Bioresource Technology 100 (2009) 471474

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

Short Communication

Simple and rapid methods to evaluate methane potential and biomass yield for a range of mixed solid wastes
P. Shanmugam *, N.J. Horan
Public Health and Environmental Engineering, School of Civil Engineering, University of Leeds, Leeds, LS2 9JT, UK

a r t i c l e

i n f o

a b s t r a c t
This paper describes rapid techniques to evaluate the methane potential and biomass yield of solid wastes. A number of solid wastes were mixed to provide a range of C:N ratios. Empirical formulae were calculated for each waste based on the results of chemical analysis and these formulae were used to estimate the COD equivalent and stoichiometric methane potential (SMP). The actual COD and biochemical methane potential (BMP) were determined experimentally for each waste and for both parameters there was a good agreement between the empirical and experimental values. The potential of adenosine triphosphate (ATP) to act as an indicator of biomass yield (mg VSS mg1 COD removed) was determined during the anaerobic digestion process. The biomass yield determined from ATP analysis was in the range 0.010.25 mg VSS mg1 COD removed which corroborated well with previously reported studies. Empirical formula based SMP together with ATP measurement were shown to provide rapid methods to replace or augment the traditional BMP and VSS measurements and are useful for evaluating the bioenergy and biomass potential of solid wastes for anaerobic digestion. 2008 Elsevier Ltd. All rights reserved.

Article history: Received 13 December 2007 Received in revised form 30 May 2008 Accepted 5 June 2008 Available online 9 August 2008 Keywords: Empirical formula Biochemical/stoichiometric methane potential C/N ratio Adenosine tri-phosphate Biomass yield

1. Introduction The techniques used in UK for disposing of solid wastes are changing rapidly and largely as a result of the Landll Directive, biodegradable waste is now being diverted away from landll. This Directive requires that by July 2010 biodegradable municipal waste (BMW) going to landll shall be reduced to 75% of its 1995 tonnage, rising to 35% of this by 2020 (Defra, 2007). Diversion is also being aided by increases in land ll tax, currently around 22/T. Other drivers that contribute to this change include climate change and the potential scal rewards from the Renewable Obligation Certicates (ROCs). As a result a large tonnage of biodegradable waste stream previously sent to landll now requires dealing with by other means. The opportunity to convert this waste to energy is obviously an attractive one and current technologies for achieving energy from waste focus on biological and thermal options. The biological route is essentially the application of anaerobic digestion (AD) which is able to reduce the amount of volatile solids in a waste feed by up to 70%, whilst at the same time generating a source of renewable energy as methane. The Department of Environment, Food and Rural affairs (Defra, 2007), has estimated that the potential annual market for anaerobic digestion is 400

* Corresponding author. Address: Environmental Engineering, Central Leather Research Institute (CSIR), Adyar, Chennai 600 020, India. Tel.: +91 44 24911386. E-mail addresses: pashanmugam@yahoo.com (P. Shanmugam), n.j.horan@ leeds.ac.uk (N.J. Horan). 0960-8524/$ - see front matter 2008 Elsevier Ltd. All rights reserved. doi:10.1016/j.biortech.2008.06.027

million. In addition AD currently attracts 2 ROCs with a value of 30/MWh. However, AD is a capital intensive project and in order to undertake a basic process economic feasibility study, it is important to have an indication of the likely methane yield available from the digester feedstock. A number of techniques are available to provide this information including the biochemical methane potential (BMP), dynamic respiration rate (DR4), and the COD test (Owen et al., 1979; Environment Agency, 2005). In addition, a number of indicative ratios can be used such as the soluble COD to volatile organic solids ratio. Of these, the BMP test is the most popular but, as with any simple batch test that is intended to provide information on the likely performance of a full-scale continuously operated process, the results of the BMP require interpretation with caution. In particular, the BMP test is conducted over a period of 28 days, whereas conventional AD rarely operates at retention times above 15 days. Many researchers have searched for feasible alternatives to the BMP test, but with limited success and consequently rather than attempting to search a replacement, it is perhaps more appropriate to develop additional protocols to complement the results from the BMP and aid in its interpretation (Shelton and Tiedje, 1984; Mendez and Lema, 1993). This paper intends to evaluate two such complementary test procedures. The rst of these involves determination of the empirical formula of biomass that can be used for stoichiometric product estimation (Erickson, 1978, 1980). The empirical formula developed for activated sludge from aerobic biological treatment was C5H7NO2 (Rittmann and

