History of all developments in the chemistry of soil Organic matter.

Click to edit Master subtitle style

Rajan Bhatt L-2010-A-97-D

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History of organic structure upto nowJust a fast look

Dark coloured amorphous precipitate on acidification of the alkaline extract by Achard (1786) who extracted peat with dilute base . Desaussure (1804) describe it as humus Sprengel (1826) observed that in soils rich in Ca and Mg, humus was present in bound form. Berzelius (1839) separated two light-yellowish substances from mineral waters and mud rich in iron oxides, which he named as crenic and apo-crenic acids Herman (1842) found that humus was complex in nature.

Schnitzer (2001), Stevenson.F.J. (1981) Humus Chemistry-Genesis, Composition , Reactions. Chapter 2 and William4/28/12 (1915)

Contd.

Malaguti (1858) that humic acid and humin are formed when sugars is boiled with either HCL or H2SO4. Conrad and Guthzeit (1885) found the dextrose and laevulose, further dextrose formed humic acid anf laevulose humin on boiling suger with HCL acid. Hopper-style (1889) added phlobaphrene dissolved in alcohol from acid ppt. of humic acid group.
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Contd.

Irvin and Dobson (1907) observed a structural difference exists between the arttificial and natural acids. Baumann (1909) freshly ppt. HA possess colliodal properties especially the power of forming the adsorption compounds. Schreiner and Shorey (1910)giving grouping for various compounds that tyhey isolate from Organic matter. Stevenson (1994), De Saussure (1804) introduced the term humus to describe the dark-coloured material extracted from soils by dilute alkali.

Schnitzer (2001), Stevenson.F.J. (1981) Humus ChemistryGenesis, Composition , Reactions. Chapter 2 and William (1915)
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Difficulties in extraction with alkaline solutions (Stevenson, 1994) Silica dissolved in minerals may
contaminate the SOM extract. Protoplasmic and structural components of fresh organic tissues are dissolved and these mix with SOM extract. Auto-oxidation of Organic material in the air Condensation between amino acids and keton groups
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During Late 1940

Wet

chemistry was done using Beakers, Flasks, Test tubes, Instruments 1. pH meter (Powered by batteries) The Danish biochemist Soren Sorensen then invented the pH scale in 1909 2. Colorimetryis "thescienceandtechnologyused to quantify and describe physically 4/28/12 the humancolor perception."It is

Differences b/t colorimeter and spectrophotometer

COLORIMETER SPECTROPHOTOM 1. Consists of sensor plus 1. Consists of ETER processor data or computer with software. sensor and simple data 1. Isolates a narrow band of wavelengths. processor. 2. Is a more complex
1.

2.

Isolates a broad band of wavelengths. Is generally rugged and a

instrument colorimeter.

than

1.

Works well for color formulation, measurement of metamerism, (which relates to two colors and their relation under different light sources). 4/28/12

Ph meter Colorimeter

The Danish biochemist Soren Sorensen then invented the pH scale in

(1) Wavelength selection, (2) Printer button, (3) Concentration factor adjustment, (4) UV mode 4/28/12

Disadvantages of pH meter:
1. AllpH metersneed to be calibrated at some frequency. 2. The probe has to be cleaned well after every Click to edit Master subtitle style use to make sure you get a reliable readingnext time pH readings are affected by temperature. 3. By pH meter we are able to now that which field had higher organic matter in it but we dont know the individual composition of organic matter.

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Disadvantages of colorimeter

Igloo writes: You need to carry out an error analysis using the titration methods and compare your results with those obtained from carrying out the same analysis with the colorimetry as it is not accurate. Complexity of the machine that can be intimidating or confusing to learn to use. The interpretation of data is more complex than with spectrophotometers. Colorimeters have poor inter-instrument agreement; two colorimeters may give different readings for the same sample. The lack of standard observer angle, geometry, or illuminant can lead to varying results among operators or readings.
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SPECTRUM OF ELECTROMAGNETIC RADIATION ORGANISED BY INCREASING WAVELENGTH FROM LEFT TO RIGHT 1 0 10 2

UV

X-ray

10 3 1 Visible m

10 4
IR

Wavelength in nm 10 10 10 10 10 5 Microwave 8 91 6 71c m m

Radio

Change in electronic states X-ray Absorption UV/Vis absorption

Change in mol. Rot Change in rotational states and Vib states of IR spectroscopy Electron spin ESR Nuclear spin NMR

