Bioresource Technology 102 (2011) 40834090

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Evaluation of near infrared spectroscopy and software sensor methods for determination of total alkalinity in anaerobic digesters
Alastair J. Ward a,, Philip J. Hobbs b, Peter J. Holliman c, David L. Jones d
a

Department of Biosystems Engineering, Faculty of Agricultural Science, University of Aarhus, Blichers All 20, 8830 Tjele, Denmark North Wyke Research, North Wyke Research Station, Devon EX20 2SB, UK c School of Chemistry, Bangor University, Bangor, Gwynedd LL57 2UW, UK d School of Environment, Natural Resources and Geography, Bangor University, Bangor, Gwynedd LL57 2UW, UK
b

a r t i c l e

i n f o

a b s t r a c t
In this study two approaches to predict the total alkalinity (expressed as mg L1 HCO 3 ) of an anaerobic digester are examined: rstly, software sensors based on multiple linear regression algorithms using data from pH, redox potential and electrical conductivity and secondly, near infrared reectance spectroscopy (NIRS). Of the software sensors, the model using data from all three probes but a smaller dataset using 2 total alkalinity values below 6000 mg L1 HCO 3 produced the best calibration model (R = 0.76 and root ). When validated with new data, the NIRS mean square error of prediction (RMSEP) of 969 mg L1 HCO 3 method produced the best model (R2 = 0.87 RMSEP = 1230 mg L1 HCO 3 ). The NIRS sensor correlated better with new data (R2 = 0.54). In conclusion, this study has developed new and improved algorithms for monitoring total alkalinity within anaerobic digestion systems which will facilitate real-time optimisation of methane production. 2010 Elsevier Ltd. All rights reserved.

Article history: Received 10 September 2010 Received in revised form 8 December 2010 Accepted 9 December 2010 Available online 22 December 2010 Keywords: Biogas Anaerobic digestion Monitoring NIRS Software sensor

1. Introduction The anaerobic digestion process can be unstable, particularly when subjected to changes in the fermentation environment, for example following an increase in inuent concentration (Moreira et al., 2008; Ward et al., 2008), a reduction in hydraulic retention time (HRT) (Converti et al., 2008) or a change in the nature of the feedstock (Nain and Jawed, 2006). Such variations often have the effect of reducing methane production in the digester. It is therefore common in industrial applications to construct a digester that is larger than the optimal size to reduce the impact of process instability (Jantsch and Mattiasson, 2004). However, this option is expensive in terms of construction, operation and maintenance costs. A preferable alternative would be to continuously monitor the key variables within the process and to use this information to make decisions regarding the loading of the plant, either through automatic organic loading rate control systems or advice to operators. This also makes sound economic sense in terms of reduced capital costs and improved gas output. There have been several publications which suggest alkalinity as an ideal parameter to monitor the anaerobic digestion process; as total alkalinity (Fernandez et al., 2008), partial alkalinity (Kyazze et al., 2006), the ratio of partial to total alkalinity (Mendez-Acosta
Corresponding author. Tel.: +45 8999 1935; fax: +45 8999 1619.
E-mail address: alastair.ward@agrsci.dk (A.J. Ward). 0960-8524/$ - see front matter 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.biortech.2010.12.046

et al., 2010) or the ratio of volatile fatty acids (VFA) to alkalinity (Barampouti et al., 2005). Total alkalinity includes the VFA buffering system whereas partial alkalinity predominantly measures the bicarbonate concentration. It has been suggested that partial alkalinity is more sensitive than total alkalinity for detecting process imbalances (Jantsch and Mattiasson, 2003) but it has been shown that maintaining total alkalinity is sufcient to prevent a reduction in reactor pH (Borja et al., 2004; Fernandez et al., 2008). Alkalinity is normally measured ex situ by titration with a pH probe (Ferrer et al., 2010; Nges and Liu, 2010), but other methods include: a spectrophotometric method using coloured pH indicators (Jantsch and Mattiasson, 2003, 2004), mid infrared spectroscopy (Steyer et al., 2002) or acidication of a sample and subsequent measurement of the volume of carbon dioxide produced (Hawkes et al., 1993). However, these analytical procedures either contain several individual steps, time consuming sample preparation or the use of various reagents to determine alkalinity or process stability. Chemical reagents for on-line instrumentation can also add to plant operating costs. For instance, the estimated reagent costs for an on-line total organic carbon analyser for use in biogas processes was approximately 4000 per year in 2002 (Steyer et al., 2002). The anaerobic digestion process can also be monitored by less direct methods which rely on mathematical models to determine key parameters. Such methods include software sensors and near infrared spectroscopy (NIRS).