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McCarty, 2001; Tchobanoglous et al., 2005) and routinely used as basic design rule to calculate the oxygen requirement for BOD removal. In a similar way Rittmann and McCarty (2001) and Hansen (2005) have proposed that stoichiometric methane potential (SMP) determined from the assessment of empirical formula based organic solid wastes will provide information on the energy balance for fermentation processes. Even with the SMP test, information is not available as to the behaviour of bacteria during the AD process and those conditions which provide an optimum environment for their growth. The direct measurement of volatile suspended solids (VSS) cannot be applied in solid wastes digesters as it cannot distinguish between bacteria and suspended organic solid wastes particles and so there is a need for an alternative method of VSS measurement for solid waste digesters to predict biokinetic design constants. The quantication of adenosine triphosphate (ATP) has been used by a number of researchers as an anaerobic cell viability assay method (Chung and Neetheling, 1988; Yu et al., 2002; Kim et al., 2007). The ratio of ATP to VSS (Hwang and Hansen, 1998) has also been used to assess both cell viability and biomass yield, thus providing important information on the behaviour of the organisms during the AD process. Hence, this present study investigates the SMP and ATP analysis as complementary tests to the BMP and VSS for evaluating the suitability of a range of organic solid wastes as substrates for AD.

trolled gas opening valves. The effective volume maintained was 400 ml and the gas phase was 100 mL. Nutrient medium was prepared using a modication to the method described by Owen et al. (1979) and added at 1 mL per litre of reactor volume. An internal temperature of 35 C was maintained by incubating the reactors in a temperature controlled mechanical shaker. Samples were mixed at 140 rpm for a 15 minutes period followed by 15 minutes with no shaking. The quantity of biogas produced was measured by connecting the gas opening valve on the reactor to the inlet tube of a hermetically sealed, water displacement aspirator bottles lled with 5% NaOH to scrub CO2. A measuring cylinder at outlet of the aspirator bottles collected displaced water which measures CH4 at atmospheric temperatures and pressure (Nm L CH4. day1). The initial and nal characterisation of 500 mL bottles was taken for mass balance analysis and the serum bottle samples were used for anaerobic process evaluation analysis. The BMP of seed sludge was simultaneously carried out in control reactors, and subtracted from the VS and gas yield with solid waste and the BMP yield of solid waste was calculated as Nm L gm1 VS removed. The contents of the serum bottle were withdrawn periodically and centrifuged at 6000 rpm for 1 h and the supernatant was collected for product analysis of total alkalinity (T.Alk), volatile fatty acids (VFA), and ammoniacal nitrogen (NH3N). 2.4. ATPcell viability and biomass yield Analysis of ATP was based on quantication of luminescence released from the reaction of luciferase with ATP (Chung and Neethling, 1989). Samples were diluted with 20 mM Tris-EDTA at pH 7.75, boiled for 30 min and equilibrated to room temperature. Supernatant ATP was recovered by centrifugation at 6000 rpm for 5 minutes. The luminescence reaction of sample ATP and luciferase was measured as a Relative Luminescence Unit (RLU) in a Promega GloMaxTM 20/20 model luminometer, following addition of back titre luciferase supplied by Promega. A calibration graph was prepared from the RLU at varying ATP concentrations with luciferase to quantify the sample ATP. The COD and VSS content over time was calculated using 1.4 mg COD mg1 VS and 250 mg VSS mg1 ATP (Hwang and Hansen, 1998). This was then used to calculate biomass yield as mg VSS mg1 COD removed.

2. Methods 2.1. Characterisation of solid wastes A number of solid waste sources were used in this study. Municipal solids waste (MSW) was collected in bulk and autoclaved at 130 C to permit storage without deterioration. Leather eshing (LF) and primary chemically treated sludge (LS) were collected from the tannery efuent treatment plant at Holmes Hall tannery in Hull. The leather eshing was minced and homogenised with a commercial blender to 6 mm diameter before feeding to the digester. Wastewater treatment sludge included primary sedimentation tank sludge (PST), mixed liquor from a sequencing batch reactor (SBR), and surplus activated sludge (ASP) were collected from the Knostrop wastewater treatment plant at Leeds. The characterisation of total solids (TS), and volatile solids (VS) were carried out using Standard Methods (APHA, 1998). Elemental analysis of carbon, hydrogen, nitrogen and sulphur was undertaken using CHNS analysers Model Thermo Flash EA 1112 series. Samples were oven dried at 103 C and then combusted at 1800 C in the CHNS analyser in a steam of helium with measured amount of oxygen. This produces N2, CO2, H2O, and SO2 which are then separated and quantied by gas chromatography using a 5 mm diameter steel (length of 2 m) packed column, helium carrier gas with a ow rate of 40 mL/min, detected with a Propack model TCD. 2.2. Development of empirical formula for stoichiometric methane potential (SMP) From the calculated values of C, H, N, and O an empirical formula was computed for each solid waste following the procedure developed by Rittmann and McCarty (2001) and Hansen (2005). This formula was used to determine the COD equivalent of which permitted the calculation of the stoichiometric methane potential (SMP). 2.3. Biochemical methane potential (BMP) assessment The BMP was determined in anaerobic batch reactors of 500 ml capacity Duran bottles with hermetically sealed stoppers and con-

