Authentication of Rice Using Near‐Infrared Reflectance Spectroscopy

The authentication of rice (Korean domestic rice vs. foreign rice) has been attempted using near‐infrared spectroscopy (NIRS). Two sample sets (n₁ = 280 and n₂ = 200) were used to obtain calibration equations and the spectral regions used for this study were 500–600 nm, 700–900nm, and 980–2,498 nm. Modified partial least square (MPLS) regression was used to develop the prediction model. The standard error of cross validation (SECV) and the r² were 0.165 and 0.91 respectively for 1st calibration set and 0.165 and 0.93 for 2nd calibration set respectively. The results of the independent validation (n₃ = 80) showed that all of 80 samples were identified correctly. Even though authentication of rice was performed successfully using NIRS, the calibration statistics in this study showed that further effort is needed for implementation of NIRS for authentication of rice for industry purposes.     

Hardness Measurement of Bulk Wheat by Single‐Kernel Visible and Near‐Infrared Reflectance Spectroscopy

Reflectance spectra (400 to 1700 nm) of single wheat kernels collected using the Single Kernel Characterization System (SKCS) 4170 were analyzed for wheat grain hardness using partial least squares (PLS) regression. The wavelengths (650 to 700, 1100, 1200, 1380, 1450, and 1670 nm) that contributed most to the ability of the model to predict hardness were related to protein, starch, and color differences. Slightly better prediction results were observed when the 550–1690 nm region was used compared with 950–1690 nm region across all sample sizes. For the 30‐kernel mass‐averaged model, the hardness prediction for 550–1690 nm spectra resulted in a coefficient of determination (R²) = 0.91, standard error of cross validation (SECV) = 7.70, and relative predictive determinant (RPD) = 3.3, while the 950–1690 nm had R² = 0.88, SECV = 8.67, and RPD = 2.9. Average hardness of hard and soft wheat validation samples based on mass‐averaged spectra of 30 kernels was predicted and compared with the SKCS 4100 reference method (R² = 0.88). Compared with the reference SKCS hardness classification, the 30‐kernel (550–1690 nm) prediction model correctly differentiated (97%) between hard and soft wheat. Monte Carlo simulation technique coupled with the SKCS 4100 hardness classification logic was used for classifying mixed wheat samples. Compared with the reference, the prediction model correctly classified mixed samples with 72–100% accuracy. Results confirmed the potential of using visible and near‐infrared reflectance spectroscopy of whole single kernels of wheat as a rapid and nondestructive measurement of bulk wheat grain hardness.     

Comparison of Near‐Infrared Reflectance Spectroscopy and Texture Analyzer for Measuring Wheat Bread Changes in Storage

Bread staling affects bread texture properties and is one of the most common problems in bread storage. Bread firmness, as measured in compression mode by a texture analyzer (TA) has been commonly used to measure bread staling. This study investigated the potential of visible and near‐infrared reflectance spectroscopy (NIRS) to detect bread changes during storage by comparing NIRS results with those obtained by TA. Twenty‐five loaves of commercial wheat white pan bread from one batch were studied over five days. NIRS and TA measurements were made on the same slice at approximately the same time. The experiment was repeated five times using the same kind of commercial samples from five different batches. NIRS measurements of slices, loaf averages, and daily averages were compared with TA measurements. NIRS spectra had a high correlation to TA firmness. NIRS measurements correlated better with the actual storage time and had smaller standard deviations than the TA measurements. The batch differences had less effect on NIRS measurements than on the TA measurements. The results indicate that NIRS could follow bread changes during storage more accurately than the TA. NIRS is probably based on both physical and chemical changes during bread staling, unlike the TA method that only measures bread firmness, which is only one aspect of the staling phenomenon.     