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Software sensors use mathematical models which utilise easily measured parameters to estimate important parameters which are difcult or even impossible to measure directly. Alcaraz-Gonzalez et al. (2002) used a wide variety of inputs including input ow rate, carbon dioxide exhaust ow rate, fatty acid concentration and total inorganic carbon to estimate the unknown parameters of microbial concentrations, alkalinity and chemical oxygen demand in a waste water treatment plant. Furthermore, Feitkenhauer and Meyer (2004) estimated substrate and bacterial biomass concentrations from inputs based on titrimetric techniques, and Bernard et al. (2000) used a mass balance based model and gaseous measurements to predict fatty acids and inorganic carbon, and a separate software sensor to estimate bacterial biomass. Near infrared spectroscopy relies on multivariate techniques such as partial least squares regression models to determine multiple parameters simultaneously. NIRS has shown some success when monitoring anaerobic digestion processes (Hansson et al., 2002, 2003; Holm-Nielsen et al., 2008; Jacobi et al., 2009), particularly for the determination of volatile fatty acids (VFA). Advantageously, NIRS methods can measure several parameters simultaneously if a calibration model has been made for the parameter(s) of interest. An NIR spectrometer is in contact with the digestate via a window or probe, often constructed from sapphire or a similar scratch resistant material and hence is low maintenance. Similarly, electrode-type probes such as pH and redox potential contain no mechanical parts and only require occasional calibration and cleaning (the latter being built into the design of many industrial-scale instruments). Such approaches may provide more robust and reliable monitoring systems without the need for reagents or sample preparation. In summary, no simple probe exists to measure total alkalinity directly within an anaerobic digester, yet a combination of hardware sensors can be used to indirectly predict total alkalinity by developing an algorithm from regression analysis on the sensor data. This paper examines two analytical approaches using multivariate techniques for the determination of total alkalinity in an anaerobic digester in real time. First, a software sensor which derives total alkalinity from pH, redox and conductivity data and second a NIR spectrometer that derives a calibration curve on the principal components of the spectra. A single substrate was used throughout the fermentation monitoring period and a reduction in hydraulic retention time was used as the source of disturbance to obtain a wider range of total alkalinity values.

Carbon to nitrogen ratio was determined using a Carlo Erba NA2000 Elemental Analyser (Carlo Erba Reagenti SpA, Rodano, Milan). Crude protein was calculated by multiplying the nitrogen percentage by 6.25 (Gizachew and Smit, 2005). Total and volatile solids were determined by standard methods (APHA, 1976). 2.2. Digester construction A digester of four sequential stages with a total volume 220 L was constructed for data collection to construct multiple linear regression and partial least squares NIRS models. The vessels will henceforth be referred to as V1, V2, V3 and V4, in respective order of their position in the chain. Feeding of the research digester was automatic and took place every hour via a peristaltic pump (Watson-Marlow 323 D/u, Watson-Marlow Bredel Pumps, Falmouth, Cornwall, UK) controlled by a time switch. Feeding of the rst vessel caused an increase in the liquid level that, in turn, led partially digested feedstock to enter the subsequent vessel (V2, V3 or V4) by a weir system; the overall level of the system being governed by the height of the weir outlet of V4. The fourstage digester was split into two 2-stage digesters; one of which was used for testing of the software sensor and NIRS models. The four stage digester was designed to produce a large quantity of data and to observe digestion as a progression through the four vessels. The system was operated under a variety of loading conditions, including start-up, failure, recovery and steady state operation over a period of approximately 9 months. Measurement of pH, oxidationreduction potential, electrical conductivity (EC) and temperature was performed by Partech Waterwatch 2610 ow cells (Partech Instruments, St Austell, Cornwall, UK). The probes had measurement ranges of 014 units for pH, 700 to +700 millivolts (mV) for redox potential and 010 millisiemens per centimetre (mS cm1) for EC. Three of these ow cells and their corresponding control/display boxes were used, placed between V1 and V2, V2 and V3 and V3 and V4. These units produced 420 mA outputs which were connected to a National Instruments SCB68 data acquisition interface (National Instruments Corporation Ltd., Newbury, Berkshire, UK) which, in turn, supplied digital control signals to a National Instruments PCI-6229M Series Multifunction DAQ Device, installed in a PC computer. LabVIEW v.7.1 software with NIDAQ Measurement Services Software (for data acquisition), also from National Instruments, were used to record probe data. Samples for titration and NIR analysis were taken from a bulk sample of approximately 100 mL collected from the ball valves located on each of the ow cells. The bulk sample was well mixed before splitting into sub samples for NIR analysis as well as a further sub sample for titration purposes. The samples were in this way all from the same bulk sample and therefore each sub sample was highly representative of the bulk sample. In addition, the bulk sample was collected from a point directly below the probes and was therefore material which was directly in contact with the probes immediately before sampling. Gas production of each vessel was measured by water displacement, with equipment built in-house. Methane content of the biogas was measured by infrared spectroscopy using a hand held analyser (Crowcon Triple + plus IR, Crowcon Detection Instruments Ltd, Abingdon, Oxfordshire, UK). 2.3. Determination of total alkalinity (HCO 3 concentration) Total alkalinity analysis was conducted using an auto-titrator (Metrohm 716 DMS Titrino, Metrohm House, Buckingham, UK) with a two point alkalinity titration from pH 4.54.2 using 0.02 M H2SO4; the titrator software automatically calculating slope and equivalent CaCO3 concentration. This was multiplied by 1.22

