NOTES

1191

COLORIMETRIC DETERMINATION OF OXIDIZABLE CARBON IN ACID SOIL SOLUTIONS RICHMOND J. BARTLETT* AND DONALD S. Ross Abstract
A simple and sensitive method is described that determines, colorimetrically, oxidizable organic matter in solutions from acid soils. It relies on measuring the loss of color by a Mn(III)-pyrophosphate complex as Mn(III) becomes reduced by organic C in the presence of concentrated H2SO4. The method is applicable to 1-mL samples containing 0.08 to 4.0 MHIO! of organic C and is practically free of interferences in aerobic solutions.

developed the following equation to explain the observed stoichiometry for the formation of a hypothetical complex and the observed amount of acid needed to prevent its precipitation or dismutation: 15Na4P2O7 + 4Mn2+ + MnO4- + 23H+ _ 5Mn(Na2HP2O7)3 + 4H2O + 30 Na+ To a small test tube, add an aliquot of solution containing 0.15 to 8 fimol of organic C and enough distilled water to bring the volume to 1.0 mL. Standards can be prepared from either oxalic acid, a humic acid solution of known C content, or by simple dilution of the Mn(III) complex. Add 0.5 mL of the 10 mM Mn(III)-pyrophosphate, 0.5 mL of concentrated H2SO4, and mix thoroughly by vortexing. Allow solutions to stand about 18 h before transferring to cuvets and determining absorbance at 495 nm on a spectrophotometer. If a solution loses all color or turns yellow instead of retaining some pinkish color after adding the H2SO4, either repeat the determination, using a diluted sample, or add additional portions of H2O, Mn(III), and H2SO4 (in a 2:1:1 ratio) and multiply concentration found by the appropriate factor. Results and Discussion A standard curve, using 0.08 to 4.0 /*mol C as oxalate, typically has a range of absorbance between 0.227 and 0.030. A series of 10 standards gave the following regression equation: Mmol C = 4.636 - 20.4 absorbance R2= 1.00 . The variability is low, nine samples each of a blank and a 2 /miol C oxalate standard gave absorbance readings of 0.221 0.003 and 0.126 0.003, respectively. Because a standard oxalate C, having a valence of + 3, donates one electron to each Mn(III) reduced, the standard curve represents moles of C oxidized. An average C atom in a representative fulvic acid, C30H48O39 (Schnitzer and Khan, 1972), has a valence of +1 and might be expected to require three Mn(III) atoms if oxidized to CO2. However, results for a "standard" ash free fulvic acid preparation (Contech E.T.C., Ottawa, Canada) showed that an average C reduced a single Mn(III). That is, the Mn(III) reduced, rather than being a measure of equivalents of C oxidized, instead was a measure of the number of C atoms. Similarly with glucose (C6H12O6), 6 Mn(III) units were shown to be reduced by each molecule of glucose instead of the 24 that would be required if each glucose C were oxidized to CO2. The much greater sensitivity of the Mn(III) method than Cr(VI) methods makes comparison between them difficult in low organic soil solutions. However, results with leachates from six forested Spodosols and six pH 7 extracts using dilute KOH (a forested Udorthent A, a Udorthent Ap garden soil, a Florida Medifibrist, and three Spodosol B horizon samples) showed close correlation between values obtained with Mn(III) and a modified Mebius dichromate procedure (Nelson and Sommers, 1982). Again 1 mole of Mn(III) was reduced for each mole of C present (Fig. 1). In four additional KOH extracts of neutral or somewhat alkaline soils, all high Mn soils, the dichromate values were almost twice as high as with the Mn(III) pyrophosphate. We can speculate that the most oxi-

ION AND MANN (1946) demonstrated that a colD ored Mn(III) complex was formed when soils were shaken with phosphate buffers and especially with pyrophosphate. This complex, the color of which can be described as violet-pink with a tinge of orange, is stable in solution below pH 7. If oxidizable organic matter is present, however, the addition of strong acid results in the immediate loss of some of this color accompanying the oxidation of organic C and the reduction of Mn(III). Measuring this loss of color is the basis for the proposed method. While not as sensitive as C analyzer methods, it requires no expensive equipment and makes analysis possible on samples of limited size, such as extracted soil solutions. Procedure Care needs to be taken for proper preparation of the oxidant. To about 100 mL of distilled water in a 250-mL volumetric flask, add in order: 75 mL of 0.1 MNa4P2O7, 11.5 mL of 0.5 M H2SO4, and 5.0 mL of 0.10 M KMnO4. Mix, and immediately add with mixing 20 mL of 0.10 M MnSO4) and bring to volume. The colored solution is 10 mMMn(III)pyrophosphate and should have a pH of 4.3. It will remain clear for several weeks if kept away from bright light. We
Department of Plant and Soil Science, Univ. of Vermont, Burlington, VT 05405. Research supported by the Vermont Agric. Exp. Stn., Univ. of Vermont. This research was supported in part by grants from the Electric Power Res. Inst, contract RP2365, ALBIOS project, and by the U.S. Dep. of Energy, DE-AS05-83ER60179. Received 26 May 1987. *Corresponding author. Published in Soil Sci. Soc. Am. J. 52:1191-1192 (1988).

