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In Situ Aerobic Biodegradation In Soil Contact Using The Standard Test

Method of ASTM D 5988-12
To cite this article: S.M. Al-Salem et al 2019 IOP Conf. Ser.: Earth Environ. Sci. 291 012011

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2019 3rd International Conference on Energy and Environmental Science IOP Publishing
IOP Conf. Series: Earth and Environmental Science 291 (2019) 012011 doi:10.1088/1755-1315/291/1/012011

In Situ Aerobic Biodegradation In Soil Contact Using The

Standard Test Method of ASTM D 5988-12

S.M. Al-Salem, A. Al-Nasser, H. Sultan and H. Karam

Environment and Life Sciences Research Centre, Kuwait Institute for Scientific
Research, P.O. Box: 24885, Safat 1319, Kuwait.

Abstract. With an over growing problem of plastic waste accumulation, Kuwait faces a serious
threat to its natural resources. Latest estimates show that plastic solid waste (PSW) is generated
at an alarming rate of over 200 ktpa, making up over 16% of the total municipal waste load. In
this paper, the standards test method of ASTM D 5988-12 for determining the degree and rate
of biodegradation of plastics in contact with soil was applied to three commercial plastic
products sourced locally. The plastic samples are claimed to be of biodegradable nature by the
manufacturer and the converter. The test method was applied in situ using analytical
desiccators and have proven to be a good indicator to biodegradable measurements. In addition
to technical control and reference tests, the test method was also applied on the natural soil
secured from the field simulating the largest operating landfill site. The test has proven to be
reliable and has produced repeatable results. The biodegradation extent was noted to be very
rapid and can be described by comparison to other works as accelerated. This was attributed to
both what is described by this work as external and internal factors of the polymer-soil matrix.
The former can be noted as environment and soil characteristics, pH level and presence of
contaminants in the biota. On the other hand, the latter can be described as type of soil (mixture)
ratio, polymer surface area and carbon dioxide (CO2) evolution kinetics which were noted to be
of a first order with respect to biodegradation time and dissolution.

1. Introduction
The rapid development in plastic articles and their properties has made their management post disposal
an alarming threat to the natural environment. Polymeric articles constitute a number of chemicals and
additives that are used to enhance their properties [1]. However, biodegradation has made it possible
to alter these hydrocarbon (HC) chains and make them susceptible to chemical bonds’ cleavage [2-3].
Therefore, it is deemed necessary to start having test procedures and standards that can govern the use
and assessment of biodegradable polymers. Therefore, devising a standard test methodology for the
determination of biodegradability extent is noted to be of essential need nowadays. The American
Society for Testing & Materials (ASTM) has developed and reported such a test method (D 5988-12).
The test determines the degree and rate of biodegradation under controlled laboratory conditions in
contact with soil. This method was chosen in our work due to the fact that it reports the biodegradation
extent and rate, in addition to, simulating the actual microorganism attack by the local environment.
The purpose of this communication is to report the study results of three commercially produced and
converted (claimed to be) biodegradable plastic film products. The reliability of the test method is
discussed, in addition to the results obtained from the analytical work, which is the first of its kind in
the Middle-East region. The test is also noted to provide very essential information on the
biodegradation of local soil secured from Kuwait’s arid land (Al-Maqwa area) which also simulates
the activities of the largest operating landfill site in the country (seventh ring road - south).

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2019 3rd International Conference on Energy and Environmental Science IOP Publishing
IOP Conf. Series: Earth and Environmental Science 291 (2019) 012011 doi:10.1088/1755-1315/291/1/012011