2. Methods 2.1. Inoculum and feedstock composition The digester was inoculated with digested material from a large scale centralised biogas plant (AnDigestion Ltd. Chilsworthy, Holsworthy, Devon, UK) treating a mixture of cattle manure and food industry waste. This inoculum was characterised for total solids, volatile solids and total alkalinity. Commercially available pig feed was used as a feedstock as this was easily available and of consistent composition. The feed was ground to 62 mm and mixed with tap water to prepare a feedstock of 5% (w/v) total solids. The feedstock was primarily composed of cereals and was analysed for neutral detergent bre (NDF), acid detergent bre (ADF) and acid detergent lignin (ADL) using an Ankom 2000 bre analyser (Ankom Technology, Macedon, NY, USA) and methods based on Van Soest et al. (1991). After analysis, the samples were incinerated to determine ash content. These values were used to determine hemicellulose (NDFADF), cellulose (ADFADL) and lignin (ADLash) content.

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to achieve the HCO 3 equivalent concentration. All titrations were performed in duplicate. 2.4. Non linear regression models for methane production The titrated total alkalinity data was non-linearly regressed against the measured methane production rates to nd the relationship between total alkalinity and methane production rate and subsequently to nd the total alkalinity level at which methane production was maximised. A variety of curve shapes were tted to the data, quadratic regression curves were found to be the best t. Optimal total alkalinity was determined for V1, V2, V3 and V4 separately due to differences in the magnitude of methane production from the different vessels. The curve tting models were created using Genstat v.9.2 software (Lawes Agricultural Trust, Rothamsted Experimental Station, Harpenden, UK).

2.5. Principle component analysis of probe data The dataset used for the software sensor models was initially subjected to principal component analysis (PCA). This technique examines variation in the matrix of the probe data (the X matrix) and decomposes the X matrix into structural and noise components. The greatest variation in the data matrix is considered to be the rst principal component and the second greatest variation, principal component 2, is plotted orthogonally to component 1 and so on for further principal components. Higher order principal components progressively describe more noise and less structure in the X matrix. Due to the differences in order of magnitude between redox potential and pH/conductivity, the data were normalised before PCA by multiplying all values by 1/standard deviation. The models were validated using leave one out cross validation, where a single sample is left out of the calibration and used to validate the model. This process is then repeated with a different sample used for validation and so on until all samples have been used for validation. 2.6. Regression models for the prediction of total alkalinity from probe data Data from the pH, redox and conductivity probes at the time of sample collection were tted to the titration total alkalinity data through linear regression and multiple linear regression (MLR) models of single probe data and combined probe data (respectively) using The Unscrambler v.9.8 software (CAMO Software A/ S, Oslo, Norway). All models were validated using leave-one-out cross validation as described in Section 2.5. 2.7. Near infrared spectroscopy All samples were analysed with a Bruker Vector 22/N FT-NIR spectrometer with OPUS v. 5.0 software for spectral analysis and OPUS Lab software as a user interface for measurement of multiple samples (Bruker Optics Limited, Coventry, UK). In a typical analysis, 20 mL of each fresh sample was measured in triplicate at room temperature in 40 mL glass vials. Each sample was measured in triplicate and the spectra were later averaged. The frequency range of the spectrometer was 12,8004000 cm1 (k = 7802500 nm) with the resolution set at 16 cm1. The OPUS quantitative analysis software uses a partial least squares (PLS) regression which is commonly used in NIR spectral analysis (McCarty et al., 2002; Viscarra Rossel et al., 2006). The large number of samples (>230 in triplicate) enabled the use of test set validation of the model, whereby two thirds of the samples were used to build the model and the remaining samples were used to validate the model. Every third sample in a chronologically ordered list was used for validation. The software included an automatic optimisation option which constructed various PLS models from different regions of spectra and with different spectral

(a) 1.8
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1.4 1.2 1 0.8 0.6 0.4 0.2 0 0 3000 6000

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Fig. 1. (a) Non-linear regression quadratic curve of total alkalinity against methane production rate for the rst anaerobic digester vessel (V1). Other vessels showed a similar optimal alkalinity level for maximising methane production. (b) Principal component analysis score plot based on the full probe dataset, the symbols represent total alkalinity grouped as follows: Level 1, 03000 mg L1 HCO 3 , Level 2, 1 >30006000 mg L1 HCO HCO 3 , Level 3, >6000 mg L 3.