1192

SOIL SCI. SOC. AM. J., VOL. 52, 1988

Table 1. Examples of potentially interfering ions. Ion tested Cl- as NaCl Br as NaBr NO; as NaNO, Cr(III) as Cr(NO,),
100-1

Cone, giving a decrease of 0.005 in absorbance

42 mM 0.9 mAf 43 pM 101 i*M

Decrease in absorbance from a 1 mM solution 0.000 0.005

0.114

0.060

A NEW TECHNIQUE FOR EVALUATING THE PRESENCE OF PREFERENTIAL FLOW PATHS

IN NONSTRUCTURED SOILS
TJ

80-

H. C. VAN OMMEN,* L. W. DEKKER, R. DIJKSMA, J.

HULSHOF, AND W. H. VAN DER MOLEN

8
60-

TJ O
<D

Abstract
A new technique for determining the presence of preferential flow paths in a nonstructured soil profile is described. The presented method is easily applicable in the field, and is based on the formation of an intensely colored complex of I, with starch. Moreover, the method can be applied to all kinds of nonstructured soils, irrespective of their color. An illustration of the technique is given in case of water and solute movement in a water-repellent soil, where formation of preferential flow paths was known to occur.

c
40-

"5

E
20Mn(lll) reduced = -0.11 + 0.974 Mmol C R2=0.98

20

40

60

80

100

fj,mo\ C from Cr(VI), Mebius Method Fig. 1. Comparison of the Mn(III)-pyrophosphate method with the modified Mebius method. Samples were leachates or extracts from 12 acid soils.

ANY STUDIES CONCERNING solute movement M through the unsaturated soil profile have shown that the leaching mechanism depends upon the variability of water fluxes in the field, (e.g., Van de Pol et al, 1977; Nielsen et al., 1986; White et al., 1986). In structured soils, such as clay and peat soils, cracks and macropores may contribute an important part to this variability. Techniques to visualize these macropores and cracks, and their capacity of transporting water and solutes have been introduced by Bouma and Dekker (1978), and Bouma et al. (1978, 1979). These often involve the application of an amount of dye solution to the soil surface. The presence of the dye, which adsorbs at the walls of the cracks and macropores, can be studied, for example, by micromorphological techniques. In nonstructured soils, the phenomenon of flow through preferential paths is not linked with the presence of cracks or macropores; its occurrence can be induced by several mechanisms. The mechanism of formation of unstable wetting fronts, possibly generated by an increase of the hydraulic conductivity with depth (Raats, 1973), causes the development of preferential paths through the soil matrix. Another mechanism concerns the formation of preferential paths in a soil profile with water-repellent characteristics, as has been described by Bond (1964). An important difference between water and solute flow in structured and nonstructured soils, is the depth at which a preferential flow path develops. In structured soils it originates principally from the soil surface; in nonstructured soils, the development of a preferential flow path may take place at any depth, depending upon soil hoDep. of Land and Water Use, Agric. Univ., Nieuwe Kanaal 11, 6709 PA Wageningen, Netherlands. Contribution of the Dep. of Land and Water Use, Agric. Univ., and the Netherlands Soil Survey Inst., P.O. Box 98, 6700 AB Wageningen, Netherlands. Received 29 May 1987. *Corresponding author. Published in Soil Sci. Soc. Am. J. 52:1192-1193 (1988).

dizable organic matter was already lost from these soils, decomposed by natural Mn oxides. This is an interesting subject for further study, but in the meantime, we recommend that this method be applied with caution in alkaline extracts of soils with pH's above 6.
We must be aware that we may be observing C that

is oxidizable rather than total in such soil solutions. Although extremely rare in aerobic soil solutions, reactive redox species could interfere if present (Table 1). Nitrites, sulfides, Fe(II), and Cr(III) produce positive interferences by reducing the Mn(III). Peroxides, O free radicals, and Mn(III) and Mn(IV) complexes could produce negative interferences if present in soil solutions. Even though the oxidation of chloride to chlorine and bromide to bromine by Mn(III) is thermodynamically favorable, this reaction occurs only with high levels of chloride or bromide under the conditions of this test (Table 1). Humic polymers, proteins, and polysaccharides are readily oxidized, but a few common simple organic compounds resist oxidation in the Mn(III)-H2SO4 system, notably acetic acid and glycine. Manganese(III) differs from both chromate and permanganate in that its reduction involves only one electron per molecule. Neither chromate titration nor chromate colorimetry is sensitive enough for organic matter measurement in most soil solutions. Formation of insoluble intermediate Mn oxides preclude use of permanganate in stepwise colorimetry. In short, Mn(III) offers unique advantages as an oxidizing agent.