2. Experimental
Three types of plastic bags used in the Kuwaiti market were secured from local dealers with their main
characteristics and sample dimensions depicted elsewhere [5,6]. The samples acquired were presumed
to be environmentally friendly and biodegradable, which were secured from various sources within the
state. The samples were cut into standard testing dimension specimens accepted by international
protocols for thin plastic film characterization as described previously [1,5]. The determination of the
biodegradability extent was conducted in this work under the presumption that the samples were
biodegradable in nature with suitable content of pro-degradants (or oxo-biodegradable) as declared by
the company’s marketing these polymeric products. The biodegradation extent was investigated via
aerobic bacteria in contact with soil under controlled conditions [6]. The standard testing procedures
of ASTM D 5998 [7] were followed to determine the rate of aerobic biodegradation of plastic
materials in soil. Natural soil (particle size < 2 mm) was used after reclaiming it from a local dealer in
Kuwait in the amount of 50 kg. The soil originates from the Maqwa’a area which is the closest to the
largest operating landfill site in Kuwait, dug up from 1 to 3 m (top soil) stored in a cold room storage
with a maintained temperature between 1 to 3oC. Compost (sheep manure) sourced from local dealers
was also used in this work. Potato starch (MSDS CAS no. 009005-84-9) purchased from Loba Chemie
Company was used for positive control experimental runs. The soil was aerated with the compost in
laboratory conditions and mixed in a 1:25 ratio (e.g. 18:450 g compost/g soil) with a moisture content
of 80-100% maintained throughout the full duration of the experiment by adding distilled water. The
moisture holding capacity (MHC) of the soil was determined initially after drying it in an oven for 24
h at 105oC and was estimated to be 0.6%. Each experimental run consisted of six air tight 2 litre
desiccators. The first was denoted as blank (control) which contained the soil. A technical control
experimental run to make sure that the experimental setup was free from leakage, was also executed.
A positive control experiment was conducted in conjunction with the first two containing the starch
and soil. The remaining three desiccators contained the samples studied. Plastic bags sample size were
used as described in the experimental protocol of ASTM D 5998 [7] and Rudeekit et al.[8] which were
2x2 cm2. The amount of 200 mg for each type and positive control (starch) sample were used. The
carbon (C) to nitrogen (N) ratio was maintained in the experimental setups after the addition of
Ammonium Phosphate ((NH4)2HPO4) (4.72 g/l, Sigma Aldrich Company reagent >98%) and
maintaining the pH 8.2 [6]. The C:N ratio was maintained at 10:1 after addition to the soil.
The test method described aims at the determination of aerobic biodegradability, obtaining soil as a
matrix and source of inoculum, as well as, estimating the degree of biodegradability [7]. To this end,
both barium hydroxide (Ba(OH)2) and deionized water (DI) were used within each experimental setup,
placed inside each of the desiccators used. The Ba(OH)2 solution was prepared by dissolving 4.0 g
anhydrous Ba(OH)2 (Aldrich Company, technical grade 95%) in distilled water and confirmed by
manual titration. The solution was sealed and stored to prevent absorption of CO 2 from the air. The
amount of 100 ml of 0.025 N Ba(OH)2 and 50 ml of distilled water were kept constantly throughout
the experiments within each of the desiccators used on the perforated plate inside the vessel. All tested
were conducted in dark room in laboratory conditions. Manual titration was conducted in a frequency
of 3 days for the first three weeks and once every three weeks thereafter as recommended previously
by Gomez and Michael [9] and ASTM D5998 [7]. The titration aims at determining the accumulative
carbon dioxide (CO2) evolution, conducted using hydrochloric acid (HCl, 0.05 N) and Ba(OH)2 to a
phenolphthalein end-point. The theoretical total carbon (TC) was determined for each type of plastic
bags and starch using dynamic flash combustion using modified Dumas method [10] in a Thermo-
Fisher Scientific (Flash 2000 Model) CHNS-O analyser. Samples were conducted in triplicates for
quality assurance. The theoretical CO2 quantity was calculated using reaction stoichiometry thus

44  Y (1)
CO2 (th.) 
12

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2019 3rd International Conference on Energy and Environmental Science IOP Publishing
IOP Conf. Series: Earth and Environmental Science 291 (2019) 012011 doi:10.1088/1755-1315/291/1/012011

where Y is the net total theoretical carbon (C) charged to each vessel (mg), and CO2(th.) is the
theoretical CO2 available (mg). The amount of net CO2 produced is calculated by subtracting the
titration readings from the test material titration thus

Z n  Zb  Zt (2)

where Zn is the calculated amount of HCl needed to titrate the CO2 generated solely from the test
material (ml), Zb is the amount of HCl used to titrate the blank vessels with soil only (ml) which is
termed as technical control vessel in this work, and Zt is amount of HCl used to titrate the vessels
containing the soil plus test material (ml). The amount of CO2 produced is calculated as per the
formula given below after accounting for the Normality of HCl [7]

1.1 0.05 N  Z n  44  (3)

CO 2 (Produced) 
2

The percentage of CO2 evolution (E) is determined as per the following equations, which on the other
hand is used to estimate the standard deviation (se) of the experimental runs and biodegradation extent
thereafter.

CO 2 (Produced) (4)
E (%)  100
CO 2 (th.)

se  s 2
test /n1   s 2blank /n 2  100/C i (5)

where n1 and n2 are the number of replicate test material and blank vessels, respectively, s is the
average standard deviation of the total gaseous carbon produced (mg), and C i is the amount of carbon
(mg) initially present in the vessel.