Table 1 Model statistical results and MLR algorithms for models using combinations of pH (P), redox (R) and electrical conductivity (C) data, ordered in terms of best model at the top for full range datasets, where Alkp = predicted total alkalinity in mg L1 HCO 3, Vp = pH value (pH units), Vr = redox value (mV) and Vc = conductivity value (mS cm1). Sensor combination PC PRC P PR RC C R R2 0.68 0.68 0.58 0.58 0.44 0.41 0.13 RMSEP (mg L1) 1419 1421 1608 1614 1868 1936 2329 Equation Alkp Alkp Alkp Alkp Alkp Alkp Alkp 7997 1465 V p 374:219 V c 7965 1483 V p 0:459 V r 373:46 V c 7794 1868 V p 7801 1872 V p 0:061 V r 1407 3:733 V r 615:93 V c 689 687 V c 2346 6:703 V r

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pre-treatments. The best model was recommended based on minimising the root mean square error of prediction (RMSEP, the average error of the prediction performed during validation) and maximising the R2 (variance accounted for by the model) value. 3. Results and discussion 3.1. Inoculum and feedstock composition The inoculum was screened for particles >2 mm which would have caused blockage problems in the digester. The screened inoculum was measured as 5.6% total solids, 92.4% volatile solids and had a total alkalinity of 24,500 mg L1 HCO 3. The dry feedstock had a total solids value of 94.3% (diluted with tap water to 5% total solids for use) and a volatile solids value of 93.9%. Total carbon was 40.7% and total nitrogen was 2.3%, giving a C:N ratio of 17.6. From the bre analysis, hemicellulose was determined as 28.3%, cellulose 21.3% and lignin as 8.2% of the dry material. 3.2. Optimisation of methane production rate The non-linear regression curve of total alkalinity against methane production rate for V1 is shown in Fig. 1a. Optimal methane production was found at alkalinities between approximately 3000 and 6000 mg L1 HCO 3 . V2, V3 and V4 were found to have similar optima. From the curves, it was clear that methane production fell sharply below a total alkalinity level of ca. 3000 mg L1 HCO 3 , indicating failure of the process below this concentration. Methane production also dropped when total alkalinity exceeded ca. 6000 mg L1 HCO 3 , suggesting that total alkalinity is an important factor affecting methane production. 3.3. Principal component analysis The resulting PCA model can be visualized as a two dimensional score plot which plots objects in relation to two principal components to show groupings and/or relationships (Fig. 1b). The score plot presents samples in relation to the rst two principal components as these represent 63% and 24% (respectively) of the total variation in the X matrix. The titrated total alkalinity data was not included in the analysis but these values were converted into category variables with the purpose of grouping the samples in the score plot. The categories were grouped logically according to the ndings discussed in Section 3.2: Level 1 was 03000 mg L1 HCO 3 , which corresponded to an overloaded and failing digester, Level 2 was >30006000 mg L1 HCO 3 which indicated methane production was optimal and Level 3 was >6000 mg L1 HCO 3 , indicating the digester was under loaded. It is clear from Fig. 1b that Level 1 samples are positioned to the left of the score plot. This means that the largest variation in probe data is also related to total alkalinity and is accounted for by the rst principal component. Level 2 and 3 samples are less clearly grouped in the score plot. The PCA demonstrates that the data collected by the probes may be better capable of distinguishing between Levels 1 and 2/3 but less suited to distinguishing between Levels 2 and 3. This was acceptable, as it was considered more important to detect an overloaded and failing digester than an under loaded digester. 3.4. Software sensor algorithm model development Table 1 shows the software sensors regression model calibration data including R2, RMSEP and the regression model equations, in descending order of model quality from the top downwards. RMSEP is expressed as total alkalinity in mg L1 HCO 3 . The models

were named based on the probe(s) used: P for pH, R for redox potential and C for conductivity. Fig. 2a shows a predicted versus measured plot of the best calibration model, the two probe PC model, in terms of R2 and RMSEP. The model statistics and equation (Table 1) are very similar to those of the three probe PRC model suggesting that redox potential played little part when

Fig. 2. (a) Regression model PC using full dataset. (b) Regression model PRC<6000 using a calibration dataset with total alkalinity below 6000 mg L1 HCO 3 . (c) 10 Principal component NIRS regression model. (d) 13 Principal component NIRS regression model.

A.J. Ward et al. / Bioresource Technology 102 (2011) 40834090 Table 2 Model statistical results and MLR algorithms for models using combinations of pH (P), redox (R) and electrical conductivity (C) data ordered in terms of best model at the top for datasets restricted to total alkalinity values of less than 6000 mg L1 HCO 3 , where Alkp = predicted 1 total alkalinity in mg L1 HCO ). 3 , Vp = pH value (pH units), Vr = redox value (mV) and Vc = conductivity value (mS cm Sensor combination PRC<6000 PC<6000 PR<6000 P<6000 RC<6000 C<6000 R<6000 R2 0.76 0.75 0.63 0.62 0.59 0.46 0.29 RMSEP (mg L1) 969 989 1207 1229 1280 1470 1678 Equation Alkp Alkp Alkp Alkp Alkp Alkp Alkp 5171 893 V p 2:854 V r 289:97 V c 5640 1099 V p 302:123 V c 4673 1118 V p 3:331 V r 5200 1371 V p 1277 6:283 V r 385:188 V c 30:44 489:046 V c 862 8:223 V r

