Sens. & Instrumen. Food Qual. (2009) 3:219226 DOI 10.

1007/s11694-009-9088-y

ORIGINAL PAPER

Prediction of white button mushroom (Agaricus bisporus) moisture content using hyperspectral imaging
Masoud Taghizadeh Aoife Gowen Colm P. ODonnell

Received: 22 May 2009 / Accepted: 9 October 2009 / Published online: 25 October 2009 Springer Science+Business Media, LLC 2009

Abstract Hyperspectral imaging is a non-contact, nondestructive technique that combines spectroscopy and imaging to extract information from a sample. This technology has recently emerged as a powerful technique for food analysis. In this study, the potential of hyperspectral imaging (HSI) to predict white button mushroom moisture content (MC) was investigated. Mushrooms were subjected to dehydration at 45 1 C for different time periods (0, 30, 60 and 120 min) to obtain representative samples at different moisture levels (93.40 0.62%, 82.76 2.11%, 73.20 2.60% and 60.89 4.32% wet basis [wb]). Hyperspectral images of the mushrooms were obtained using a pushbroom system operating in the wavelength range of 4001000 nm. Hunter L, a and b colour values of the mushrooms were also measured. The average reectance spectra of samples at different MC levels were obtained and Partial Least Square Regression (PLSR) models were built to predict mushroom moisture content. To reduce the spectral variability caused by factors unrelated to MC such as scattering effects and differences in sample height, different spectral pre-treatments were applied. The Standard Normal Variate (SNV) transformation was found to be the best approach among the wavelength range studied, resulting in the greatest reduction in Root Mean Square Error of Cross Validation (RMSECV) and Root Mean Square Error of Prediction (RMSEP) for a 4-component PLSR model. RMSECV of 5.50 (% wb) and RMSEP of 5.58 (% wb) were obtained for the calibration and test sets of data, respectively. Prediction maps were
M. Taghizadeh (&) A. Gowen C. P. ODonnell Biosystems Engineering, School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Beleld, Dublin 4, Ireland e-mail: masoud.taghiza@ucd.ie

generated from hyperspectral data to show the predictive model performance at pixel level. This study shows the potential of hyperspectral imaging for prediction of mushroom moisture content in the studied wavelength range. The implemented method highlighted contrast between areas of different moisture content to achieve better knowledge of dehydration distribution over the mushroom surface. Keywords Hyperspectral imaging Mushroom Moisture content Partial least square regression

Introduction White button mushroom is an edible fungus widely used in the food industry. It is the main commercial variety of fungi utilised for processing in many countries, including China, the United States and some countries in Europe [1]. Short shelf life and high product variability are some of the most challenging problems in the mushroom industry. Recent research in the area of shelf life of mushrooms mainly focused on shelf life extension by use of different post harvest treatments and innovative packaging design [24]. One of the most important parameters affecting mushroom quality and shelf life is moisture content [5]. Measurement of moisture content (MC) is also very important in the production of dried mushroom. Roy et al. [6] used VisNIR spectroscopy in the wavelength range (6002200 nm) for non-destructive prediction of the moisture content of fresh mushrooms. Hyperspectral imaging (HSI) is a novel technique that combines conventional imaging and spectroscopy to acquire both spatial and spectral information from an object. It can also be used to extract some intrinsic

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chemical and molecular information from a product. In order to acquire a hyperspectral image a three dimensional image cube, or hypercube, is constructed which contains two spatial and one wavelength dimension [7]. This technique was initially developed for remote sensing applications but has since found applications in different elds such as astronomy, pharmaceuticals, food and medicine [817]. The main advantages of hyperspectral imaging are: minimal sample preparation, simultaneous determination of several components, estimation of component concentration and distribution within a sample. Much research has been carried out investigating the potential application of hyperspectral imaging in monitoring the quality and safety attributes of a variety of agricultural food products such as fruits, vegetables, meat, poultry and grains [1821]. A limited number of studies have reported the potential application of hyperspectral imaging in the visible and near infrared regions (4001090 nm) for nondestructive determination of MC in foods (strawberries and maize kernels [22, 23]). The development of a hyperspectral imaging (HSI) system as an analytical on-line tool to measure mushroom moisture content would enable rapid and nondestructive assessment of quality and management of product variability. The 4001000 nm wavelength range is industrially relevant due to the wide availability and affordable cost of charge-coupled-device (CCD) sensors in this range. Gowen et al. [24] investigated the quality attributes of sliced mushrooms including moisture content during storage using hyperspectral imaging. However this work was focused on a narrow range of MC values (9093%). The present research extends this work, by investigating the potential of hyperspectral imaging for the prediction of whole mushroom moisture content over a wider range of MC levels (6093%), which is more relevant to the typical MC variation observed in packaged whole mushrooms stored under typical industry conditions.

