Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 116 (2013) 642–645

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Spectrochimica Acta Part A: Molecular andBiomolecular Spectroscopy

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Short Communication

Fourier transform-infrared spectroscopy and Gas chromatography–massspectroscopy: Reliable techniques for analysis of Parthenium mediatedvermicompost

1386-1425/$ - see front matter � 2013 Elsevier B.V. All rights reserved.http://dx.doi.org/10.1016/j.saa.2013.08.012

⇑ Corresponding author. Tel./fax: +91 4222611146.E-mail address: rajeshwarishivaraj@gmail.com (S. Rajeshwari).

P. Rajiv a, Sivaraj Rajeshwari a,⇑, Rajendran Venckatesh b

a Department of Biotechnology, School of Life Sciences, Karpagam University, Eachanari post, Coimbatore 641 021, Tamil Nadu, Indiab Department of Chemistry, Government Arts College, Udumalpet 642 126, Tamil Nadu, India

h i g h l i g h t s

� Parthenium hysterophorus L. is anoxious weed plant and toxic tohumans and animals.� Biomass of Parthenium was used as a

raw material for production ofvermicompost.� FT-IR spectroscopy of vermicompost

showed reduction in toxic compound.� GC–MS reveals that analysis of

compounds in vermicompost andcompost.

g r a p h i c a l a b s t r a c t

FT-IR spectra of initial feed mixture and vermicompost obtained from 60% cow dung + 40% Parthenium.

a r t i c l e i n f o

Article history:Received 19 April 2013Received in revised form 28 July 2013Accepted 2 August 2013Available online 13 August 2013

Keywords:Parthenium hysterophorusSesquiterpene lactoneVermicompostFT-IRGC–MSEudrilus eugeniae

a b s t r a c t

Fourier transform infrared spectroscopy (FT-IR) and Gas chromatography–mass spectroscopy have beencarried out to investigate the chemical composition of Parthenium mediated vermicompost. Four differentconcentrations of Parthenium and cow dung mixtures were vermicomposted using the earthworms(Eudrilus eugeniae). FT-IR spectra reveal the absence of Parthenin toxin (sesquiterpene lactone) and phe-nols in vermicompost which was obtained from high concentration of cow dung mixed treatments. GC–MS analysis shows no phenolic compounds and predominant level of intermediate metabolites such as4,8,12,16-Tetramethylheptadecan-4-olide (7.61%), 2-Pentadecanone, 6,10,14-trimethyl- (5.29%) andMethyl 16-methyl-heptadecanoate (4.69%) during the vermicomposting process. Spectral results indi-cated that Parthenin toxin and phenols can be eradicated via vermicomposting if mixed with appropriatequantity of cow dung.

� 2013 Elsevier B.V. All rights reserved.

Introduction

Vermicomposting is a bio oxidation and stabilization process oforganic materials that involves joint action of earthworms and bac-

teria, but does not undergo a thermophilic stage [1]. Vermicompostis a rich source of macro nutrients (Phosphorus (P), Nitrogen (N),Sulphur (S), Potassium (K)), micro nutrients (Zinc (Zn), Copper(Cu), Iron (Fe), Manganese (Mn)), vitamins and growth hormoneslike gibberellins, which regulate the growth of plants and mi-crobes. Large number of weed plants that grow at an alarming rateand spread very fast in the cultivated lands, pastures, grasslands

P. Rajiv et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 116 (2013) 642–645 643

and forests are also a good source of organic matter [2]. Vermicom-posting is an effective technique for clearance of animal wastes,crop residues and agro-industrial wastes using low energy [3]. Tra-ditionally, earthworms have been used in domestic compost heapsfor breaking down organic wastes to produce better quality com-post. It is generally known that the epigeic species Eudrilus euge-niae has greater potentiality for degrading organic wastes [4] andserves as the most suitable species for degradation of organicwastes and production of vermicompost in South India [5]. Parthe-nium hysterophorus L. (family Asteraceae) is one of worst weeds inthe world [6]. It is a poisonous, pernicious and aggressive weed.Parthenium is known to badly affect crop production, biodiversity,animal husbandry, human health and even ecosystem integrity [7].It has some water soluble allelopathic chemicals such as phenolicacids and parthenin—a sesquiterpene lactone of pseudoguanolidenature in various parts of the weed [8].

Hsu and Lo [9] suggested that Fourier-transform infrared spec-troscopy is considered a reliable technique for determination ofcompost maturity. Smidt and Meissl [10] investigated throughFT-IR analysis that the reduction in the methylene bands wasdue to decrease in CH2 and CH3 groups suggesting the decompo-sition of aliphatic compounds. GC–MS analysis of vermicompostgives a clear picture of the compounds and chemical compositionlevels.