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determining total alkalinity. This was reected in the results for the regression model predicting total alkalinity from redox potential alone, which had the lowest R2 and highest RMSEP of all the models shown in Table 1. The models in Table 1 were found to have a poor response for total alkalinity values above approximately 6000 mg L1 HCO 3. This poor ability to model high total alkalinity values was also evident in the PCA results and was attributed to the fact that as total alkalinity increased above ca. 6000 mg L1 HCO 3 , pH (where applicable) did not rise signicantly and that the conductivity probe (where applicable) often exceeded the maximum range of 10 mS cm1. To improve on the models in Table 1, new models were created based on measured (titrated) total alkalinity values of less than 6000 mg L1 HCO 3 only. The revised regression models are shown in Table 2, again ordered in descending order of model quality from the top downwards. The smaller dataset models shown in Table 2 in all cases improved upon the model parameters considerably, for example RMSEP in most cases is ca. 35% less with the smaller datasets. When using the smaller dataset the three probe PRC<6000 model (Fig. 2b), performed slightly better than the PC<6000 model but as was the case for the full dataset models (Table 1) the difference was small which further claried the limited effect of redox potential on the models. 3.5. NIRS model development The NIRS model was derived from a larger sample set than the software sensor model, with a greater range of total alkalinity (maximum total alkalinity of ca. 22,400 mg L1 HCO 3 for the NIRS model compared to ca. 11,600 mg L1 HCO for the software sensor 3 model). This was due to the software sensor probe data acquisition systems not being completed during the four-stage digester early start-up period. The start-up period utilised an inoculum with a high total alkalinity which greatly extended the range of measured values. Model optimisation used a maximum rank of 15 (i.e. the model was restricted to a maximum of 15 principal components). The OPUS Quant optimisation software calculated and arranged all suitable PLS models in order of model quality, the second best model (Fig. 2c) used 10 principal components and two specic bands of spectra found between 9751.27498.5 and 6102.2 4246.8 cm1. The software also suggested that spectral pre-processing was required in the form of straight line subtraction. This process subtracts a straight line from the spectrum, thus removing any tilt in the absorbance values across the range of wavenumbers due to differing energy absorbance over such a large energy range. The resulting model had an R2 of 0.84 and a RMSEP of 1390 mg L1 HCO 3 after test set validation and 11 outliers were found by the software. The residual prediction deviation (RPD) was calculated by dividing the standard deviation of the calibration dataset by the RMSEP. The RPD allows comparison of the RMSEP of models

of very different values to be compared. The calculated RPD was 2.92, just below the recommended minimum of three suggested as acceptable for quantitative determination by Williams (2001). Fig. 2d shows the prediction versus measured plot of the 13 principal component model which had highest R2 and lowest RMSEP, although the high number of principal components made the model more complex. The NIRS models were made with an offset of zero and a slope of one. The 13 principal components model had an R2 of 0.87 and a RMSEP of 1230 mg L1 HCO 3 with only one outlier highlighted by the software. The optimisation procedure that produced this model did so by only considering wavenumbers between 7502.4 and 4246.8 cm1 and vector normalisation as spectral pre-processing. Vector normalisation calculates the average intensity value of the chosen spectra and subtracts this value from the entire section(s) of the spectrum. The sum of the squared intensities is then calculated and the spectrum is divided by the square root of this sum (Bruker, 2004). The RPD for the thirteen principal components model was 3.3, which was above the minimum recommended criteria (Williams, 2001) therefore the model was considered valid. The detection of outliers requires that the spectral matrix X be inverted and bi-diagonalised. This allowed calculation of the leverage value (hi) as shown in Eq. (1) (Bruker, 2004).

hi diagUU T

where U represents orthonormal matrices. The leverage was a measure of how much inuence a particular spectrum had on the PLS model for a particular component. Leverage values are always less than 1 and the sum of all leverages was equal to the rank (R) as shown in Eq. (2) (Bruker, 2004).

hi R

From this, the mean leverage value, R/M was calculated, where M was the number of calibration samples. It was recommended that 5 R/M was a suitable limit for determining outliers (Bruker, 2004). No other NIRS predictions of total alkalinity have been published to the authors knowledge and therefore the model parameters are difcult to compare. However, they compare favourably with recent NIRS predictions of VFA in biogas processes (HolmNielsen et al., 2008; Jacobi et al., 2009). 3.6. Validation of software sensor and NIRS models The two stage digester used for validation of the total alkalinity models provided 44 samples over 60 days for titration and NIRS analysis, in addition to the probe data collected in-line in real time for validating the software sensor models. The digester was overloaded by decreasing the HRT towards the end of the experiment to observe the behaviour of the sensors as total alkalinity fell. The pH, redox and conductivity data recorded at the time of sam-

