Physico-chemical Attributes of Humic Acid Extracted from ... PAPERS/JTAS Vol. 19 (2&3) Dec....

Pertanika J. Trop. Agric. Sci. 19(2/3): 189-196 (1996)ISSN: 0126-6128

i Penerbit Universiti Pertanian Malaysia

Physico-chemical Attributes of Humic Acid Extracted fromTropical Pect

M.H.A HUSNI, SHANTI DEVI, ABD.RAHMAN MANAS1 and K.B. SIVADepartment of Soil Science

Faculty of AgricultureUniversiti Pertanian Malaysia

43400 Serdang, Selangor, Malaysia

Department of ChemistryFaculty of Science and Environmental Studies

Universiti Pertanian Malaysia43400 Serdang, Selangor, Malaysia

Keywords: humic acid, tropical peat, infrared spectra

ABSTRAKKajian asas mengenai asid humik pada gambut tropika adalah terhad. Satu kajian mengenai ciri-ciri asid humik

yang diekstrak dari gambut tropika telah dijalankan dengan menganalisis elemen dan kumpulan berfungsi, spektruminframerah dan ujian terus pada asid humik dengan menggunakan mikroskop elektron imbasan. Berdasarkan kepadaanalisis elemen, jumlah Cyang didapati adalah 40.26 - 53.26%, H 4.84 - 6.36%, 0 38.39 - 51.62%, JV L51 -2.64%, dan P 0.02 - 0.06%. Kajian dalam kumpulan berfungsi menunjukkan kumpulan karboksil yang wujuddalam asid humik di gambut tropika adalah dua kali ganda lebih banyak daripada kumpulan hidroksil fenolik.Daripada spektrum inframerah, asid humik didapati mempunyai ciri-ciri aromatik. Analisis inframerah jugamenunjukkan spektrum yang sama dan selaras tanpa mengira perbezaan tempat asalnya, kompaun humik yangdiekstrak mempunyai struktur dan komposisi kumpulan berfungsi yang sama. Pemeriksaan menerusi mikroskopelektron pada asid humik kering menunjukkan bentukyang berblok dan separa berliku dengan saizyang berbeza.Semua asid humik yang diekstrak menunjukkan bentuk yang sama.

ABSTRACTBasic research conducted on humic acid (HA) in tropical soils is limited. A study was conducted to characterize HAextracted from tropical peat by elemental and functional group analyses, infrared (IR) spectra and by directexamination on HA using scanning electron microscopy (SEM). From the elemental analysis it was found that Cranged from 40.26 - 53.26%, H 4.84 - 6.36%, 0 38.39 - 51.62%, N 1.51 - 2.64%, and P 0.02 - 0.06%. Studies offunctional groups indicated that the carboxyl group present in HA of tropical peat was about twice as high as that ofphenolic hydroxy I groups. From IR spectra, the HA had an aromatic character. Results of IR analysis also exhibitedconsistently similar spectra, demonstrating that regardless of place of origin, the extracted humic compound had asimilar structure and was composed of the same functional groups. Electron microscopic examination of dried HAshowed a sub-angular blocky shape which varied in size. All extracted HA exhibited similar form.

INTRODUCTION

Humic acid (HA), defined as the fractionthat is soluble in dilute base and coagulatedupon acidification of the alkaline extract, isthe main component of humic fraction inboth temperate and tropical peat (Nor-hayati 1989; Garcia et al. 1991). HA is

comprised of highly reactive polymericcompounds of varying molecular weight,containing C, H, O, and N, and itsreactivity is influenced by the nature andamount of oxygen-containing functionalgroups (Stevenson and Goh 1972).

According to Cheshire et al. (1967),

M.H.A HUSNI, SHANTI DEVI, ABD.RAHMAN MANAS AND K.B. SIVA

peat humic acids have 52% C, 5.1% H,2.2% N and 40.3% O content. A study byVisser (1987) on humic acids extractedfrom decomposing plant and peat coresamples showed content of 34-55% C, 5-11% H and 40-55% O, different from thevalues of mineral soils 55-60% C, 3-5% H,2-6% N and 30-40% O (Kononova 1961).HA extracted from Sphagnum and Carexpeat showed 49.7 and 55.4% C, 6.0 and5.0% H, 42.0 and 35.5% O, 2.3 and 3.5% Ncontent, respectively (Garcia et al. 1991).

