucosinolate (1.60) equations, classied them as having good (for sinigrin and gluconapin) or excellent (total glucosinolates) precisions. The NIRS equation showing the highest prediction ability for determining total glucosinolates over B. cari-

Table 2 Validation statistics for equations of different components developed over the intact seed of Brassica carinata (100 < n< 180) Component Crude protein (% dw) Glucosinolates (mol g1 dw) Oil (% dw) C18:1 (% oil) C18:2 (% oil) C18:3 (% oil) C22:1 (% oil) Range 16.8037.80 56.53221.10 25.6054.60 4.5365.75 7.1533.46 3.1121.18 0.5635.00 Mean 24.23 117.62 43.12 14.38 17.17 12.28 38.26 S.D. 4.66 31.65 5.38 10.51 4.67 2.99 12.25 SEP 0.51 8.50 1.49 3.77 2.04 1.17 2.81 r2 0.98 0.93 0.92 0.90 0.81 0.85 0.95 RPD 6.85 3.72 3.61 2.78 2.28 2.55 4.35 RER 41.17 19.36 21.48 16.23 12.89 15.44 12.25

312

R. Font et al. / Industrial Crops and Products 24 (2006) 307313

Table 3 Validation statistics for equations of different components developed over the ground seed of Cicer arietinum (30 < n < 50) Component Crude protein (% dw) Oil (% dw) C18:1 (% oil) C18:2 (% oil) Range 16.6627.50 2.475.77 19.5639.81 46.4764.40 Mean 22.78 4.23 26.37 57.79 S.D. 2.53 0.75 4.27 3.91 SEP 0.55 0.25 1.32 1.16 r2 0.95 0.91 0.90 0.91 RPD 4.60 3.00 3.23 3.37 RER 19.70 13.20 15.34 15.45

nata seed (Table 2), showed slightly higher accuracy than those for B. juncea. The high r2 (0.93) obtained in the validation and, the RPD (3.72) and RER (19.36) values indicated an equation useful for screening purposes. 3.3. Oil and fatty acids Most oil and fatty acid equations developed for Brassica spp. seed showed sufcient accuracy for using this technique as a valuable tool for the analysis of these components (Tables 1 and 2). The r2 shown by the equations for oil and fatty acid determination in B. juncea seeds, indicated from good to excellent quantitative information (Shenk and Westerhaus, 1996). On the other hand, on the basis of the statistics RPD and RER, most of the equations were higher than 3 and 10, respectively, thus being useful for screening (Williams and Sobering, 1996). The inclusion in the NIRS analysis of accessions showing a high variability could explain the increment observed by the SEP (Table 2), and therefore, the lower RPDs obtained for B. carinata in the equations for oil and fatty acids in contrast to those of B. juncea. However, the RERs obtained for these components, were higher than the minimum value recommended for screening purposes, and the r2 were, for all the equations, indicative of good or excellent quantitative information. NIRS also determined oil (Fig. 2) and fatty acids accurately in chickpea. Equations for these components showed similar performances in the validation (Table 3), which exhibited r2 values of 0.90 for oleic acid (C18:1) and 0.91 for oil and linoleic acid (C18:2). These results characterized those equations as having excellent quantitative information (Shenk and Westerhaus, 1996). In addition, the RPD and RER ratios obtained were above 3 and 13, respectively, thus, making these equations suitable for screening purposes (Williams and Sobering, 1996). 3.4. Crude protein Crude protein in B. carinata was predicted by NIRS with a high accuracy (Table 3). The r2 , RPD and RER

Fig. 2. External validation scatter plot for oil (% dw) of laboratory vs. predicted data by NIRS in Cicer arietinum L.