Detecting and Segregating Black Tip‐Damaged Wheat Kernels Using Visible and Near‐Infrared Spectroscopy

Detection of individual wheat kernels with black tip symptom (BTS) and black tip damage (BTD) was demonstrated with near‐infrared reflectance spectroscopy (NIRS) and silicon light‐emitting‐diode (LED) based instruments. The two instruments tested, a single‐kernel NIRS instrument (SKNIRS) and a silicon LED‐based single‐kernel high‐speed sorter (SiLED‐SKS) were both developed by the Stored Product Insect and Engineering Research Unit, Center for Grain and Animal Health Research, USDA Agricultural Research Service. BTD was classified into four levels for the study ranging from sound, symptomatic (BTS) at two levels, and damaged (BTD). Discriminant analysis models for the SKNIRS instrument could distinguish sound undamaged kernels well, correctly classifying kernels 80% of the time. Damaged kernels were classified with 67% accuracy and symptomatic kernels at about 44%. Higher classification accuracy (81–87%) was obtained by creating only two groupings: 1) combined sound and lightly symptomatic kernels and 2) combined heavily symptomatic and damaged kernels. A linear regression model was developed from the SiLED‐SKS sorted fractions to predict the percentage of combined BTS and BTD kernels in a sample. The model had an R² of 0.64 and a standard error of prediction of 7.4%, showing it had some measurement ability for BTS and BTD. The SiLED‐SKS correctly classified and sorted out 90% of BTD and 66% of BTS for all 28 samples after three passes through the sorter. These instruments can serve as important tools for plant breeders and grading facilities of the wheat industry that require timely and objective determination and sorting of different levels of black tip present in wheat samples.     

Determining Vitreous Subclasses of Hard Red Spring Wheat Using Visible/Near‐Infrared Spectroscopy

The percentage of dark hard vitreous (DHV) kernels in hard red spring wheat is an important grading factor that is associated with protein content, kernel hardness, milling properties, and baking quality. The current visual method of determining DHV and non‐DHV (NDHV) wheat kernels is time‐consuming, tedious, and subject to large errors. The objective of this research was to classify DHV and NDHV wheat kernels, including kernels that were checked, cracked, sprouted, or bleached using visible/near‐infrared (Vis/NIR) spectroscopy. Spectra from single DHV and NDHV kernels were collected using a diode‐array NIR spectrometer. The dorsal and crease sides of the kernels were viewed. Three wavelength regions, 500–750 nm, 750–1,700 nm, and 500–1700 nm were compared. Spectra were analyzed by using partial least squares (PLS) regression. Results suggest that the major contributors to classifying DHV and NDHV kernels are light scattering, protein content, kernel hardness, starch content, and kernel color effects on the absorption spectrum. Bleached kernels were the most difficult to classify because of high lightness values. The sample set with bleached kernels yielded lower classification accuracies of 91.1–97.1% compared with 97.5–100% for the sample set without bleached kernels. More than 75% of misclassified kernels were bleached. For sample sets without bleached kernels, the classification models that included the dorsal side gave the highest classification accuracies (99.6–100%) for the testing sample set. Wavelengths in both the Vis/NIR regions or the NIR region alone yielded better classification accuracies than those in the visible region only.     

Selecting and Sorting Waxy Wheat Kernels Using Near‐Infrared Spectroscopy

An automated single kernel near‐infrared (NIR) sorting system was used to separate single wheat (Triticum aestivum L.) kernels with amylose‐free (waxy) starch from reduced‐amylose (partial waxy) or wild‐type wheat kernels. Waxy kernels of hexaploid wheat are null for the granule‐bound starch synthase alleles at all three Wx gene loci; partial waxy kernels have at least one null and one functional allele. Wild‐type kernels have three functional alleles. Our results demonstrate that automated single kernel NIR technology can be used to select waxy kernels from segregating breeding lines or to purify advanced breeding lines for the low‐amylose kernel trait. Calibrations based on either amylose content or the waxy trait performed similarly. Also, a calibration developed using the amylose content of waxy, partial waxy, and wild‐type durum (T. turgidum L. var durum) wheat enabled adequate sorting for hard red winter and hard red spring wheat with no modifications. Regression coefficients indicated that absorption by starch in the NIR region contributed to the classification models. Single kernel NIR technology offers significant benefits to breeding programs that are developing wheat with amylose‐free starches.     