The aim of present study is to examine the chemical composi-tion of Parthenium mediated vermicompost using FT-IR spectralanalysis technique and Gas chromatography–mass spectroscopyanalysis.

Methods

Raw materials and earthworms

Fresh and healthy P. hysterophorus L. before flowering stage werecollected from in and around fallow lands of Coimbatore, TamilNadu, India. Fresh cow dung was collected from farm houses,Pollachi (T.K), Coimbatore, Tamil Nadu, India. These weeds werechopped into small pieces. The inoculums were essentiallycomposed of weeds and cow dung. Healthy and young E. eugeniaeearthworms were collected from Non-Conventional Energy andRural Development centre, Vadavalli, Coimbatore, Tamil Nadu,India.

Fig. 1. FT-IR spectra of initial feed mixture and vermicompost obtained from 100%Parthenium.

Chemicals and Vermicomposting – experimental setup

All the chemicals (solvents and reagents) were purchased fromsigma–Aldrich chemicals, India. Laboratory glass wares weresoaked overnight in acid cleaning solution and washed thoroughlyin tap and deionised water. Milli-Q water was used for all the anal-ysis. This study was carried out with three replications at Karpa-gam University campus, Coimbatore (11�160N; 76�580E), TamilNadu from March to Ju1y, 2012. The inoculums were preparedby mixing chopped Parthenium and cow dung at different ratios(w/w). The treatments were named as T1 (100% Parthenium), T2

(70% Parthenium + 30% cow dung), T3 (60% Parthenium + 40% cowdung) and T4 (40% Parthenium + 60% cow dung). Mixed compostingredients were added and put into separate tank (1 m depths).Treatments were allowed for pre-digestion for 60 days. A thoroughturning was made after 15 days. After pre-digestion equal size,healthy and unclitellated 100 earthworms (E. eugeniae) were inoc-ulated in each treatment. The tanks were covered with jute bags onthe top to maintain proper heat and humidity. Moisture contentwas maintained by sprinkling water. After 45th day, the vermi-compost samples were collected and stored in plastic containersat cool condition for analysis.

FT-IR analysis of Parthenium mediated vermicompost

The functional groups in vermicompost and initial feed mix-tures were analysed by using Fourier trans-form infrared spectros-copy (Perkin–Elmer 1725x). Vermicompost or initial feed mixturewas dried in hot air oven and five gram of vermicompost was dis-solved in methanol–water (9:1) (v/v), and kept in shaker for over-night at room temperature. After incubation period, the samplewas filtered by using filter paper (Whatman No. 42, Maidstone,England). The extract was dried in hot air oven and pellet was col-lected. The Fourier-transform infrared (FT-IR) spectra were re-corded (wave number ranged 4000–400 cm�1) in mid infraredarea.

GC–MS analysis of Parthenium mediated vermicompost

Five gram of vermicompost or initial feed mixture with 100 mLmethanol–water (9:1) (v/v), in 250 mL was sterilized conical flaskand the conical flask was kept in shaker for overnight at room tem-perature. After incubation period the mixture was filtered by usingfilter paper (Whatman No. 42, Maidstone, England). The extractwas dried in hot air oven and the pellet was collected. Vermicom-post extract sample was analysed on Clarus� 500 Gas Chromato-graph (GC) from PerkinElmer and comprising of Aoc-20i autosampler and gas chromatograph interfaced to a mass spectrometer.The detailed GC–MS experimental conditions have been followedaccording to Gómez-Brandón et al. [11].

Results and discussion

Evaluation of chemical composition by FT-IR spectroscopy

FT-IR spectroscopy was mainly used for analysis of functionalgroups and chemical structure of the material. The FT-IR spectraof initial feed mixtures (compost) and vermicompost are given inFigs. 1–4, respectively. The spectra show several bands, whichare referred to as functional groups of metabolites and other com-ponents, which are used for comparison of degradation and stabil-ization processes. The spectrum of initial feed mixture (T4) showedbroad peak 3396 cm�1, which refers to phenols and alcohol groups(hydrogen-bonded O–H Stretch). Broad peaks of 1730 and1732 cm�1 indicate unsaturated sesquiterpene lactones. Iranshahiet al. [12] reported that absorption of carbonyl function appearedat 1730–1780 cm�1 (saturated and unsaturated sesquiterpene lac-tones). These results are very similar to Iranshahi et al. [12]. A

Fig. 2. FT-IR spectra of initial feed mixture and vermicompost obtained from 70%Parthenium + 30% cow dung.