3. Results and discussion 3.1. Characterisation of a range of raw solid wastes The important parameters for characterising the suitability of a waste for AD are carbon to nitrogen ratio (C/N) and VS content, and the microbiological performance parameters such as BMP and biomass yield are necessary for reactor design estimations. A range of solid waste types with differing characteristic were selected for this study (1) with the volatile organic solids ranging from 60 to 75% and the C:N 3.2 to 21.6. The LS had the highest organic content, whereas the LF had the highest nitrogen content and thus a lower C/N ratio. Primary sludge had the highest ash content and this is likely to be a result of the presence of silt, clay and sand particles. 3.2. Empirical formulae, stoichiometric COD and methane potential (SMP) Based on the chemical analysis provided in Table 1, the empirical formula of each waste was calculated (Table 2). The calculated empirical formula was then used to determine the theoretical COD (CODe) of the waste. When expressed in terms of the measured volatile solids this gave an average value of 1.4 g CODe g1 VS, a value that is typical of such wastes, for instance Han et al. (2005)

P. Shanmugam, N.J. Horan / Bioresource Technology 100 (2009) 471474 Table 1 Characterisation of range of solid wastes Parameters 1. 2. 3. 4. 5. 6. 7. 8. Total dry solids (%) Volatile solids (% of TS) Carbon (% of TS) Hydrogen (% of TS) Nitrogen (% of TS) Oxygen (% of TS) Sulphur (% of TS) Ash content (% of TS) Leather eshings 21.9 81.3 35.5 9.4 11.2 25.3 0.6 18.7 ASP sludge 0.9 70.3 37.6 5.7 7.0 20.1 0.5 29.7 SBR sludge 0.4 62.0 28.3 5.1 4.2 24.5 0.8 38.0 Leather sludge 21.5 81.5 39.9 6.4 3.5 31.6 1.3 18.5 PST sludge 3.8 60.3 28.7 4.6 2.4 24.7 0.5 39.7

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MSW 41.7 63.0 27.5 3.8 1.3 30.5 0.4 37.0

Table 2 Empirical formula, and anaerobic process evaluation for a range of solid wastes Solid waste Leather eshing ASP sludge SBR sludge Chemical sludge PST sludge MSW Empirical formula C4H11NO2 C6H11NO2 C8H17NO5 C13H26NO8 C14H26NO9 C25H41NO21 C:N 3.2 5.4 6.7 11.4 14.0 21.6 VFA (mg L1) 3540 4147 4266 2722 3490 4371 T.Alk. (mg L1) 9163 6039 7997 7477 5703 6821 NH3N (mg L1) 1296 241 466 999 305 414 VSSe (g g1 COD eliminated) 0.09 0.14 0.14 0.01 0.12 0.25

reported values of 1.1 and Rittmann and McCarty (2001) observed 1.4. Good correlation was also observed between the CODe determined from the empirical equations and that calculated experimentally. Direct measurement of the COD of a solid waste is often thought to produce erroneous results and thus the volatile solids are generally used to quantify the organic strength of solid wastes (Han et al. 2005; Lin et al., 1999). However, COD data is required for reactor design to estimate the biokinetic yield. Consequently COD estimation based on empirical formula provides an attractive alternative to the experimental COD for design purposes. 3.3. Comparison of SMP and BMP test The BMP was measured using the same range of solid wastes under controlled conditions of nutrient addition, heating and stirring. The measured BMP ranged from 0.36 Nm L g1 VS for the chemical sludge to 0.52 Nm L CH4 g1 VS for the ASP (Table. 2). These values are typical of those observed by others, for instance Lin et al. (1999) reported a BMP of 0.35 Nm L CH4 g1 of CODe (equivalent to 0.318 Nm L g1 VS) for chemically pretreated ASP sludge. A comparison of the SMP and BMP for these solid wastes gave a good linear relationship (Table 2). The SMP of LF, PST, SBR, and LS were 0.58, 0.53, 0.51, and 0.56 Nm L g1 VS whereas; the BMP was recorded as 0.49, 0.38, 0.38 and 0.36 Nm L g1 VS, respectively. The observed SMP and BMP of ASP sludge was 0.61 and 0.52 Nm L g1 VS, in line with the ndings of Gosset and Belser (1982) and Wook and Hwang (2000) who reported the BMP of ASP sludge as 0.360.54. In a similar way, the SMP and BMP of the MSW was 0.40 and 0.36 Nm L CH4 g1 VS and a number of authors report similar ndings, for example Davidsson et al. (2007) who reported that MSW has a BMP of 0.30.4 Nm L g1 VS. Thus the SMP, which can be undertaken within 24 h, provides a more rapid alternative to the 28 days BMP test. It has the additional advantage that it provides information as to the likely COD of the waste under test and such information can be used for further biokinetic evaluation. 3.4. Evaluation of factors affecting full-scale digester performance The SMP and CODe offer rapid procedures to estimate the potential methane yield from a waste. However, they give no indication as to how the waste might degrade in a full-scale digester. Many other factors inuence actual digester performance. For