IR spectra of HA extracted fromtemperate peat showed aromatic characterand great similarity among different sam-ples of HA (Visser 1987; Garcia et al. 1991).Studies by Norhayati (1989) also indicatedthat the HA extracted from tropical peatexhibited very similar IR spectra.

SEM provides a non-destructive meth-od for direct recording of the shape of tinyhumic particles (Bohr and Hughes 1971;Orlov and Glebova 1972). Ghosh andSchnitzer (1982) found under SEM, humicacids to consist of small discrete sphenoid,flattened aggregates, linear chain assem-blies of these aggregates and flattenedfilaments.

There is a dearth of information on thestudy of elemental composition and reac-tivity of HA, and its structure under SEM.This study was therefore carried out tocharacterize the HA extracted from tropi-cal peat by elemental and functional groupanalyses, its IR spectra and its structure bySEM.

MATERIALS AND METHODS

Fourteen samples from major peat areas ofMalaysia were used in this experiment,twelve from the west coast of PeninsularMalaysia and two from Sarawak (peatsamples 5 and 6). Percentage of organicmatter was determined as loss on ignitionfrom sample maintained at 500°C for 4 h(Storer 1984).

Humic acid was extracted with 0.2 MNaOH using a soil to extractant ratio of1:10. The mixture was shaken continuouslyon a mechanical shaker for 24 h and thesoluble organic matter was separated fromthe peat soil by centrifugation at 10,000rpm for 15 min. The residue was washedwith distilled water in a ratio of 1:5 andwashing water separated from the residueby centrifugation. The combined super-natants were acidified with concentratedH2SO4 to pH 1.0 and allowed to stand for24 h. The coagulated humic acid (HA)precipitate was purified by re-precipitationand re-dissolution using H 2 SO 4 andNaOH, respectively. The alkaline extractwas then shaken for 48 h with excess HC1-HF mixture (0.5%, v/v). The acid mixturewas removed by centrifugation and theresidue thoroughly washed with distilledwater until the filtrate became free ofchloride. The humic acid was then dia-lyzed against distilled water until there wasno colour change in the outer solution(distilled water) and finally frozen anddried at ambient temperature (Schnitzer1982; Norhayati and Verloo 1984).

Carbon, H and N were determined bydry combustion using a CHN analyser(VARIO-EL), O was calculated by differ-ence and P by dry ashing at 500°C anddetermined colorimetrically at 882 nm(Murphy and Riley 1962). Carboxylgroups were measured by Ca(OAc)2 meth-od (Schnitzer and Gupta 1965) and thephenolic hydroxyl groups by the colori-metric method using Folin Cicocalteu'sreagent (Tsutsuki and Kuwatsuka 1978).These determinations were represented asthe average result of two duplicate samples.

IR spectra were recorded on a Beck-man Fourier transfer IR spectrophotometerusing the KBr pellet technique. Prior to theanalyses, the HA with the KBr was storedfor at least 48 h in a desiccator to preventabsorption of atmospheric moisture.

190 PERTANIKA J. TROP. AGRIC. SCI. VOL. 19 NO. 2/3, 1996

PHYSICO-CHEMICAL ATTRIBUTES OF HUMIC ACID EXTRACTED FROM TROPICAL PECT

For SEM examination, the dried HAobtained directly after extraction wasattached to Al-stubs with the aid ofcolloidal silver paint. To make the speci-mens surface conducting, they were coatedwith a layer of gold-palladium. Theprepared specimen were examined on aSEM (JOEL 35C) operated at 25kV.

RESULTS AND DISCUSSION

The organic matter contents per 100 g peatof the 14 peat samples studied ranged from44 - 97% (Table 1). Percentage of HAobtained ranged from 8.30 - 39.52/100 gpeat.