shown by this equation, together with those for oil in B. juncea, were the highest of all components studied. These statistics verify accurate analysis for this component (Williams, 1987). Crude protein was also accurately determined by NIRS in C. arietinum (Table 3). The high r2 (0.95) shown, and also the RPD and RER statistics, indicated a high prediction ability equation. 4. Conclusion The use of the CENTER and SELECT algorithms has allowed efcient selection of samples for NIRS analysis of the different seed components studied. Thus, by using these mathematical tools sufcient representation of the whole population by those samples selected for wet chemistry analyses was ensured, which implied an important reduction of laboratory input and operation cost associated with screening. The NIRS equations developed over Brassica spp. and Cicer arietinum seeds have allowed accurate predictions of multiple components in a simultaneous way. Thousands of seed samples are generated by plant selection and breeding programs which commonly look to develop progress in several traits simultaneously. Low cost, rapid methods are required to support such pro-

R. Font et al. / Industrial Crops and Products 24 (2006) 307313

313

grams. Reectance spectroscopy makes NIRS an ideal tool for screening traits in plant breeding programs. Acknowledgements Authors are grateful to Dr. R. Garc a Ru z, Department of Plant Breeding and Agronomy, CIFA (Junta de Andaluc a, C ordoba) for providing the samples of Brassica napus used in this study, and also to Eileen Brophy for the grammatical revision of this text. This work has been supported by Project CICYT no. AGL 2001-2293 of the Spanish Government. References
AOAC, 1990. Ofcial Methods of Analysis. Association of Ofcial Analytical Chemists, Arlington, USA. AOCS, 1980. Ofcial Methods and Recommended Practices of the American Oil Chemists Society. Champaign, IL, USA. Barnes, R.J., Dhanoa, M.S., Lister, S.J., 1989. Standard normal variate transformation and de-trending of near-infrared diffuse reectance spectra. Appl. Spectrosc. 43, 772777. Batten, G.D., 1998. Plant analysis using near infrared reectance spectroscopy: the potential and the limitations. Aust. J. Exp. Agr. 38, 697706. Cardone, M., Mazzoncini, M., Menini, S., Rocco, V., Senatore, A., Seggiani, M., Vitolo, S., 2003. Brassica carinata as an alternative oil crop for the production of biodiesel in Italy: agronomic evaluation, fuel production by transesterication and characterization. Biomass Bioenerg. 25, 623636. De Haro, A., L opez-Medina, J., Cabrera, A., Mart n, A., 1989. Determination of tannin in the seeds of Vicia faba by NIR. In: Huisman, J., van der Poel, T.F.B., Liener, I.E. (Eds.), Recent Advances of Research in Antinutritional Factors in Legume Seeds. Pudoc, Wageningen, pp. 297300. FAO, 1993. FAO Yearbook. Production 1992. FAO. Font, R., del R o, M., Sillero, A., Arthur, E., Bancroft, I., Chinoy, C., Morgan, C., De Haro, A., 2002. Using near infrared spectroscopy for determining protein content in Ethiopian mustard (Brassica carinata A Braun). Cruciferae Newslett. 24, 78. Font, R., del R o, M., Fern andez, J.M., de Haro, A., 2003. Acid detergent ber analysis in oilseed Brassicas by near-infrared spectroscopy. J. Agric. Food Chem. 51, 29172922. Font, R., del R o, M., V elez, D., Montoro, R., de Haro, A., 2004a. Use of near-infrared spectroscopy for determining the total arsenic content in prostrate amaranth. Sci. Total Environ. 327, 93104. Font, R., del R o, M., Fern andez-Mart nez, J.M., de Haro-Bail on, A., 2004b. Use of near-infrared spectroscopy for screening the individual and total glucosinolate contents in Indian mustard seed (Brassica juncea L Czern. & Coss.). J. Agric. Food Chem. 52, 35633569. Garc es, R., Mancha, M., 1993. One step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Anal. Biochem. 211, 139143. Goering, H.K., Van Soest, P.J., 1970. Forage ber analysis: apparatus, reagents, procedures, and some applications. USDA-ARS Agric. Handbook No. 379. U.S. Gov. Print. Ofce, Washington, DC. Hruschka, W.R., 1987. Data analysis: wavelength selection methods. In: Williams, P., Norris, K. (Eds.), Near-infrared Technology in the