During early 1950s

Ultraviolet/visible spectrophotometer: It is absorption & reflection spectroscopy in UV (200-400nm)/ visible (400-800nm) spectral region, thus uses light in the visible and adjacent ranges as absorption in visible range directly affects the colour of chemical which is because of chromophores (=bond). This is complementary to fluorescence spectroscopy.
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Recording ultraviolet spectrophotometer

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UV

spectroscopy has limited use in analyzing mixtures, due to the addition of absorbance it requires special equipment (a UV light source and UV-transparent sample holders) and it is not selective for compounds if they absorb light spectra at the same wavelength
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Limitation of Ultraviolet/visible spectrophotometer

Mid 1950s
Infrared

spectro-photometer:

Thespectroscopythat deals with theinfraredregion of theelectromagnetic spectrum, that is light with a longer wavelength thanvisible light. It covers a range of techniques, mostly based onabsorption spectroscopy. As with all spectroscopictechniques,itcanbeusedto identifyandstudychemicals.
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IR spectroscopy provide information on the presence of specific functional group within a molecule on the basis of absorption band (its position and intensity) related to that functional group. Presence of that particular band depicts the presence of that particular functional group in molecule in other case it shows its absence.

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Limitation of Infrared spectrophotometer

Infrared spectrophotometer will not allowed us to separate complex mixtures of humic acid and Fulvic acid oxidation products along with organic soil extracts . but with the discovery of Gas chromatograph this problem is solved as it is able to perform the above separation. One major drawback of IR spectroscopy is the interference (patterns which may be observed when two light waves are superposed) caused by adsorbed moisture, which produces the band in the 3300 to 3000 and 1720 to 1500 cm-1

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Separates

During 1960s-Gas Chromatograph-

mixtures, especially very volatile liquids

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Gas chromatograph

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Limitation of Gas chromatograph

Gas

chromatography can only be used in cases where the substances can be vaporized without decomposing and where they can be vaporized at a reasonable temperature (i.e. not so hot that it destroys the column packing). The samples must be thermally stable to prevent degradation when heated. 4/28/12 Cannot be used to prepare samples

During Mid 1960s

Mass spectro-metery:
is

an analytical technique that measures themass-to-charge ratioof charged particles.It is used for determining masses of particles, for determining the elemental composition of a sample.

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Mass spectrometers

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Mass Spectrometer
All Instruments Have:
1. 2. 3. 4. 5.

Sample Inlet Ion Source Mass Analyzer Detector Data System

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Disadvantage of Mass spectrometery

1. Doesn't directly give Master subtitle style Click to edit structural information. 2. Needs pure compounds 3. Difficult with non-volatile compounds

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During Mid 1960s

Electron spin resonance (ESR) is a sophisticated spectroscopic technique that detects free radicals or inorganic complexes in chemical and biological systems (have one or more unpaired electrons). It also throws light on the symmetry and coordination of paramagnetic ions viz. Fe3+, Cu2+ and Mn2+ bound to HA and FA. Most stable molecules only have paired electrons without any free radicals, so ESR Spectroscopy is used less often than NMR
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ESR Spectrometers

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Disadvantages of ESR (Electron spin resonance )

Loss of resolution of the signals which is determined by the overlap of component lines to such an extent that information is lost. stable molecules have all their electrons paired, then there ESR technique is less widely used than NMR as here we focus on movement of nuclei instead of electrons. However, this limitation toparamagneticspecies also means that the ESR technique is one of great specificity, since ordinary chemical 4/28/12

Early 1980s-13C- Nuclear Magnetic Resonance (NMR) Spectrometer

NMR

is the most powerful tool available for organic structure determination. It is used to study a wide variety of nuclei:

1H 13C 15N 19F 31P

Chapter 13

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28

13C- Nuclear Magnetic Resonance (NMR) Spectrometer. Nuclear magnetic resonance (NMR) is a property that magnetic nuclei have in a magnetic field and applied electromagnetic (EM) pulse or pulses, which cause the nuclei to absorb energy from the EM pulse and radiate this energy back out. The energy radiated back out is at a specificresonancefrequency which depends on the strength of the 4/28/12 magnetic field.

Nuclear Spin
A

nucleus with an odd atomic number or an odd mass number has a nuclear spin. The spinning charged nucleus generates a magnetic field.

=>
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. Importance on 13 -C NMR : First time It shows that aliphatic C in HA and FA is equally important as that of aromatic C and the Older theories that HA consist of completely with aromatic were no longer.
(Aromaticity seldom exceeds than 55% and the aliphaticity of SOM often greater than aromaticity, Schnitzer and Preston, 1986)
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The NMR Spectrometer

=>
Chapter 13 4/28/12 32

Limitation of 13-C NMR

1.