Predicting Wheat Quality Characteristics and Functionality Using Near‐Infrared Spectroscopy

The accuracy of using near‐infrared spectroscopy (NIRS) for predicting 186 grain, milling, flour, dough, and breadmaking quality parameters of 100 hard red winter (HRW) and 98 hard red spring (HRS) wheat and flour samples was evaluated. NIRS shows the potential for predicting protein content, moisture content, and flour color b* values with accuracies suitable for process control (R² > 0.97). Many other parameters were predicted with accuracies suitable for rough screening including test weight, average single kernel diameter and moisture content, SDS sedimentation volume, color a* values, total gluten content, mixograph, farinograph, and alveograph parameters, loaf volume, specific loaf volume, baking water absorption and mix time, gliadin and glutenin content, flour particle size, and the percentage of dark hard and vitreous kernels. Similar results were seen when analyzing data from either HRW or HRS wheat, and when predicting quality using spectra from either grain or flour. However, many attributes were correlated to protein content and this relationship influenced classification accuracies. When the influence of protein content was removed from the analyses, the only factors that could be predicted by NIRS with R² > 0.70 were moisture content, test weight, flour color, free lipids, flour particle size, and the percentage of dark hard and vitreous kernels. Thus, NIRS can be used to predict many grain quality and functionality traits, but mainly because of the high correlations of these traits to protein content.     

Protein Content of Bulk Wheat from Near‐Infrared Reflectance of Individual Kernels

Protein content of wheat by near‐infrared (NIR) reflectance of bulk samples is routinely practiced. New instrumentation that permits automated NIR analysis of individual kernels is now available, with the potential for rapid NIR‐based determinations of color, disease, and protein content, all on a single kernel (sk) basis. In the event that the protein content of the bulk sample is needed rather than that of the individual kernels, the present study examines the feasibility of estimating bulk sample protein from sk spectral readings. On the basis of 318 wheat samples of 10 kernels per sample, encompassing five U.S. wheat classes, the study demonstrates that with as few as 300 kernels bulk sample protein content may be estimated by sk NIR reflectance spectra at an accuracy equivalent to conventional bulk kernel NIR instrumentation.     

Rapid Prediction of Rice Quality Characteristics by Near‐Infrared Reflectance Spectroscopy for Breeding Programs

Breeding of high‐quality rice requires quick methods to evaluate the quality characteristics such as milling, grain appearance, nutritional, eating, and cooking qualities. Because routine measurements of these quality traits are time consuming and expensive, a rapid predictive method based on near‐infrared spectroscopy (NIRS) can be applied to measure these quality parameters. In this study, calibration models for measurement of grain quality were developed using a total of 570 brown and milled rice samples. The results indicated that the models developed from the spectra of brown rice for all the quality traits had the coefficient of determination for external validation (R²) larger than 0.64 except for gel consistency. The best model was developed for the protein content, with R² of 0.94 for external validation. The model for the total score of physicochemical characteristics (TSPC), a comprehensive index reflecting all other traits, had R² of 0.70 and SD/SEP of 1.70, which indicates that high or low TSPC for a given rice could be discriminated by NIRS. The models developed from brown rice were as accurate as those from milled rice. Results suggest that NIRS‐based predictions for rice quality traits may be used as indicator traits to improve rice quality in breeding programs.     

Differentiating Vitreous and Nonvitreous Durum Wheat Kernels by Using Near‐Infrared Spectroscopy

The vitreousnss of durum wheat is used by the wheat industry as an indicator of milling and cooking quality. The current visual method of determining vitreousness is subjective, and classification results between inspectors and countries vary widely. Thus, the use of near‐infrared (NIR) spectroscopy to objectively classify vitreous and nonvitreous single kernels was investigated. Results showed that classification of obviously vitreous or nonvitreous kernels by the NIR procedure agreed almost perfectly with inspector classifications. However, when difficult‐to‐classify vitreous and nonvitreous kernels were included in the analysis, the NIR procedure agreed with inspectors on only 75% of kernels. While the classification of difficult kernels by NIR spectroscopy did not match well with inspector classifications, this NIR procedure quantifies vitreousness and thus may provide an objective classification means that could reduce inspector‐to‐inspector variability. Classifications appear to be due, at least in part, to scattering effects and to starch and protein differences between vitreous and nonvitreous kernels.     