Agricultural and Food Industries. American Association of Cereal Chemists, Inc., pp. 3555. ISO norm, 1992. RapessedDetermination of glucosinolates contentPart 1: Method using high-performance liquid chromatography. ISO 9167-1, 19. Michalski, K., Krzymanski, J., 1988. Application of NIR method for screening of oilseed rape on glucosinolate content. Cruciferae Newslett. 13, 5455. Michalski, K., Ochodzki, P., Cicha, B., 1992. Determination of bre, sulphur amino acids and lysine in oilseed rape by NIT. In: Murray, I., Cowe, I.A. (Eds.), Making Light Work: Advances in Near Infrared Spectroscopy. VCH Verlagsgesellschaft, Weinheim (Federal Republic of Germany) and VCH Publishers, NY, pp. 333335. Murray, I., Cowe, I.A., 1992. Making Light Work: Advances in Near Infrared Spectroscopy. VCH Verlagsgesellschaft, Weinheim and VCH Publishers, NY. S anchez-Vioque, R., Clemente, A., Vioque, J., Bautista, J., Mill an, F., 1998. Polar lipids of defatted chickpea (Cicer arietinum L.) our and protein isolates. Food Chem. 63, 357361. Shehata, N.A., Darwish, N., El-Nahry, X., Rarack, F.A.A., 1988. Supplementation of wheat our with some local legumes. Die Naharung 32, 310. Shenk, J.S., Westerhaus, M.O., 1991. Population structuring of near infrared spectra and modied partial least squares regression. Crop Sci. 31, 15481555. Shenk, J.S., Workman Jr., J.J., Westerhaus, M.O., 1992. Application of NIR spectroscopy to agricultural products. In: Burns, D.A., Ciurczak, E.W. (Eds.), Handbook of Near-Infrared Analysis. Dekker Inc., NY, pp. 383431. Shenk, J.S., Westerhaus, M.O., 1996. Calibration the ISI way. In: Davies, A.M.C., Williams, P.C. (Eds.), Near Infrared Spectroscopy: The Future Waves. NIR Publications, Chichester, pp. 198202. Simbaya, J., Slominski, B.A., Bogdan, A., Rakow, G., Campbell, L.D., Downey, R.K., Bell, J.M., 1995. Quality characteristics of yellowseeded Brassica seed meals: protein, carbohydrates, and dietary ber components. J. Agric. Food Chem. 43, 20622066. Thorne, M.J., Thompson, L.U., Jenkins, D.J.A., 1983. Factors affecting starch digestibility and the glycemic response with special reference to legumes. Am. J. Clin. Nutr. 38, 481485. Tkachuk, R., 1981. Oil and protein analysis of whole rapeseed kernels by near infrared reectance spectroscopy. J. Am. Oil Chem. Soc. 58, 819822. Uppstr om, B., 1995. Seed chemistry. In: Kimber, D., McGregor, D.I. (Eds.), Brassica Oilseeds. Production and Utilization. CAB International, Wallingford, pp. 217242. Velasco, L., Fern andez Mart nez, J.M., De Haro, A., 1998. Increasing erucic acid content in Ethiopian mustard through mutation breeding. Plant Breed. 117, 8587. Velasco, L., Becker, H.C., 1998. Analysis of total glucosinolate content and individual glucosinolates in Brassica spp. by near-infrared reectance spectroscopy. Plant Breed. 117, 97102. Williams, P.C., 1987. Variables affecting near-infrared reectance spectroscopy analysis. In: Williams, P., Norris, K. (Eds.), Nearinfrared Technology in the Agricultural and Food Industries. American Association of Cereal Chemists, Inc., pp. 143167. Williams, P.C., Norris, K., 1987. Near-Infrared Technology in the Agricultural and Food Industries. AACC, Inc., USA. Williams, P.C., Sobering, D.C., 1996. How do we do it: a brief summary of the methods we use in developing near infrared calibrations. In: Davies, A.M.C., Williams, P.C. (Eds.), Near Infrared Spectroscopy: The Future Waves. NIR Publications, Chichester, pp. 185188.