Ireland) at 45 1 C. Samples were removed from the oven at intervals of 0, 30, 60 and 120 min and stored for 30 min in a desiccator prior to weighing and hyperspectral image acquisition. The studied MC range was selected according to the typical MC variation for mushrooms packed in PolyPropylene (PP) trays and over-wrapped in Polyvinyl Chloride (PVC) (as is common packaging practice in the mushroom industry [25]) after a duration of one week at 19 C and relative humidity (RH) of 38% (MC at day 0 = 93.40 0.62%, MC at day 7 = 62.72 1.93% wb). Moisture content of each mushroom was measured using the oven method; samples were dried in a hot air oven at 110 C for 48 h [6] and moisture content (MC), evaluated by mass difference, was expressed as a wet base percentage (% wb). Colour measurement Previous research has reported that the Hunter L-value is an important mushroom quality parameter [2629]. Hunter L, a and b values of 10 mushrooms were measured, using a Minolta Chromameter (CR-400, Minolta Corp., Japan), at each experimental intervals (0, 30, 60 and 120 min) on the centre of the mushroom cap. Three readings were obtained and averaged for each mushroom. Hyperspectral imaging system The hyperspectral imaging system (Fig. 1) used in this research consisted of a high performance CCD camera (BASLER vision technologies, Germany) (580 9 580 pixels) covering the spectral range between 400 and 1000 nm, a spectrograph (Specim V10E, Finland) attached to the camera, a zoom lens, a light source transmitted

Materials and methods Sample preparation Agaricus Bisporus spp. mushrooms were grown in plastic bags and tunnels in Kinsealy Teagasc Research Centre (Malahide, Co. Dublin, Ireland). Spawn running and casing took place throughout the 6 weeks prior to mushroom cropping. Forty-eight blemish free second ush mushrooms, each with a diameter of 35 cm, were harvested in November (calibration set) and December 2008 (test set). Initial mass was noted and mushrooms were dried to four MC levels (93.40 0.62%, 82.76 2.11%, 73.20 2.60% and 60.89 4.32% wb) using a convective air dryer (Gallenkamp Plus II Oven, AGB Scientic, Dublin,

Fig. 1 The pushbroom hyperspectral imaging system used in the study (DV Optics, Italy)

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through ber optics, a moving table and a computer system equipped with Spectral Scanner software (DV Optics, Italy) to acquire the images. The noise characteristics of the sensor were investigated by acquiring 50 scans of the calibration tile over a time period of one hour. Signal to noise ratio was lowest at the upper (9501000 nm) and lower (400445 nm) wavelength limits; in these regions the noise level exceeded 1% of the signal. This is due to decreased CCD detector sensitivity in these regions. Due to this noise subsequent analysis of spectra was performed only on data in the 445 945 nm range. Average reectance (R) spectra from the total surface area were calculated for each sample using MATLAB 7.0 (The Math Works, Inc. USA). Reectance calibration In order to account for the non-linear sensitivity of the CCD camera a two point reectance calibration was performed (Ariana, Lu and Guyer, 2006). The bright response (W) was obtained by acquiring a hypercube from a uniform white ceramic tile (the reectance of which was precalibrated against a tile of certied reectance [Ceram Research (N.00608 of 28 May 2004)]; the dark response (D) was acquired by turning off the light source, completely covering the lens with its cap and recording the camera response. This was done prior to image acquisition at each time point. The corrected reectance value (R) was calculated from the measured signal (I) on a pixel-bypixel basis as shown below [30]: Ri Ii Di=Wi Di where i is the pixel index, i.e. i = 1, 2, 3,, n and n is the total number of pixels. Data processing and analysis In order to separate the mushroom from the image background, the hyperspectral images at each wavelength were pre-processed by masking [31]. The mask was created by thresholding the mushroom image at 800 nm (images at this wavelength provided good contrast between mushroom and background) and setting all background regions to zero. Non-zero elements of the image were then extracted and the mean spectrum was calculated for each mushroom image. Partial least square regression (PLSR) is a commonly used chemometric method for constructing models when the measured variables are many and highly collinear. In PLSR, the matrix of spectral values is transformed into principal components (which are linear combinations of the original spectral data) in order of decreasing variance; the principal component scores are used to estimate latent variables from the measured spectra to predict the variable

