Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 135 (2015) 252–255

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Spectrochimica Acta Part A: Molecular andBiomolecular Spectroscopy

journal homepage: www.elsevier .com/locate /saa

Spectroscopic analysis of vermicompost for determination of nutritionalquality

http://dx.doi.org/10.1016/j.saa.2014.07.0111386-1425/� 2014 Elsevier B.V. All rights reserved.

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

M. Subhash Kumar a, 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

� Eichhornia crassipes is a noxious,aquatic and intransigent weed.� Eichhornia was used as a raw material

for production of vermicompost.� Humic substances functional groups

were analysed by FT-IR spectroscopy.� GC–MS is used to analysis the

compounds of vermicompost andcompost.

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

FT-IR spectra of vermicompost obtained from 50% E. crassipes + 50% cow dung – 10th day.

a r t i c l e i n f o

Article history:Received 21 February 2014Received in revised form 21 June 2014Accepted 2 July 2014Available online 10 July 2014

Keywords:CompostEichhorniaFT-IRGC–MSVermicompost

a b s t r a c t

Spectroscopic analysis has been carried out to examine the compost quality, maturity and nutritional lev-els of vermicompost and compost of Eichhornia. 50% Eichhornia crassipes and 50% cow dung mixtureswere vermicomposted using earthworms (Eudrilus eugeniae) and collected on different days’ time inter-vals. Fourier transform infrared spectroscopy (FT-IR) spectra reveal the presence of humic substance fromcompost and vermicompost, which improves the soil fertility. Gas chromatography–mass spectroscopy(GC–MS) analysis shows maximum level of Benzene propanoic acid (95.98%) and by 2-Propanone, 1-Phe-nyl-, OXIM (10.10%) from vermicompost through earthworms activity. Atomic absorption spectroscopy(AAS) results reported high level of micronutrient from Eichhornia mediated compost and vermicompost.

� 2014 Elsevier B.V. All rights reserved.

Introduction

Vermicomposting is an inexpensive biotechnology tool for recy-cling of a variety of wastes from different nature through the jointaction of earthworms and microorganisms [1]. Various vermicom-posts were produced from different raw materials like cattle

manure, pig manure, agriculture waste and food waste [2].Vermicompost has rich microbial communities significant toincrease soil fertility. As a result of microbial populations, the macroand micro nutrients are in available forms such as nitrates, phos-phates, and exchangeable calcium and soluble potassium [3,4].

FTIR spectroscopy is used to determine composting processesand to describe compost maturity [5–7] or to characterize humicsubstances from compost [8–10]. Hsu and Lo [11] proposed thatFourier-transform infrared spectroscopy is considered a reliable

Fig. 1. FT-IR spectra of initial feed mixture obtained from 50% E. crassipes + 50% cowdung – 0th day.

Fig. 2. FT-IR spectra of vermicompost obtained from 50% E. crassipes + 50% cowdung – 10th day.

Fig. 3. FT-IR spectra of vermicompost obtained from 50% E. crassipes + 50% cowdung – 20th day.

M. Subhash Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 135 (2015) 252–255 253

technique for determination of compost maturity. Castaldi et al.[12] examined the evolution of organic matter during the decayingprocess of municipal solid waste on the basis of changing humicacids. Smidt and Meissl [13] studied through FT-IR analysis thatthe reduction in the methylene bands was due to decrease inCH2 and CH3 groups suggesting the decomposition of aliphaticcompounds. GC–MS analysis of vermicompost gives a clear repre-sentation of chemical composition and compounds.

Water hyacinth (Eichhornia crassipes) is a noxious, invasive,aquatic and intransigent weed [14]. This weed blocks rivers, water-ways and entire lakes and obstructs electricity generation, irriga-tion, navigation, and fishing. It facilitates proliferation of diseases

like bilharzia, kills aquatic life endangering the livelihoods ofmillions of poor people in the tropics, and is now considered a seri-ous threat to biodiversity [15]. This weed is used for wastewaterstreatment [16], and after its wastewater treatment the final dis-posal of E. crassipes is still an unsolved problem.

The aim of present investigation was to improve a consistentand easily manageable method for the determination of compostquality by spectroscopic analysis (FI-IR, GC–MS and AAS).

Materials and Methods

Collection of plants, earthworms and chemicals

Fresh and Healthy E. crassipes plants were collected from Kuri-chi pond, Coimbatore, Tamil Nadu, India (11�160N; 76�580E) in2013. Fresh urine free cow dung was collected from farm houses,Pollachi (T.K), Coimbatore, Tamil Nadu, India. Healthy and youngEudrilus eugeniae earthworms were collected from Non-Conven-tional Energy and Rural Development centre, Vadavalli, Coimba-tore, Tamil Nadu, India. All the chemicals were obtained fromSigma–Aldrich chemicals, India.

Experimental setup of vermicomposting

This study was carried out with three replications at KarpagamUniversity campus, Coimbatore (11�160N; 76�580E), Tamil Nadufrom October–December, 2013. The inoculums were prepared bymixing chopped 50% E. crassipes and 50% cow dung (w/w). Mixedraw material ingredients were added and put into separate tank(1 m depths) and allowed for 30 days for pre-digestion. A thoroughturning was made after 15 days. After pre-digestion healthy, equalsize and unclitellated 100 earthworms (E. eugeniae) were inocu-lated. The 65–75% of moisture content was maintained by sprin-kling water. The vermicompost samples were collected ondifferent days intervals (10, 20, 30 and 40th day) and stored inplastic containers at cool condition for further analysis.