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Total alkalinity (mg L -1 HCO 3-)

pling were used to calculate the software sensor total alkalinities for all models using the equations in Tables 1 and 2. The NIRS and software sensor total alkalinity estimates were plotted against the titrated alkalinities and the slopes, offsets and R2 values are shown in Table 3, arranged in descending order of model quality from top to bottom. Standard deviations between triplicates of the NIRS predicted values were less than 10% of the mean. The R2 values for the validation data were lower than for the original model data. The best performing model was the NIRS 13 principal components model in terms or R2 although the NIRS 10 principal components model had a slope closer to one and a lower offset value. Also of note in Table 3 is the performance of the software sensor models which used the full and smaller (total alkalinity < 6000 mg L1 HCO 3 ) datasets. The larger dataset models were found to be more accurate at predicting new values than the smaller dataset models, despite the lower quality of the formers original calibration models. Therefore there was an advantage in using a calibration dataset which covered a greater range of values. It is also of importance that only 15 of the 44 new samples had a titrated total alkalinity of greater than 6000 mg L1 HCO 3 , thus the smaller dataset models were attempting to estimate values above those included in the calibration set. In addition, random sampling error or variance is often reduced with a larger number of samples (Esbensen et al., 2007). The three low NIRS predicted total alkalinity values at days 22, 23 and 24 (Fig. 3a) coincided with a pipe blockage problem within the system. The samples taken on these days produced NIR spectra of limited absorbance range when compared to the spectra of other samples and such unusual spectra would, in practice, be treated with suspicion. Removing these three data points increased the R2 values for both NIRS models and it is the modied values which are shown in Table 3. The low slope values for the small dataset models were a cause for concern as this suggests reduced sensitivity. Moreover, the high offset values of all models was less than ideal for estimating total alkalinity values of less than 3000 mg L1 HCO 3 (the total alkalinity level at which digester failure was expected to occur). The offset value in these data is the measure of the estimated total alkalinity at zero titrated (reference) total alkalinity. Thus, the off-set is effectively the limit of detection for each model. For many of the models in Table 3, the offset (and therefore the limit of detection) is greater than 3000 mg L1 HCO 3 . This explains why the NIRS rank 13 model (offset = 2736 mg L1 HCO 3 ) model best predicts the drop in total alkalinity below 3000 mg L1 HCO 3 because the linear range of this model extends into this concentration range. To better observe the response of the sensors to sudden changes in total alkalinity, the data estimated by the NIRS rank 13 model,
Table 3 Prediction model data for models using NIR and combinations of pH (P), redox (R) and electrical conductivity (C) data with both full range datasets and datasets restricted to total alkalinity values of less than 6000 mg L1 HCO 3. Sensor NIRS rank 13 PC PRC NIRS rank 10 PR<6000 P<6000 PRC<6000 PR P R<6000 PC<6000 R RC<6000 RC C C<6000 Slope 0.460 0.390 0.390 0.550 0.176 0.189 0.289 0.257 0.257 0.056 0.304 0.050 0.237 0.337 0.348 0.247 Offset 2736 3061 2984 2059 4355 3899 3171 4589 4599 4835 2734 5585 3135 2596 2293 1776.7 R2 0.54 0.40 0.39 0.36 0.30 0.28 0.24 0.21 0.20 0.08 0.08

the NIRS rank 10 model, the PRC<6000 model and the PC model were plotted against time and directly compared to the titrated total alkalinity (Fig. 3ad). The NIRS rank 13 model (Fig. 3a) followed the titrated reference data best, especially with regard to the decrease in total alkalinity below the critical 3000 mg L1 HCO 3 level

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Fig. 3. Prediction of total alkalinity over a 60 days validation period comparing the reference data (titration) with: (a) NIRS rank 10 model, (b) NIRS rank 13 model (c) PRC<6000 model, (d) PC model.