Discriminating Oat and Groat Kernels from Other Grains Using Near‐Infrared Spectroscopy

Oats and groats can be discriminated from other grains such as barley, wheat, rye, and triticale (nonoats) with near‐infrared spectroscopy. The two instruments tested herein were the manual version of the United States Department of Agriculture–Agricultural Research Service single‐kernel near‐infrared (SKNIR) instrument and the automated QualySense QSorter Explorer high‐speed sorter, both used in similar near‐infrared spectral ranges. Three linear discriminate self‐prediction models were developed: 1) oats versus groats + nonoats, 2) oats + groats versus nonoats, and 3) groats versus nonoats. For all three models, the SKNIR instrument showed high correct classification of oats or groats (94.5–100%), which was similar to results of the QSorter Explorer at 95.0–99.4%. The amount of nonoats that were misclassified as oats or groats was low for both instruments at 0–0.2% for the SKNIR instrument and 0.8–3.7% for the QSorter Explorer. Linear discriminate models from independent prediction and validation sets yielded classification accuracies of 91.6–99.3% (SKNIR) and 90.5–97.8% (QSorter Explorer). Small differences in classification accuracy were attributed to processing speeds between the two instruments: 3 kernels/s for the SKNIR instrument and 35 kernels/s for the QSorter Explorer. This indicated that both instruments are useful for quantifying grain sample compositions of oat and groat samples and that both could be useful tools for meeting consumer demand for gluten‐free or low‐gluten products. Discrimination between grains will help producers and manufacturers meet various regulatory requirements. Examples include requirements such as those from the U.S. Food and Drug Administration and the Commission of European Communities, in which gluten‐free oats or other products can only be labeled as nongluten if they contain gluten at less than 20 ppm, the established safe consumption limit for people with celiac disease. The QSorter Explorer is currently being used to meet these requirements.     

High‐Throughput Near‐Infrared Reflectance Spectroscopy for Predicting Quantitative and Qualitative Composition Phenotypes of Individual Maize Kernels

Near‐infrared reflectance (NIR) spectroscopy can be used for fast and reliable prediction of organic compounds in complex biological samples. We used a recently developed NIR spectroscopy instrument to predict starch, protein, oil, and weight of individual maize (Zea mays) seeds. The starch, protein, and oil calibrations have reliability equal or better to bulk grain NIR analyzers. We also show that the instrument can differentiate quantitative and qualitative seed composition mutants from normal siblings without a specific calibration for the constituent affected. The analyzer does not require a specific kernel orientation to predict composition or to differentiate mutants. The instrument collects a seed weight and a spectrum in 4–6 sec and can collect NIR data alone at a 20‐fold faster rate. The spectra are acquired while the kernel falls through a glass tube illuminated with broad spectrum light. These results show significant improvements over prior single‐kernel NIR systems, making this instrument a practical tool to collect quantitative seed phenotypes at high throughput. This technology has multiple applications for studying the genetic and physiological influences on seed traits.     

Environmental Effects on Developing Wheat as Sensed by Near‐Infrared Reflectance of Mature Grains