2.

Direct analysis of OM in whole soils is not possible. We have to go for tedious processes of Extraction and purification of the Organic Matter for its analysis. 13C NMR detects only the13 Cisotopeof carbon, whosenatural abundanceis only 1.1%, but the main carbon isotope,12 C, is not detectable by NMR since it has zero 4/28/12 netspin.

During mid 1980s

Importance of Pyrolysis- Field Ionization Mass Spectrometery (Py- FIMS) Till now we are not possible to Direct analysis of OM in whole soils without Extraction and purification as even in 13c NMR also we have to go first for Extraction and Click to procedures but with this purification tedicious edit Master subtitle style we could easily analyze OM without extraction and purification as The pyrolysis enables non-volatile substances to be analysed gas chromatographically.

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Py- FIMS- Uses

structural

studies of the humic acid (2D structure) which further to 3-D structural model Kruse et al., (2010) observed that PyFIMS spectra reflected differences in DOM composition related to land use and sampling depth. Land use effects were expressed in higher abundances of carbohydrates and peptides at the arable than at the fallow site. The 4/28/12 relative proportions of carbohydrates

Schnitzer 4/28/12

and

Cultivation causes increased polymerization and cross linkages of major soil components leading to Schnitzer and formation of large molecules with 4/28/12

 GC-MS

is a method that combines the features ofgas chromatographyandmass spectrometryto identify different substances within a test sample. Applications of GC-MS includedrugdetection,and identification of unknown samples. The use of a mass spectrometer as the detector in gas chromatography was developed during the 1950s by Roland Gohlke andFred McLafferty. 4/28/12

GCMS (Gas chromatography Mass spectrometery)

Mass spectrometer attached` with Gas chromatograph (GCMS) which allows us not only to separate but also to identify these separates (GCMS)

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GCMS--- Uses
Identification and quantitation of volatile and semivolatile organic compounds in complex mixtures Determination of molecular weights and elemental compositions of unknown organic compounds in complex mixtures Structural determination of unknown organic compounds in complex mixtures both by matching their spectra with reference spectra and by a prior spectral interpretation

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GCMS--- Limitations
Only compounds with vapor pressures exceeding about 1010 torr (1 torr=1 0.001316 atmosphere) can be analyzed by GC-MS. Qualitative accuracy is restricted. Depending on the dilution factor and ionization method, an extract with 0.1 to 100 ng of each component may be needed in order to inject a sufficient amount.

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Electron Microscopy
Size

and shape of colloidal particles such as HS and their morphological conformation can be directly obsereved by use of electrons.

SEM

(Scanning electron microscopy) TEM (Transmission electron microscopy)

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TEM

Transmission electron microscopy(TEM) is amicroscopytechnique whereby a beam ofelectrons is transmitted through an ultra thin specimen, interacting with the specimen as it passes through. An image is formed from the interaction of the electrons transmitted through the specimen; the image is magnified andfocusedonto an imaging device, such as afluorescentscreen, on a layer ofphotographic film, or to be detected by a sensor such as aCCD camera. TEM can produce very high-resolution images of a sample surface, revealing details not more than 50nmin size Electron beam is used to magnify the surface properties. Orlov et al., 1975 see spherical aprticles with 6-10 nm dia and M.WT of > 20,000 Da. Wershaw and aiken (1985) observed a HA particle with an Dia of 2nm.
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TEM

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SEM

Ascanning electron microscope(SEM) is a type ofelectron microscope that images a sample by scanning it with a high-energy beam ofelectronsin araster scan pattern. The electrons interact with the atoms that make up the sample producing signals that contain information about the sample's surfacetopography, composition, and other properties such aselectrical conductivity. SEM can produce very high-resolution images of a sample surface, revealing details less than 1nmin size Different from TEM as here images are formed from the secondary elctrons emitted from sample surface when electron beam bambarred over the surface. Orlov et al., (1975) observed featureless sheets of condensed material. Under SEM at Ph 2-7, FA and HA exhibited shape of elongated linear fibers that tended to become thinner on increasing the Ph and bundles of fibers tended to become predominant at Ph 6 and to a fine network at Ph 7. Ghosh and Schnitzer (1982) observed increase in fiber thickness and decrease in particle orientation with increasing conc. Of neutral 4/28/12 salts in soilution of HA and FA

SEM

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Drawbacks
Sample

preparation procedures is more complex here than in other techniques. Conformation of HS macromolecules is strongly affected by conditions of initial sample such as Ph, sample conc., ionic strength and solvent characteristics which further strongly effect charge, hydrogen-bonding , disperstion and aggregation of HS 4/28/12 particles

Drawback
As

a result , the size and shapes of HS observed by electron microscopy are unlikely to represent the size and shape of HS that actually present in the soils.