Fig. 3. FT-IR spectra of initial feed mixture and vermicompost obtained from 60%Parthenium + 40% cow dung.

Fig. 4. FT-IR spectra of initial feed mixture and vermicompost obtained from 60%cow dung + 40% Parthenium.

644 P. Rajiv et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 116 (2013) 642–645

broad peak at 3402 cm�1 indicates carboxylic groups (hydrogen-bonded OAH Stretch). A peak at 2935 cm�1 denotes alkane groups(HACAH Asymmetric & Symmetric Stretch). A peak at 1637–1641 cm�1 a represents amide groups (C@O Stretch and NAHBend). Nitro groups (N@O Bend) is indicated by a broad peak of1381 cm�1. A peak at 1047–1091 cm�1 indicates ether, ester andpolysaccharides groups (CAO Stretch). These spectra peaks werepredominantly obtained in all treatment of vermicompost. The ses-quiterpene lactone functional groups were not seen in T2, T3 and T4

(Figs. 2–4) which show that the vermicomposting process com-pletely degraded the phenols and sesquiterpene lactones. The T1

(100% of Parthenium [without cow dung]) vermicompost spectrum(1778 and 1762 cm�1) shows the sesquiterpene lactone functionalgroups. The T1, T2 and T3 vermicompost spectrum shows peak of3437 cm�1, which indicates the phenol groups. Gupta and Garg[13] have reported that vermicomposting process reduced in bandheight at 3100–3600 cm�1 regions in FT-IR spectra of vermicom-post of cow dung as compared to raw cow dung. Li et al. [14]and Busato et al. [15] have reported that vermicomposting processcaused the disappearance of easily biodegradable compounds and

enhanced the increase of aromatic compounds, which wasconfirmed by FT-IR analysis.

GC–MS analysis of Parthenium mediated vermicompost

There are 50 peaks obtained from GC–MS analysis of initial feedmixture of 40% Parthenium + 60% cow dung and tabulated inSupplementary information A1. A GC–MS spectrum of initial feedmixture (40% Parthenium + 60% cow dung) is show in Supplemen-tary information A2. The maximum peak area was shown by -(+)-Ascorbic acid 2,6-dihexadecanoate (15.54%) followed by 2-Hexade-cen-1-Ol, 3,7,11,15-Tetra (7.69%), 9,12,15-Octadecatrienoic acid,(Z,Z,Z)- (6.13%) and Norethindrone (5.62%). The other peaksindicated a peak area less than 4%.

The results of the GC–MS analysis of vermicompost obtainedfrom 40% Parthenium + 60% cow dung and 49 peaks are tabulatedin Supplementary information A1. A GC–MS spectrum ofvermicompost (40% Parthenium + 60% cow dung) is show inSupplementary information A3. The major peak area is depictedas 4,8,12,16-Tetramethylheptadecan-4-olide (7.61%) followed by2-Pentadecanone, 6,10,14-trimethyl- (5.29%), Methyl 16-methyl-heptadecanoate (4.69%), 6,9,12,15-Docosatetraenoic acid, methyl(4.54), Palmitaldehyde, Diallyl Acet (4.51%), Hexadecanoic Acid,Methyl Es (4.43%), Methyl 5,13-docosadienoate (4.11%), Phthalicacid, isobutyl 2-(2-methoxyethyl) (4.08%) and Bacteriochloro-phyll-c-stearyl (4.07%). The other peaks showed a peak area lessthan 3%. These may be the by-products of the toxins (sesquiter-pene lactones and phenols). The vermicompost and initial feedmixture are rich in ester, aliphatic compounds and alkanes whichmay play vital role in maintaining the nutritional level in vermi-compost. Aqueous extracts of cattle waste derived vermicompostshowed presence of significant amounts of indole-acetic-acid(IAA), gibberellins and cytokinins, which was confirmed by Gaschromatography–mass spectroscopy (GC–MS) [16].

Conclusions

The FT-IR spectroscopy of the vermicompost showed reductionin toxin compounds (sesquiterpene lactones and phenols) duringthe vermicomposting process. GC–MS method is a powerfultechnique for analysis of intermediated compounds and toxincompounds analysis in vermicompost and compost samples. The

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aromatics, aliphatics, alcohols, phenols and polysaccharides aresignificantly decreased while nutritional levels increased throughvermicomposting.

Acknowledgement

We thank to Management of Karpagam University, Coimbatore,Tamil Nadu, India for providing necessary facilities to carry out thiswork.

Appendix A. Supplementary material

Supplementary data associated with this article can be found, inthe online version, at http://dx.doi.org/10.1016/j.saa.2013.08.012.

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