For 30 years, near‐infrared (NIR) spectroscopy has routinely been applied to the cereal grains for the purpose of rapidly measuring concentrations of constituents such as protein and moisture. The research described herein examined the ability of NIR reflectance spectroscopy on harvested wheat to determine weather‐related, quality‐determining properties that occurred during plant development. Twenty commercial cultivars or advanced breeding lines of hard red winter and hard white wheat (Triticum aestivum L.) were grown in 10 geographical locations under prevailing natural conditions of the U.S. Great Plains. Diffuse reflectance spectra (1,100–2,498 nm) of ground wheat from these samples were modeled by partial least squares one (PLS1) and multiple linear regression algorithms for the following properties: SDS sedimentation volume, amount of time during grain fill in which the temperature or relative humidity exceeded or was less than a threshold level (i.e., >30, >32, >35, <24°C; >80%, <40% rh). Rainfall values associated with four pre‐ and post‐planting stages also were examined heuristically by PLS2 analysis. Partial correlation analysis was used to statistically remove the contribution of protein content from the quantitative NIR models. PLS1 models of 9–11 factors on scatter‐corrected and (second order) derivatized spectra produced models whose dimensionless error (RPD, ratio of standard deviation of the property in a test set to the model standard error for that property) ranged from 2.0 to 3.3. Multiple linear regression models, involving the sum of four second‐derivative terms with coefficients, produced models of slightly higher error compared with PLS models. For both modeling approaches, partial correlation analysis demonstrated that model success extends beyond an intercorrelation between property and protein content, a constituent that is well‐modeled by NIR spectroscopy. With refinement, these types of NIR models may have the potential to provide grain handlers, millers, and bakers a tool for identifying the cultural environment under which the purchased grain was produced.     

Near‐Infrared Spectroscopy for Determination of Protein and Amylose in Rice Flour Through Use of Derivatives

The use of the derivative method for near‐infrared (NIR) calibration was investigated to determine protein and amylose content in rice flour. Samples for two years, 1996 and 1999, were combined to give a wide range of the constituents for development of the calibration model. The NIR spectral data were transformed with Savitzky‐Golay derivative with multiplicative scatter correction. To develop the best derivative models, the polynomial fits (quadratic, cubic, and quartic), convolution intervals (3–11 points for protein, 3–17 points for amylose), and derivative orders (1st derivative D1; 2nd derivative D2) were investigated. For the protein analysis, all polynomial fits with 3–11 points were acceptable to develop both the D1 and D2 models. However, the three‐point quadratic and five‐point quartic fits were not acceptable for the D1 model, and the three‐point quadratic fit was not acceptable for D2. For the amylose analysis, the D1 model produced generally better results than D2. Higher convolution intervals were required for the D2 model, whereas the D1 model was not affected by convolution intervals. A quadratic (or cubic) fit with 17‐point convolution interval was acceptable for the amylose D2 model, and the quadratic fit with 5–11 points and cubic (or quartic) fit with 7–17 points were suitable for the D1 model. Based on the standard error of cross‐validation (SECV), the calibration models developed using data for two years resulted in good precision with an SECV of 0.23% for protein using four factors and an SECV of 1.0% for amylose using 10 factors.     

Prediction of Triticale Grain Quality Properties,Based on Both Chemical and Indirectly Measured Reference Methods,Using Near‐Infrared Spectroscopy

The increasing demand for triticale as food, feed, and fuel has resulted in the availability of cultivars with different grain quality characteristics. Analyses of triticale composition can ensure that the most appropriate cultivars are obtained and used for the most suitable applications. Near‐infrared (NIR) spectroscopy is often used for rapid measurements during quality control and has consequently been investigated as a method for the measurement of protein, moisture, and ash contents, as well as kernel hardness (particle size index [PSI]) and sodium dodecyl sulfate (SDS) sedimentation from both whole grain and ground triticale samples. NIR spectroscopy prediction models calculated using ground samples were generally superior to whole grain models. Protein content was the most effectively modeled quality property; the best ground grain calibration had a ratio of the standard error of test set validation to the standard deviation of the reference data of the test set (RPD_test) of 4.81, standard error of prediction (SEP) of 0.52% (w/w), and r² of 0.95. Whole grain protein calibrations were less accurate, with optimum RPD_test of 3.54, SEP of 0.67% (w/w), and r² of 0.92. NIR spectroscopy calibrations based on direct chemical reference measurements (protein and moisture contents) were better than those based on indirect measurements (PSI, ash content, and SDS sedimentation). Calibrations based on indirect measurements would, however, still be useful to identify extreme samples.     