3. Results and discussion

As an initial trail and in order to assure that the soil material used will accommodate the ASTM D
5988 test method undertaken in this study, trails on the soil without the presence of samples was
conducted to assure microbial activity within the degrading environment. The method and
experimental design is similar in nature to the blank (control) experiment. The difference was in the
soil material preparation procedure and conditioning. For these initial trails, the soil was aerated for
the duration of three days and prepared in similar fashion to the previous description. The soil was
stimulated under 35oC for maximum microbial activity for the period of nine days. Throughout the
experimental efforts, including the ones dedicated for the plastic specimens, no substantial change in
the amount of distilled water was noticed. However, droplets creating a thin film was noticed around
the desiccators walls. This points towards the formation of barium carbonate (BaCO3) which
precipitates due to the reaction of the formed CO2 with the barium hydroxide (Ba(OH)2). This was
continuously avoided by gently shaking the vessels as the test method suggests. This will also
neutralize CO2 to OH- ion which is not dependent on the position of the CO32  HCO3 equilibrium
at the pH phenolphthalein endpoint [11].
Fig.1. indicates the CO2 evolution rate with time for the soil stimulated at 35oC for the nine days
monitoring time of the blank test. This verifies that the soil is compatible for conducting the test
method described in the previous section. By association, the kinetics of evolution can be described by
Michaelis-Menten mechanism in a homogenous-phase solution [11]. By comparison to the rate of CO2
evolution estimated for the actual test, it can be noted that actual test conditions facilitated more CO2
production as time has progressed and passed the 10 days mark. Therefore, the soil stimulation with
temperature can be noted to be less of a factor in the increased CO2 production

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2019 3rd International Conference on Energy and Environmental Science IOP Publishing
IOP Conf. Series: Earth and Environmental Science 291 (2019) 012011 doi:10.1088/1755-1315/291/1/012011

Figure 1. Production of carbon dioxide (CO2) as a function of time (○) measured in the initial trail
test without addition of sample with soil aeration period of three days based on the titration volume of
5 ml.

Figure 2. Biodegradation extent (%) determined for the three plastic samples tested in this work (raw
data).
The biodegradation rate was also estimated for the types of the samples studied. It was noted that the
sample thickness and possible starch content, can be major factors in the rate of biodegradation. Fig.2
shows the biodegradation extent (%) of the samples considered. It can be noticed that the samples of
type I sustained the longest period of time with biodegradation extent reaching 14 days. However,
samples of type II and III have deteriorated at higher rates. The plateau level of CO2 production
evolution in this test method and, optionally, the quantified residual test material will suggest the
degree of biodegradability of the plastic material.

4. Conclusion
The biodegradation extent using the standard test methods of ASTM D5988, was conducted on three
different types of plastic samples. The test have started using blank soil tests to investigate the biota
activity. It was noted that the test method supported soil and polymer matrix influential factors noted
as major influencers on the biodegradation rate. Two sets of factors have been noted to affect the bio-

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2019 3rd International Conference on Energy and Environmental Science IOP Publishing
IOP Conf. Series: Earth and Environmental Science 291 (2019) 012011 doi:10.1088/1755-1315/291/1/012011

testing, external and internal ones. The kinetics, with biodegradable substrate and CO2 production rate,
has also followed the Michaelis-Menten mechanism. The former can be noted as environment and soil
characteristics, pH level and presence of contaminants in the biota. On the other hand, the latter can be
described as type of soil (mixture) ratio, polymer surface area and carbon dioxide (CO2) evolution
kinetics which were noted to be of a first order with respect to biodegradation time and dissolution.

5. References
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linear low density polyethylene (LLDPE) films Mater. Des. vol. 30, no. 5, pp. 1729-1736.
[2] Al-Salem SM, Bumajdad A, Khan AR, Sharma BK, Chandrasekaran SR, Al-Turki FA, Jassem
FH, Al-Dhafeeri AT 2018 Non-isothermal Degradation Kinetics of Virgin Linear Low
Density Polyethylene (LLDPE) and Biodegradable Polymer Blends J. Polym. Res.
(Accepted).
[3] Rosa, D. S., M. A. G., Bardi, L.D.B. Machado, D.B. Dias, L.G.A. Silva and Y. Kodama 2010
Starch plasticized with glycerol from biodiesel and polypropylene blends mechanical and
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Aspects and Special Applications Wiley first edition.
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[6] Al-Salem SM, Al-Nasser A, Al-Wadi M, Sultan H, Al-Foudaree M, Karam H, Al-Dhafeeri A,
2018 Biodegradable Films Physical Properties Change With Respect To Degradation
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on Engineering for Waste and Biomass Valorisation, Prague, Czech Republic, 2nd-5th July.
[7] American Society for Testing and Materials (ASTM) 2012 Standard Test Method for
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[8] Rudeekit Y, Leejakpai T, Euaphantasate N, Kongsuwan K 2004 Comparative degradation of
biodegradable plastics aerobic microorganim using ASTM D 5338-98 (03) and ASTM D
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6. Acknowledgments
This work is funded by the Kuwait Institute for Scientific Research and Kuwait Municipality under the
grant for Project EM097C.