Lab 9: Redox Reactions in Flooded Soils

Tina Gupta

Stockton University
Abstract

Redoximorphic features are associated with anaerobic conditions and formed by the oxidation of

iron and manganese oxides in soil. This study was conducted to analyze and predict evidence of

reduction reactions. Five different soil samples were used in seven flask with different periods

of saturation. Reduced iron, nitrate, manganese and hydrogen sulfide were measured using test

strips. Redox potential was measured using a Pt-electrode. Iron sulfide was determined by

swirling each flask and looking for precipitate. As a result, redox potential rates decreased,

hydrogen and iron sulfide was found in the longer saturated flasks and nitrate concentrations

were found in the recently saturated flasks. Reduced iron and manganese were mainly present

around the third week of saturated soil flask. Predictions for the mostly coincided with results.

Introduction

Redox reactions are a group of reactions that are focused on the transfer of electrons

between species. Similar to acid-base reactions, redox reactions are a paired set. The oxidation

alone and the reduction alone are each called a half-reaction (Anon 2017). Oxidation refers to

the loss of electrons while reduction is the gain of electrons (Anon 2017).

Redox reactions include all chemical reactions in which atoms have their oxidation state

changed. Even though oxidation reactions are commonly associated with the formation of

oxides from oxygen molecules it is not necessary for oxygen to be present in these reactions

because other chemical species can serve the same function (Inglett et al. 2005). The tendency

for electrons to be transferred can be measured by redox potential. A low reduction potential

will use weaker electron acceptors and tend to lose electrons to new species. As oxygen levels

are lowered, hydrogen sulfide appears from the microbial breakdown of organic matter. It is a

colorless gas which will produce an odor of rotten eggs. Some of the hydrogen sulfide will react
with metal ions in water to produce metal sulfides such as iron sulfide (Favre 2002). When both

the reduced sulfur and reduced iron are present they form a black precipitate in soil. The

increase of reduced manganese is expected but faster than reduced iron. To form low oxygen

conditions in soil, soil must be saturated. A series of 7 flasks of soil that have been flooded for

varying time periods up to 4 weeks will be observed to determine evidence of reduction

reactions. It is predicted that hydrogen sulfide, and reduced iron and manganese levels will rise

in the flasks that have been saturated for a longer time while nitrate will be present in the

recently saturated soils. Redox potential is expected to decrease in the soils that have been

saturated for longer periods.

Methods

A soil sample was taken from the Stockton University farm in Galloway, NJ. Each Erlenmeyer

flask contained 50 grams of soil followed by 0.5 grams of gypsum, 1 gram of dried alfalfa and

150 mL of water. The alfalfa was used as a nitrogen fixing bacteria. Gypsum offers an available

form of sulfate sulfur that binds with the Mg. The sulfate that is taken up by plants releases

associated oxygen which is a source of oxygen to plant roots. They were then mixed by swirling

the flask a few times and a strip of lead-acetate paper was placed in the flask secured by a

stopper. The Hydrogen sulfide gas reacts with the lead strip forming lead sulfide. A gas lock was

placed on the flask to allow Co2 to escape and prevent oxygen from diffusing in. A full 60 ml

Nalgene bottle of water was attached to the gas lock. The flask was covered with aluminum foil

to exclude light and reduce algae growth. These steps were repeated to the next flask, flask six

the following week flask 5. Flask four was prepared on the fourth week and flask three, two and

one were prepared the fifth week. Flask 3 was prepared 3 days before flask 2 and flask 2 was

prepared 1 day before flask 1. This process was repeated with four other soil samples in the New
Jersey area. After the duration of the five weeks, Hydrogen sulfide was measured using the lead

acetate strip. The flasks were swirled to check for evidence of iron sulfide precipitate. Redox

potential was then measured by placing Pt-electrode into the flask and then 199 was added to that

number to get the Eh. Indicator strips were used to measure nitrate, reduced iron and manganese.

Results

(Table 1) includes data on the color of the lead acetate paper, formation of Iron Sulfide and the

measured redox potential in mV for all five samples in the seven flask. For flask one, the most

recently saturated flask, the lead acetate paper stayed white for all five samples and there was no

indication of iron sulfide formed. The range for the redox potential varied but generally had a

high number. For flask two, lead acetate paper still stayed white and there was still no formation

of iron sulfide. The redox potential had a general lower average compared that of flask one. In

flask three we began to see some color forming such as a partial grey color on the lead acetate

paper for most of the samples. There still was no formation of iron sulfide and the trend of a

lower redox potential continues. Flask five shows a more defined gray color on the lead acetate

paper and formation of iron sulfide is present in some of the samples. The redox potential rates

are all negative at this point. By flask seven all the lead acetate papers are black and iron sulfide

has formed. (Figure 1) represents the concentration of nitrate in each of the flasks for each

sample type. For all the samples, the highest level of nitrate is in flask one or two and then flasks

4 to seven indicate no nitrate. (Figure 2) shows reduced iron concentrations. It seems that for

each sample there is no reduced iron at first but then there is a sudden increase and then

decrease. (Figure 3) indicates reduced manganese concentrations. A similar trend compared to

iron occurs here where initially there are low levels of reduced manganese then an increase with

time and then slowly decrease.

Discussion

From the data collected, redox potential rate as predicted, decreased in the soils that were

saturated for a longer time. Depending on soil characteristics, redox potential will usually

decrease with time (Inglett et al. 2005). Negative numbers represent high electron activity and

intense anaerobic conditions while positive values represent low electron activity. As for reduced

iron and manganese, it would be false to claim that both increased during longer periods of

saturation.

When oxygen availability becomes limited, bacteria uses other compounds as electron acceptors

to maintain their metabolism including oxidized forms of Nitrogen Iron and Manganese. In

recently flooded soils, nitrous oxide can be presented as a result of denitrification (Tiner 1999).

Which explains the presence of nitrate levels in flask 1, or the recently saturated soils. After this

process, selected microbes reduce manganese from Mn IV to the reduced form Mn III (Mejia et

al. 2016). Iron is then reduced from Fe III to Fe II (Favre 2002). This reduction process

continues with microbes reducing the sulfates and carbon and producing hydrogen sulfide which

forms typically in a 1-2 month period of saturation (Lal 2006). We know the hydrogen sulfide

has formed because of the way it reacts with the lead acetate paper turning it black in flask 7.

Conclusion

This study was conducted to analyze evidence of reduction reactions. It was proposed that redox

potential would decrease which it did. Hydrogen sulfide was found in the longer period of

saturation flask which could have been due to the microbes that reduce sulfates and carbon

leaving hydrogen sulfide. Nitrate concentrations were primarily found in the flasks that were

saturated for the least amount of time. Reduced Iron and Manganese was recorded around flask
five but decreased or disappeared in the flasks that followed. Thus, generally reduction reactions

corresponded effectually to predictions.

 REFERENCES

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Inglett, P. w., K. r. Reddy, and R. Corstanje. 2005. “Anaerobic Soils.” Encyclopedia of Soils in

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