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from day 54 onwards. The NIRS rank 10 model (Fig. 3b) performed comparatively poorly during this period. The poor performance of the NIRS rank 10 model was attributed to under tting of the model due to too few principal components, i.e. too little of the spectral information was used in the model. Comparing the best of the small and full dataset calibration models (PRC<6000 and PC models, Fig. 3c and d respectively) shows that the full dataset PC model gives a better estimate of total alkalinity at the higher levels (above 6000 mg L1 HCO 3 ) and also at the critical lower level of below 3000 mg L1 HCO 3 . However, none of the software sensor models performed as well as the NIRS 13 principal component model. The software sensor was developed as an in-line system from the start and scaling up to industrial scale would be a very simple procedure at relatively low cost, requiring only the addition of the pH and conductivity probes into an existing anaerobic digester efuent outlet. The NIRS system described here is an off-line method but could easily be implemented as an in-line method with the use of a bre-optic probe positioned in a similar way as described above for the software sensor. However, an NIR spectrometer is considerably more expensive than the software sensor probes, at ca. 50,000 for the former compared to ca. 4000 for the latter. The strongest point of the two methods of total alkalinity determination described here is their low maintenance and lack of sample preparation rather than their absolute precision and accuracy. Thus, the aim is to track trends in total alkalinity to monitor the process and maintain total alkalinity between 3000 and 5000 mg L1 to optimise methane yields. This is actually a broad concentration range so the main requirement of the model is to be able to track changes in total alkalinity fast and accurately enough to enable rapid corrective action to be taken before methane yield drops. The NIRS method uses an internal reference system which requires no operator input and the software sensor probes only required cleaning and calibration every 2 months. Spanjers et al. (2006) found that a sample pre-treatment unit tted upstream of a mid-infrared spectrometer was subject to clogging on several occasions over a 6 months assessment period, yet both methods described here do not require sample pre-treatment. However, the methods described in this paper are not as accurate as other on-line methods described in the literature: Jantsch and Mattiasson (2004) achieved a model response with R2 = 0.99 for spiked, ltered efuent samples from a municipal sludge digester and Hawkes et al. (1993) measured bicarbonate to an accuracy of 5% in simulated ice cream waste water using the addition of sodium bicarbonate or dilution with water to achieve high and low alkalinities (respectively). It should be noted that both the software sensor models and the NIRS model described here could be specic to the feedstock used. It is expected that many intermediates in the anaerobic digestion process may be similar (for example VFA) but many other components could change the matrix and without validation on a different feedstock the possibility of universal applicability of these models cannot be guaranteed. The NIRS model could be utilising a large range of feedstock specic parameters for total alkalinity determination and this may also be the case for the conductivity probe, as the exact mechanism by which total alkalinity affects conductivity has not been fully established. Although it is understood that as pH rises above the pka of potentially ionisable species these will become ionised leading to an increase in conductivity. However, the concentration of these potentially ionisable species is likely to be different in any two digesters fed with different feedstocks, as the measurement of electrical conductivity in the water industry is known to be impossible to compare between different sample sources and, as such, is used only for comparative measurements from the same water source (HMSO, 1978).

4. Conclusions The software sensor models gave highest R2 = 0.76 (PRC<6000 model). Validation on new samples gave highest R2 = 0.40 (PC model). The NIRS model gave improved performance with R2 = 0.87. Validation of the NIRS model on new samples gave R2 = 0.54. Both methods allowed continuous monitoring but were limited in accuracy and precision which means that, without further development, they are only suitable for approximate indications of total alkalinity. However, because the optimum total alkalinity range for maximum methane production was found to lie between 3000 and 6000 mg L1 HCO 3 , the NIRS sensor shows promise as an on-line sensor to monitor anaerobic digestion processes.

Acknowledgements The authors would like to thank the European Social Fund and the UK Biotechnology and Biological Sciences Research Council for providing funds for this research.

References
Alcaraz-Gonzalez, V., Harmand, J., Rapaport, A., Steyer, J.P., Gonzalez-Alvarez, V., Pelayo-Ortiz, C., 2002. Software sensors for highly uncertain waste water treatment plants: a new approach based on interval observers. Water Research 36, 25152524. APHA, 1976. Standard Methods for the Examination of Water and Wastewater, 14 ed. APHA, Washington DC. Barampouti, E.M.P., Mai, S.T., Vlyssides, A.G., 2005. Dynamic modelling of the ratio of volatile fatty acid/bicarbonate alkalinity in a UASB reactor for potato processing wastewater treatment. Environmental Monitoring and Assessment 110, 121128. Bernard, O., Hadj-Sadok, Z., Dochain, D., 2000. Software sensors to monitor the dynamics of microbial communities: application to anaerobic digestion. Acta Biotheoretica 48, 197205. Borja, R., Rincon, B., Raposo, F., Dominguez, J.R., Millan, F., Martin, A., 2004. Mesophilic anaerobic digestion in a uidised-bed reactor of wastewater from the production of protein isolates from chickpea our. Process Biochemistry 39, 19131921. Bruker Spectroscopic Software OPUS Quant, 2004. Bruker Optics Limited, Banner Lane, Coventry CV4 9GH, England. Converti, A., Oliveira, R.P.S., Torres, B.R., Lodi, A., Zilli, M., 2008. Biogas production by means of a two-step biological process. Bioresource Technology 100 (23), 5771 5776. Esbensen, K.H., Friis-Petersen, H.H., Petersen, L., Holm-Nielsen, J.B., Mortensen, P.P., 2007. Representative process sampling in practice: variographic analysis and estimation of total sampling errors (TSE). Chemometrics and Intelligent Laboratory Systems 88, 4159. Feitkenhauer, H., Meyer, U., 2004. Software sensors based on titrimetric techniques for the monitoring and control of aerobic and anaerobic bioreactors. Biochemical Engineering Journal 17, 147151. Fernandez, N., Montalvo, S., Borja, R., Guerrero, L., Sanchez, E., Cortes, I., Colmenarejo, M.F., Travieso, L., Raposo, F., 2008. Performance evaluation of an anaerobic uidized bed reactor with natural zeolite as support material when treating high-strength distillery wastewater. Renewable Energy 33, 24582466. Ferrer, I., Vazquez, F., Font, F., 2010. Long term operation of a thermophilic anaerobic reactor: process stability and efciency at decreasing sludge retention time. Bioresource Technology 101 (9), 29722980. Gizachew, L., Smit, G.N., 2005. Crude protein and mineral composition of major crop residues and supplemental feeds produced on Vertisols of the Ethiopian highland. Animal Feed Science and Technology 119, 143153. Hansson, M., Nordberg, A., Sundh, I., Mathisen, B., 2002. Early warning of disturbances in a laboratory-scale MSW biogas process. Water Science and Technology 45 (10), 255260. Hansson, M., Nordberg, A., Mathisen, B., 2003. On-line NIR monitoring during anaerobic treatment of municipal solid waste. Water Science and Technology 48, 913. Hawkes, F.R., Guwy, A.J., Rozzi, A.G., Hawkes, D.L., 1993. A new instrument for online measurement of bicarbonate alkalinity. Water Research 27, 167170. HMSO, 1978. The Measurement of Electrical Conductivity and the Laboratory Determination of the pH Value of Natural, Treated and Waste Waters. Her Majestys Stationery Ofce, London. Holm-Nielsen, J.B., Lomberg, C.J., Oleskowicz-Popiel, P., Esbensen, K.H., 2008. Online near infrared monitoring of glycerol-boosted anaerobic digestion processes: Evaluation of process analytical technologies. Biotechnology and Bioengineering 99, 302313.