The data on the elemental compositionof HA in terms of percentage weight arepresented in Table 2. Carbon contentranged from 40.26 - 53.26%, H 4.84 -6.3%, O 38.39 - 51.62%, N 1.51 - 2.64%and P 0.02 - 0.06% with means of 49.34%C, 5.64% H, 43.05% O, 1.99% N and0.03% P. Comparing these with theelementary composition of HA reported

by Visser (1987) and Garcia et aL (1991)indicated that our results fell within thereported range for temperate peat.

The analysis of the major oxygen-containing functional groups is shown inTable 3. The carboxyl group of HAextracted from tropical peat ranges from2.50 - 3.10 meq/g of HA while phenolichydroxyl groups range from 0.99 - 1.75meq/g of HA. Mean values of functionalgroups showed 2.84 meq COOH and 1.36meq phenolic hydroxyl/g of HA. Thecarboxyl groups present in HA are morethan twice as numerous as in the phenolichydroxyl groups, implying that the pH-dependent charge in peat is controlled bycarboxyl groups.

IR spectra and the absorption bands ofHA extracted from peat are shown in Fig. 1and Table 4, respectively. The broadabsorption band at 3550-3200 cm"1 regioncan be attributed to H-bonded OH groups,while peaks between 2920-2860 cm"1 aredue to aliphatic C-H stretch. According to

TABLE 1Classification and yield of HA from peat

PeatSample

No.

123456789

1011121314

1 Classification

AAABABAABAABBA

Depth (cm)2

(%)

0-150-150-15

15-300-15

15-300-150-150-150-20

20-400-15

20-400-15

O.M.3

(%)

4482868890949797769497899796

HA

21.2326.8818.3025.7823.788.40

35.6720.6823.4536.5212.3819.828.30

26.92

1 A Typic Tropohemists, B Typic Tropofibrists2 Percentage of organic matter3 % Humic acid (HA) per 100 g peat -

PERTANIKA J. TROP. AGRIC. SCI. VOL. 19 NO. 2/3, 1996 191

M.H.A HUSNI, SHANTI DEVI, ABD.RAHMAN MANAS AND KB. SIVA

TABLE 2Elemental analysis of HA extracted from tropical peat

PeatSample

No.

123456789

1011121314

Range

Mean

C

51.9352.0145.7551.2553.3944.3951.0447.8049.7650.5249.0750.0053.5640.26

40.26-53.56

49.34

H

5.145.626.365.235.596.235.225.774.845.145.785.675.916.32

4.84-6.36

5.64

Elements (%)

O

40.5239,8845.2641.3338.3447.7241.9844.6843.8842.1443.4842.1638.3851.60

38.39-51.62

43.05

N

2.372.452.592.162.641.631.741.731.512.181.651.711.641.84

1.51-2.64

1.99

P

0.060.040.050.040.050.030.020.020.020.030.020.020.020.02

0.02-0,06

0.03

TABLE 3Analysis of oxygen-containing functional groups of HA extracted

from tropical peat

Peat Functional groups (meq/g of HA)SampleNo. COOH Phenol OH

1.361.071.121.390.991.561.221.441.401.751.431.541.591.22

Range 2.50-3.10 0.99-1.75

Mean 2.84 1.36


123456789

1011121314

2.902.752.952.752.803.102.502.802.903.003.002.852.952.50


TABLE 4Absorption bands of infrared spectra of HA from tropical peat

Bands(cm'1)*

PeatSampleNo.

1*23456789101112131314

3550-3200

s +

ssssssssssssss

2920-2860

mmmmmmmmmmmmmmm

1725-1705

- Relative

mmwmmmwmmwmmmmm

1690-1600

sssssssssssssss

1550-1500

aaaaaawwwwwwwww

1470-1430

wwwwwwwwwwwwwww

1280-1200

wwwwwwwwwwwwwww

* 3550-3200 (hydrogen-bonded OH); 2920-2860 (aliphatic C-H stretch); 1725-1705 (C = Oof COOH and ketones); 1690-1600 (C = O stretch of quinones, C = O and/or C = O of Hconjugated ketones); 1550-1500 (aromatic ring stretch, P- and O- distributed); 1470-1430(aromatic ring stretch, O-distributed); 1280-1200 (C-O stretch, and deformation of COOH,C-O stretch of aryl ethers)+ (s) strong; (m) medium; (w) weak; (a) absent# Sample number for HA extracted from respective peat soil