Optimization of Near‐Infrared Reflectance Model in Measuring Gelatinization Characteristics of Rice Flour with a Rapid Viscosity Analyzer (RVA) and Differential Scanning Calorimeter (DSC)

This study compared the calibration models generated by combinations of different mathematical and preprocessing treatments as well as regression algorithms to optimize the analysis of gelatinization properties of rice flour by using near‐infrared spectroscopy, in comparison with conventional techniques of differential scanning calorimetry (DSC) and rapid viscosity analysis (RVA). A total of 220 milled rice flours were used for model construction. A model generated by the modified partial least squares regression (MPLS) with mathematical treatment “2, 8, 8, 2” (second‐order derivative computed based on eight data points, and eight and two data points in the second smoothing, respectively) and detrend preprocessing was identified as the best for simultaneously measuring onset temperature (T_o), peak temperature (T_p), and conclusion temperature (T_c) of DSC. MPLS/“2, 8, 8, 2”/weighted multiplicative scattering correction preprocessing was identified as the best for RVA properties. The results indicated that near‐infrared reflectance spectroscopy could be used to rapidly predict gelatinization properties of rice flour for the purposes of quality evaluation of germplasm and selection of intermediate lines in breeding programs.     

Classification of Single‐ and Double‐Mutant Corn Endosperm Genotypes by Near‐Infrared Transmittance Spectroscopy

A total of 1,176 grain samples representing 10 different single‐ and double‐mutant genotypic classes of specialty starch corn were used for developing various classification models based on near‐infrared transmittance spectra. The genotypes used included amylose‐extender (ae), dull (du), sugary‐2 (su2), waxy (wx), ae wx, ae du, ae su2, du wx and du su2. Two‐class classification models (only two genotypes compared) were developed using partial least squares analysis (PLS) while three‐way and multiclass models were examined using principal component analysis (PCA). The effectiveness of the calibrations was evaluated by examining the percentage of unknown grain samples incorrectly classified. In general, two‐class models performed better than multiclass models. However, they did not show improvement when discriminating among genotypes with overlapping amylose contents such ae du vs. ae and ae su2 vs. ae. Three‐way models including double‐mutants and their corresponding single‐mutant counterparts had misclassification percentages typically <5% using 14 PCA factors but again, with the exception of models including genotypes with overlapping amylose contents such as ae du vs. ae vs. du. The best multiclass model using all 10 genotypic classes simultaneously revealed only two classes (ae su2 and du) with misclassification rates >10% based on 16 PCA factors. This study demonstrates that, depending on the material to be considered, near‐infrared transmittance spectroscopy could be useful when segregation of specialty starch hybrids grain from other grain types is necessary.     

Staling Effects When Adding Low Amounts of Normal and Heat‐Treated Barley Flour to a Wheat Bread

The properties of a white wheat bread could be changed by adding normal or heat‐treated barley flour in small amounts (2 and 4%) to a white wheat bread recipe. Differences regarding gelatinization as well as retrogradation properties were found when analyzing the two flours in model systems. The heat‐treated flour was fully gelatinized due to prior time, temperature, and pressure treatment and could therefore absorb larger amounts of water than the other flours. In gelatinized model systems with 40% flour (dwb), the heat‐treated barley flour contained less retrograded amylopectin as compared with normal barley flour after storage for up to 14 days, whereas no differences were found with 20% flour (dwb). However, stored breads showed an increased retrogradation of amylopectin (as measured by differential scanning calorimetry [DSC]) when 2% pretreated barley flour was added as compared with addition of 2% normal barley flour. On the other hand, there were no significant differences at the 4% level. Addition of either of the barley flours resulted in less firm breads during storage as compared with the control breads. Increased water absorption in barley flour and thus increased water content in the breads or different water‐binding capacities of the flour blends could explain these results. The present study indicated that water had a stronger influence on bread firmness than the retrogradation of amylopectin. This conclusion was based on breads with pretreated barley flour being less firm than breads with normal barley flour, although the retrogradation, as determined by DSC, was higher.