4090

A.J. Ward et al. / Bioresource Technology 102 (2011) 40834090 Nges, I.A., Liu, J., 2010. Effects of solid retention time on anaerobic digestion of dewatered-sewage sludge in mesophilic and thermophilic conditions. Renewable Energy 35 (10), 22002206. Spanjers, H., Bouvier, J.C., Steenweg, P., Bisschops, I., van Gils, W., Versprille, B., 2006. Implementation of in-line infrared monitor in full-scale anaerobic digestion process. Water Science and Technology 53, 55 61. Steyer, J.P., Bouvier, J.C., Conte, T., Gras, P., Harmand, J., Delgenes, J.P., 2002. On-line measurements of COD, TOC, VFA, total and partial alkalinity in anaerobic digestion processes using infrared spectroscopy. Water Science and Technology 45, 133138. Van Soest, P.J., Robertson, J.B., Lewis, B.A., 1991. Methods for dietary ber, neutral detergent ber, and nonstarch polysaccharides in relation to animal production. Journal of Dairy Science 74, 35833597. Viscarra Rossel, R.A., Walvoort, D.J.J., McBratney, A.B., Janik, L.J., Skjemstad, J.O., 2006. Visible, near infrared, mid infrared or combined diffuse reectance spectroscopy for simultaneous assessment of various soil properties. Geoderma 131, 5975. Ward, A.J., Hobbs, P.J., Holliman, P.J., Jones, D.L., 2008. Optimisation of the anaerobic digestion of agricultural resources. Bioresource Technology 99, 79287940. Williams, P.C., 2001. Implementation of near-infrared technology. In: Williams, P.C., Norris, K. (Eds.), Near-Infrared Technology in the Agricultural and Food Industries, Second ed. American Association of Cereal Chemists Inc., St. Paul, Minnesota, USA, pp. 145169.

Jacobi, H.F., Moschner, C.R., Hartung, E., 2009. Use of near infrared spectroscopy in monitoring of volatile fatty acids in anaerobic digestion. Water Science and Technology 60 (2), 339346. Jantsch, T.G., Mattiasson, B., 2003. A simple spectrophotometric method based on pH-indications for monitoring partial and total alkalinity in anaerobic processes. Environmental Technology 24, 10611067. Jantsch, T.G., Mattiasson, B., 2004. An automated spectrophotometric system for monitoring buffer capacity in anaerobic digestion processes. Water Research 38, 36453650. Kyazze, G., Dinsdale, R., Guwy, A.J., Hawkes, F.R., Premier, G.C., Hawkes, D.L., 2006. Performance characteristics of a two-stage dark fermentative system producing hydrogen and methane continuously. Biotechnology and Bioengineering 97, 759770. McCarty, G.W., Reeves, J.B., Reeves, V.B., Follett, R.F., Kimble, J.M., 2002. Midinfrared and near-infrared diffuse reectance spectroscopy for soil carbon measurement. Soil Science Society of America Journal 66, 640646. Mendez-Acosta, H.O., Snell-Castro, R., Alcarez-Gonzalez, V., Gonzalez-Alvarez, V., Pelayo-Ortiz, C., 2010. Anaerobic treatment of Tequila vinasses in a CSTR-type digester. Biodegradation 21, 357363. Moreira, M.B., Ratusznei, S.M., Rodrigues, J.A.D., Zaiat, M., Foresti, E., 2008. Inuence of organic shock loads in an ASBBR treating synthetic wastewater with different concentration levels. Bioresource Technology 99, 32563266. Nain, M.Z., Jawed, M., 2006. Performance of anaerobic reactors at low organic load subjected to sudden change in feed substrate types. Journal of Chemical Technology and Biotechnology 81, 958965.