of interest (moisture content in this case). One benet of this method is that the PLS latent variables are uncorrelated and this typically leads to more stable predictions on new data. In this study PLSR was applied to the mean spectra using the pls package in R [32] to investigate the correlation between the spectral response and moisture content of mushroom samples and thereby predict mushroom moisture content. Leave-one-out (LOO) cross validation was applied to the calibration set and RMSECV and RMSEP were obtained by calculating the square root of the mean of the sum of squared differences between predicted and measured MC values of the calibration and test sets, respectively. Scattering effects are frequently encountered when obtaining diffuse reectance spectra of solid and semi-solid materials [33]. To reduce the inuence of scatter effects and other sources of variations (e.g. differences in mushroom sample height), spectral pre-processing methods were used. These methods were: Multiplicative Scatter Correction (MSC), Standard Normal Variate (SNV) [33], rst and second order SavitskyGolay derivative smoothing [34]. MSC (also known as Multiplicative Signal Correction) aims to reduce the effects of scattering in a set of spectra by performing linear regression on a reference or target spectrum; in this case the mean spectrum of the calibration set was used as the target spectrum [35].

Results and discussion Colour analysis Figures 2, 3 and 4 show the average L, a and b values for mushrooms at different MC levels, respectively. Gormley and OSullivan [36] reported the relationship between different quality levels in mushrooms and L-value; mushrooms with an L-value greater than 93 were classied as excellent,

Fig. 2 Average L-value of mushrooms at different moisture content levels (SD shown in error bars)

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of Variation, C.V. (Standard deviation/mean) was calculated for Hunter L, a and b values. It can be seen that the variation in a-value (C.V. = 116.37%) across the dehydration time is much greater than that of the b-value (C.V. = 20.01%) which in turn is greater than that of L-value (C.V. = 9.09%) suggesting the possible effectiveness of the a-value for monitoring quality attributes in mushroom. Hyperspectral image analysis Average reectance spectra obtained from the hyperspectral imaging data of the mushroom samples at different MC levels are shown in Fig. 5. It can be seen that average reectance decreases as moisture content decreases, however the spectral shapes appear to be similar at all MC levels. Standard deviation of the sample spectra at different MC is also plotted. The difference in reectance values between different treatments is relatively high across the wavelength range between 445 to 700 nm while from 700 to 945 nm the curves become similar and standard deviation curves are fairly similar in this wavelength region. PLSR was applied to the calibration set of pre-treated data using leave-one-out (LOO) cross validation to estimate the optimal model for prediction of the moisture content of mushrooms. The PLSR models were built on pre-processed spectra in the wavelength range between 445 to 945 nm (101 wavelength variables). To select the best pre-treatment, PLSR was applied to the pre-treated data and RMSECV, RMSEP and R2 of all pre-processed data were calculated for both calibration and test set (Table 1). Models based on SNV and MSC pretreated spectra were found to perform similarly when applied to the test set, with relatively low RMSECV and RMSEP and high R2 in comparison to the other investigated pre-treatments. SNV may be preferred to MSC as a pre-treatment because it does not require the use of a target spectrum.

Fig. 3 Average a-values of mushrooms at different moisture content levels (SD shown in error bars)

Fig. 4 Average b-values of mushrooms at different moisture content levels (SD shown in error bars)

an L-value between 90 and 93 as very good, an L-value between 86 and 89 as good, an L-value between 80 and 85 as reasonable and L-value between 69 and 79 as poor. In addition, mushrooms with an L-value less than 80 would not be acceptable at wholesale level, and those with an L-value less than 69 would not be acceptable at retail level. Mushrooms with an MC greater than 82.76% wb had an L-value greater than 80 indicating the acceptable quality of these samples. Mushrooms dehydrated to 73.20% and lower MC levels had a mean L-value less than 80 (mean L = 79.87 6.96 and 77.92 6.89), indicating potential unacceptability of these samples. Figure 3 shows that the avalue (redness) increased with drying time, indicating that a decrease in MC results in an increase in redness. The largest increase in a-value (from -0.162 to 4.535) was observed when the L-value approached the critical quality acceptance threshold (as dened by an L-value of 80). The b-values were positive for all MC levels and therefore refer to the yellowness of the mushrooms. The major changes in b-value occurred during the rst 60 min drying, with an increase of 63.7% observed, while in the nal 60 min drying, the b-value only increased slightly (by 0.31%). The Coefcient

Fig. 5 Average reectance spectra and the corresponding standard deviation for white button mushrooms at different moisture contents (% wb)

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Prediction of white button mushroom Table 1 R2 and RMSEP of different spectral treatments for the 4-component PLSR model Pretreatment Calibration set RMSEP (% wb) R Non pre-treated First derivative Second derivative SNV MSC 6.03 5.28 5.22 5.50 5.49
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Test set RMSEP (% wb) 9.88 5.82 8.51 5.58 5.57 R2 0.79 0.73 0.72 0.83 0.83