instance poor VS removal efciency and low biogas yield are often associated with low C/N, high NH3N, high VFA and low digester buffering capacity (Haandel and Lettinga, 1994; Anderson et al., 2003). In addition, the concentration of ATP is known to indicate both the cell viability and the metabolic status of a microorganism and it has been used to predict biomass levels in anaerobic digestion (Chu et al., 2001; Yu et al., 2002; Kim et al., 2007). In order to establish the potential of ATP as a simple performance monitor, ATP concentration was measured over the duration of the BMP test, together with a number of physical parameters that indicate reduced performance. By monitoring a range of solid waste types with differing C/N ratios it was hoped that a clear picture would emerge as to the status of ATP, as an indicator of reactor stability and performance. C/N ratio was selected as the variable parameter because it determines both the NH3N and VFA concentration observed in the digester. Increasing NH3N helps to raise the pH, whereas by contrast an acid pH is governed by VFAs which neu2 tralise HCO and acetate ions. However ammonia can be 3 , CO3 toxic: Callaghan et al. (2000) and Salminen and Rintala (2002) have reported the tolerance level of NH3 in anaerobic digesters acclimatised to treat high protein wastes as 11.6 g L1 and 6.0 g L1, respectively. Of the six solid wastes evaluated, the cumulative biogas yield after 28 days was highest for MSW and the lowest for leather eshing, in line with the C:N ratios for these wastes (Fig. 1 and Table 2). The maximum NH3N concentration of 1296 mg L1 was also recorded for the leather eshing but this is well below the concentration reported as inhibitory. Alkalinity was in the range of 3800 9700 mg L1 and the maximum alkalinity was coincident with high NH3N levels (Table 2). The VFA was observed to peak at around 8 days for SBR, ASP and MSW wastes, whereas for LF it was as late as 20 days suggesting that a much longer digester retention time would be required for this waste (Fig. 2). The late decrease in VFA was also coincided with high alkalinity. ATP was measured at a concentration in the range of 2.018.43 mg L1 and similar values have been reported by others, for instance Hwang and Hansen (1998), Yu et al. (2002), and Chen (2004). The ATP level for LF, ASP, SBR, LS, PST, and MSW were 2.01, 4.21, 3.22, 2.12, 3.22, and 8.43, respectively, and observed as linear with respect to cumulative methane yield measured during the BMP test (Table 2), thus demonstrating the potential value of this parameter. Standard BMP test can be used to estimate biokinetics design constants. Future work

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3000 2500 Biogas (N mL) 2000 1500 1000 500 0 0 10 20 30 Days Elapsed 40
SBR ASP PST Chemical Slud Leather Fl MSW

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Fig. 1. Cumulative biogas evolution with elapsed time.

9000 8000 7000 VFA (mg/L) 6000 5000 4000 3000 2000 1000 0 0

SBR Sludge ASP sludge PST Sludge Chemical Sludge MSW LF

10 20 30 Elapsed time (Days)

40

Fig. 2. The volatile fatty acids (VFA) levels at elapsed time during the BMP test.

will therefore focus on correlating the level of ATP attained in the BMP test with the biokinetic constants of a continuously fed system treating the same waste type. 4. Conclusion For all the solid waste examined, there was a good corroboration observed between SMP and BMP, CODe and experimental COD. The wastes with higher C:N ratio produced methane more rapidly and also provides a guide to the likely operating retention time of a continuously fed digester. The ATP measured was indicative of both volatile solids removal, amount of CH4 and VSS (bacteria) produced from the wastes. Thus, the combined application of empirical formulae, SMP and ATP offers the potential to augment BMP and VSS tests to estimate both methane yields and biokinetic design constants for solid wastes anaerobic digesters. Acknowledgements The authors would like to acknowledge the help of Mr. Nick Waudby, Holme Hall Tannery, Hull (UK) for providing samples of leather eshing and chemically treated sludge.