Bellamy (1969), bands in the region of3000-2500 cm"1 were produced by mostcarboxylic acids and arose from vibration ofintermolecular OH...O band. Therefore,Schnitzer and Griffith (1975) indicatedthat the O-H stretch (<2800 cm"1) was atest for the occurrence of H-bonded COOHgroups. However, in this study only amedium peak occurred at this regionsimilar to the results of Norhayati (1989).

Norhayati (1989) showed that higheramounts of carboxyl groups were present inHA molecules. Similar results were ob-tained in this study, where most of the HAspectrum exhibited a medium peak in the1725-1705 cm"1 region (band for carbonyl

group) with strong peaks in the 1690-1600cm"1 region (band for carbonyl group).

The spectra also showed weak bands at1550-1500 cm"1 and 1470-1430 cm"1.Similar results were obtained by Garcia etal. (1991), where the above bands meantfor HA extracted with alkali disappearedfor HA extracted with pyrophosphatereagents in both Sphagnum and Carexpeat. This indicated that HA extractedwith alkali extractant exhibited strongeraromatic characteristics than that usingpyrophosphate reagent.

A weak band obtained for all samplesin the 1289-1200 cm"1 region is attributedto OH, from COOH deformation. Other



4000 3000 2000 1600 1200 800 400

cm"Fig. 1. Infrared spectra of HA extracted from

tropical peat

bands, such as 1440, 1125 and 1034-1080cm"1 were also observed. However, all thesebands were of weak intensity, and wereabsent in some samples {Fig. 1). Bandsoccurring at 1440 cm'1 indicated the pre-sence of the carboxyl anion (SchnitzerandGriffin 1975); 1125 and 1034-1080 cm1

indicated aromatic ring P- and O- disub-stituted, C-O-O stretch of aryl ethers andC-O stretch, aromatic ether, hydratedpolyols and carbohydrates, respectively(Garcia et aL 1991). Thus, this studydemonstrates that HA of tropical peat hasa greater amount of carboxyl groups.

The IR spectra of the HA extractedfrom peat of diverse origin displayed similarbands and indicated the presence of similarstructures, differing mainly in the numberof functional groups. The results corre-sponded with those of Norhayati (1989).

The shape and particle arrangement ofHA under SEM are shown in Plate 1. TheHA materials are sub-angular blocky, varyin size, and are irregular in shape {Plate la).Plate lb shows the plate-like structure of HAsurface upon higher magnification (1 |im).

Plate 1. Scanning electron micrographs of shape andparticle size humic acid; a) top: sub-angular blocky whichvary in size and shape, b) bottom: plate like structure of

HA surface.



CONCLUSION

The elemental analysis of HA showed Cranged from 40.26 - 53.26%, H 4.84 - 6.36%,O 38.39 - 51.62%, N 1.51 - 2.64% and P0.02 - 0.06%. Studies of the functionalgroups indicated that carboxyl groupspresent in HA of tropical peat are abouttwo times more numerous than phenolichydroxyl groups. From IR spectra, the HAobtained is of aromatic character. The IRanalyses also exhibited consistently similarspectra, demonstrating the possibility thatregardless of sample site, the extracted HAhas a similar structure and is composed ofthe same functional groups. SEM examina-tion of dried HA showed sub-angularblocky shape of varying size was similarfor all the HA extracted. These resultssupport those obtained by Norhayati(1989) for Malaysian peat, and demon-strate the similarity between tropical andtemperate peats regardless of plant originor climate.

ACKNOWLEDGEMENT

The authors wish to thank the NationalCouncil for Scientific Research and Devel-opment, Malaysia, Program 1 -07-05-047for financial support, and Mr. Mohd FuziMohd. Sharif of Soil Fertility Section for histechnical assistance.

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(Received 1 August 1996)(Accepted 31 January 1997)


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