0.73 0.82 0.82 0.81 0.81

Figure 6 shows the RMSECV and RMSEP plotted as a function of the number of PLS latent variables for SNV pretreated data. For the calibration set, it can be observed that RMSECV decreases as the number of latent variables increases up to 13 whereas for the test set, it reaches a local minimum at four latent variables. This indicates that the correct number of latent variables should be no more than four. The rapid increase in RMSEP after 10 latent variables highlights the potential risk of over-tting the data when using a high number of latent variables. The minimum RMSECV and RMSEP values of 5.50 and 5.58% were obtained with a 4-component PLSR model built on SNV pre-treated reectance spectra. In order to compare these error values with previously reported data it is necessary to take the standard deviation of the sample sets into account. We recorded the ratio of percentage deviation (RPD) which is the ratio of the standard deviation to RMSECV or RMSEP [37]. RPD values between 1.8 and 2.0 indicates good model and/or predictions; RPD between 2.0 and 2.5 indicates very good model and/or predictions and RPD greater than 2.5 indicates excellent model and/or predictions [38]. The RPD values obtained in this study were 2.12 and 2 for the calibration and test sets, respectively. This compares favourably with the previously reported data on prediction of MC in mushrooms using

spectra in the 4001000 wavelength range. Roy et al. [6] reported SECV of 0.840.93 and standard deviation in MC of 2.89, giving an RPD of 3.13.44 for a 10 component PLS model. Gowen et al. [24] reported RMSEP of 0.74 and standard deviation in MC of 1.48, giving an RPD of 2 for mushroom slices. The predicted MC values for the developed 4-component PLSR model applied to the calibration and test sets of SNV pretreated data are plotted against actual MC in Figs. 7 and 8. There is a considerable and relatively uniform scatter of data points in the vertical direction among samples of similar moisture content for both calibration and test sets. This leads to a relatively low R2 for the model applied to the calibration and test sets (i.e. 0.81 and 0.83, see Table 1). This scattering indicates variability in the predicted response for samples exposed to the same drying

Fig. 7 Predicted MC as a function of actual MC for 4-component PLSR model built on the calibration set

Fig. 6 RMSECV and RMSEP as a function of the number of latent variables included in the PLSR model built on the SNV pretreated reectance spectra of the calibration set data (a) and applied to the test set data (b)

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Fig. 8 Predicted MC as a function of actual MC for 4-component PLSR model applied to the test set

conditions. The predicted variable in this case is bulk moisture content; however, the spectra obtained from the hyperspectral imaging system characterise mainly the surface properties of the mushroom. The observed vertical shifts in the predicted response may therefore be related to differences in the surface moisture content of samples, which may vary from sample to sample even though they have the same bulk moisture content. In order to demonstrate model performance over the surface of the mushroom, MC prediction maps of
Fig. 9 Prediction maps for PLSR predictive model applied to mushroom hyperspectral data for a fresh mushroom, b 30 min dried mushrooms, c 60 min dried mushrooms, d 120 min dried mushrooms

mushrooms dried for different time periods were constructed by applying the 4-component PLSR model to SNV pre-treated hyperspectral images. To reduce processing time, hypercubes were reduced to 10% of their original size using bi-cubic interpolation. In order to apply the procedure, it was necessary to rst unfold the hypercube into a two-dimensional matrix in which each row represented the spectrum of one pixel. The SNV transformation was applied to the unfolded spectra, after which the 4-component PLSR model (built on the calibration set) was applied. The resultant matrix was refolded to form a prediction image as shown in Fig. 9. The average pixel value of each predicted image, which represents the predicted moisture content for each mushroom image, was calculated and is also shown. Overall, the prediction maps for mushrooms with different MC levels show that the model preformed well for prediction of mean mushroom moisture content in the range studied. Using HSI in this way differentiates areas of different moisture content enabling better understanding of dehydration distribution over the mushroom surface. However, the curvature of the mushroom surface may contribute to the dark regions at the mushroom edges predicted as having lower MC than the central regions.

Conclusions The optimal pre-treatment for predicting mushroom moisture content was the SNV transformation. PLSR models were developed to predict MC of mushrooms with a

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4-component PLSR model giving R2 value = 0.81 and RMSECV = 5.50 for the calibration set and R2 value = 0.83 and RMSEP = 5.58 for the test set. The regression model performed well for prediction of MC in the range relevant to the mushroom industry (6093%). MC prediction maps were generated to estimate the distribution of water over the mushroom surface. This study demonstrates the potential use of hyperspectral imaging as an analytical on-line tool to measure mushroom moisture content. Further development of such a tool would facilitate rapid assessment of mushroom shelf life and management of product variability.
Acknowledgements The authors would like to thank Dr. Helen Grogan and Ted Cormican from the Teagasc Research Station at Kinsealy, Dublin, for production of mushrooms and advice. This research was funded by the Irish Government Department of Agriculture and Food under the Food Institutional Research Measure (FIRM).

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