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Conclusion
Recently there is no universally accepted method for the extraction, fractionation and purification of HS. Tremendious efforts are being made by Intrernational Humic substance Society ( IHSS) Click to edit Master subtitle of standarized, to encourage the adoption style unique, comprehensive procedure for isolation of HS from the soils, sdeiments and water. (IHSS was founded in Denver, Colorado, USA, on September 11, 1981 to bring together scientists in the coal, soil, and water sciences with interests in humic substances. IHSS has a membership of nearly 900 scientists).
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Refrences
Arshad, M. A., Ripmeester, J. A. and Schnitzer, M. 1988. Attempts to improve solid state 13C NMR spectra of whole mineral soils. Can. J. Soil Sci. 68: 593602 Schulten, H.-R., Monreal, C. M. and Schnitzer, M. 1995. Effect of long-term cultivation on the chemical structure of soil organic matter. Naturwissenschaften 82: 42 44.

Kruse et al (2010) Pyrolysis-field ionization mass spectrometry and nitrogen K-edge XANES spectroscopy applied to bulk soil leachates--a case study. Sci Total Environ.2010 Sep 15;408(20):4910-5. Epub 2010 Jul 24. Schnitzer, M and Schulten, H.R. 1992. The analysis of soil orgfanic matter by pyrolysisfield ionization mass formation of humic William B.B (1915) The spectrometry. Soil sci. Soc. Am. J: 1811-1817 bodeis from Organic substances. Schnitzer, M and Schulten, H.R. 1995. 4/28/12

Thanks
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An historical account of extraction procedures for SOM in soils and its subsequent fractionation

The first attempt to separate OM from soils appears to have been made by Achard (1786), who extracted peat with dilute base and obtained a dark coloured amorphous precipitate on acidification of the alkaline extract. Achard observed that larger amounts of OM could be extracted from the lower, more decomposed layers of the peat than from the upper, less decomposed layers. According to Stevenson (1994), De Saussure (1804) is credited with introducing the term humus (Latin for earth) to describe the dark-coloured material extracted from soils by dilute alkali.

Scnitzer. M (2001)

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Cont..
The first systematic chemical study on humus was carried out by Sprengel (1826), who observed that in soils rich in Ca and Mg, humus was present in bound form, and this soil was highly fertile. But, in soils low in bases, humus occurred in the free form, which rendered the soil acid and unproductive. It was Berzelius (1839), the outstanding Swedish chemist, who separated two light-yellowish substances from mineral waters and mud rich in iron oxides, which he named crenic and apocrenic acids. These acids were soluble in acid and base and are today referred to as fulvic acid. Scnitzer. M (2001)
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Contd.
Berzelius found that these acids could form watersoluble complexes with a number of metals. Berzelius distinguished the following humus (= total SOM) components: (1) humic acid (HA), soluble in alkali but coagulated by acid; (2) the inert humin, which could not be extracted by alkali; and (3) the alkali- and acid-soluble crenic and apocrenic acids. A similar classification of humus (total SOM) according to colour and solubility in water and alkaline solutions was proposed by Mulder (1861). Scnitzer. M (2001)
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Contd..
Mulder (1861) divided total SOM into the following fractions: (1) insoluble in alkali ulmin and humin; (2) soluble in alkali ulmic acid (brown) and HA (black); (3) soluble in water crenic (dark yellow) and apocrenic (light yellow) acids. The early workers believed that ulmin, humin, ulmic acid, HA, crenic and apocrenic acids were chemically pure compounds. They determined their elementary composition and empirical formulae. The fractionation procedure currently in use does not differ significantly from that proposed by the early workers except that it has been simplified. Scnitzer. M (2001)
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Hermann concluded that when humic acids were oxidised to crenic and apocrenic acids, still more N was absorbed. Later, HopperSeyler derived Hymatomelanic acids by extracting humic acid with alcohol. From 19001940, Oden lassified humic substances into humus coal, humic acid, hymatomelanic acid and fulvic acid here humus coal represent the humin and ulmin of Berzelius and Mulder and is insoluble in water, alcohol and alkali.
Stevenson.F.J. (1981) Humus Chemistry-Genesis, Composition , Reactions. Chapter 2,
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During 1908, Schreiner and shorey started studies on identifiable organic chemicals in soils and upto next 30 years they identifies around 40 compounds viz. organic acids, fats, sterols etc. Shmook (1930) reviewed imp. Aspects of humus chemistry and its formation by soil organisms. He also showed the presence of COOH group in humic acids. Further he concluede that HA has an organic Ncontaining compouns and aromatic rings derived from the lignin. Stevenson.F.J. (1981)
Humus Chemistry-Genesis, Composition , Reactions. 4/28/12

Hopper-Seyler derived Hymatomelanic acids by extracting humic acid with alcohol. Oden classified humic substances into humus coal, humic acid, hymatomelanic acid and fulvic acid.
Schnitzer (2001), Stevenson.F.J. (1981) Humus Chemistry-Genesis, Composition , Reactions. Chapter 2 and William (1915)
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Waksman states that Humus comprised of fats, waxes, Cellulose, hemicellulose, liganoprotein with protein derived from microorganisms and gave proximate analysis technique. Springer on the basis of optical properties and behavior toward electrolytes, divide HA into Braunhuminsaure and Grauhuminsaure. Waksman isolated separate fraction from fulvic acid and but later recommend the use of Humus instead of Humic acid, Fulvic acid and Humin. Schnitzer (2001), 4/28/12 Stevenson.F.J. (1981)

Mulder

(1861) classify humus into HA, Humin and the alkali- and acidsoluble crenic and apocrenic acids. Hermann oxidised humic acids to crenic and apocrenic acids. HopperSeyler derived Hymatomelanic acids by extracting humic acid with alcohol. Oden classified humic substances into humus coal, humic acid, hymatomelanic acid and fulvic 4/28/12 acid.

Schreiner (1908) identifies around 40 compounds viz. organic acids, fats, sterols etc. Shmook (1930)showed the presence of COOH group in humic acids. Millard, states that HS does-not involved directly to produce sugar from carbohydrates and amino acid from proteins
Stevenson.F.J. (1981) Humus Chemistry-Genesis, Composition , Reactions. Chapter 2,
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Trusov states that humification process has following order 1.Hydrolytic decomposition of plant. 2. Oxidation of later by enzymes klead to hydroxy quinone 3. condensation of quinone in dark coloured products (HS). Williams gave two stage of humification 1. decompostion of original plant residues to simpler compounds 2. synthesis of substances more complex in nature. Fuchs, Hobson, Page and Waksman and co-workers advances the theory that Lignin was the precursor of Humic Acids. Stevenson.F.J. (1981) Humus Chemistry-Genesis, Composition , Reactions. Chapter 2,
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Waksman states that Humus comprised of fats, waxes, Cellulose, hemicellulose, liganoprotein with protein derived from microorganisms and gave proximate analysis technique. Springer on the basis of optical properties and behavior toward electrolytes, divide HA into Braunhuminsaure and Grauhuminsaure. Waksman isolated separate fraction from fulvic acid and but later recommend the use of Humus instead of Humic acid, Fulvic acid and Humin. 4/28/12

Then Trusovs concept of humification (reviewed by Kononova) is quite close to present day concept as he focused that humification process has following order 1.Hydrolytic decomposition of plant remains lead to synthesis of simple substance of aromatic nature. 2. Oxidation of later by enzymes klead to hydroxy quinone 3. condensation of quinone in dark coloured products (HS). Later Williams gave two stage of humification 1. decompostion of original plant residues to simpler compounds 2. synthesis of substances more complex in nature. Fuchs, Hobson, Page and Waksman and co-workers advances the theory that Lignin was the precursor of Humic Acids. 4/28/12

Afterward Waksman regarded that Humus comprised of fats, waxes, Cellulose, hemicellulose, liganoprotein with protein derived from microorganisms and gave proximate analysis technique. Springer on the basis of optical properties and behavior toward electrolytes, divide HA into Braunhuminsaure and Grauhumin saure. Waksman isolated separate fraction from fulvic acid and but later recommend the use of Humus instead of Humic acid, Fulvic acid and Humin. 4/28/12

Modern day concept

Humus can be divided into Nonhumic Substances and Humic substances. However two groups are not easily separated because some non-humic substances viz. carbohydrates may bound covalently to the humic matter. Humus probably contain most, if not all, of the biochemical compounds synthesized by living organisms.
Stevenson.F.J. (1981) Humus
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Chemistry-Genesis,