FT-IR analysis of Eichhornia mediated vermicompost

Vermicompost samples were dried in hot air oven and five gramof vermicompost was dissolved in methanol–water (9:1) (v/v), andkept in shaker overnight at room temperature. After incubationperiod, the sample was filtered by filter paper (Whatman No. 42,Maidstone, England). The extract was collected. The Fourier-trans-form infrared (FT-IR) (Perkin–Elmer 1725�) spectra were recorded(wave number ranged 4000–400 cm�1) in mid infrared area.

GC–MS analysis of Eichhornia mediated vermicompost

Five gram of initial feed mixture or vermicompost with 100 mL(9:1) (v/v) methanol–water, in 250 mL was sterilized in conicalflask and was kept in shaker overnight at room temperature. Afterincubation period the mixture was filtered by using filter paper(Whatman No. 42, Maidstone, England). The extract was collected.Vermicompost extract sample was analysed on Clarus� 500 GasChromatograph (GC) from PerkinElmer and comprising of Aoc-20i auto sampler and gas chromatograph interfaced to a massspectrometer. The detailed GC–MS experimental conditions havebeen followed according to Gómez-Brandón et al. [17].

Atomic absorption spectrophotometer (AAS) analysis of Eichhorniamediated vermicompost

Zinc, iron, copper and manganese (micro nutrients) contentswere analysed according to Jackson [18] using diethylene penta

Fig. 4. FT-IR spectra of vermicompost obtained from 50% E. crassipes + 50% cowdung – 30th day.

Fig. 5. FT-IR spectra of vermicompost obtained from 50% E. crassipes + 50% cowdung – 40th day.

254 M. Subhash Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 135 (2015) 252–255

acetic acid (DTPA) and an absorption spectrophotometer (AAS-Shi-madzu AA-700). The experiment was conducted twice and one-way ANOVA was used to analyse the significant differences amongdifferent time intervals for studied parameters. Tukey’s test wasperformed to identify the homogeneous type of the treatmentsfor their dissimilar properties. All the data were analyzed by SPS

Table 1Comparison of GC–MS analysis on compost and vermicompost obtained from 50% Eichhor

Compost of 50% Eichhornia + 50% cow dung Verm

Peak# Retention time Area% Height% Name Rete

1 7.130 0.13 0.30 Acetonyl decyl ether 5.72 8.908 2.80 4.81 Nonane, 3,7-dimethyl- 7.53 9.823 0.20 0.33 Disulfide, diheptyl 8.54 10.401 0.21 0.49 Butane, 1-isocyano- 8.75 13.301 0.85 1.77 1-Tetradecanol 16.06 13.994 1.12 1.82 Tetracyclo[4.1.0.02,4.03 16.97 15.825 4.91 4.30 1-Tetradecanol 17.58 15.985 22.17 13.59 Benzene, 1,2,4-trimetho 17.69 16.212 6.22 5.30 1,2,4-Trimethoxy-5-[(1E) 17.710 17.136 11.03 7.66 1-Dodecanamine, N,N-D 18.211 17.250 3.02 2.38 1,4-Naphthalenedione 20.412 17.339 2.51 2.31 Phenol, O-nonyl- 20.613 18.043 0.92 1.26 Hexanoic acid, 4-meth –14 19.174 4.05 1.99 3-Hexanone, 1-(2,5,6,6-TE –15 19.276 4.02 2.95 (1S,2S,3S)-2-allyl-2-(hyd –16 19.468 18.52 20.50 9-Octadecenoic acid –17 19.724 7.59 12.73 Methyl ester of 3-(3,5-D –18 19.925 �1.01 �0.29 7-Ethylidene-6B,7,8,8A-T –19 21.135 5.26 7.65 7-Tetradecyne –20 21.182 6.04 8.52 9-Octadecenoic acid (Z –21 21.417 �0.56 �0.36 2,3-Diethoxybutane –

16.0 software. Probability levels used for statistical significancewere P < 0.05 for all tests.

Results and discussion

Determination of chemical composition by FT-IR spectroscopy

The analysis of the FTIR spectra of the vermicompost and com-post extracted from the Eichhornia plant at different day intervals(0, 10, 20, 30 and 40th day) and FT-IR spectrum are given Figs. 1–5, respectively. All the spectra bands referred to as functionalgroup of primary and secondary metabolites, which clearly showthe degradation and stabilization processes of compost and vermi-compost. The spectrum showed band at 603 cm�1 correspondingto metal–oxygen (M–O). Broad peaks of 1026 and 1098 cm�1 indi-cate CAOAC stretching of carbohydrates. Broad peaks at 1219,1211 and 1111 cm�1 indicates CAOH stretching of aromatic groupsand CAOAC stretching of aryl ethers and phenols and alcohol func-tion vibrations. A peak at 1373–1411 cm�1 represents anti-sym-metric COOA stretching and aliphatic CAH deformation. Alcoholsand carboxyl functions and NAH vibrations from amides andamines are indicated by a broad peak from 3300 to 3500 cm�1. Apeak at 1047–1091 cm�1 indicates ether, ester and polysaccharidesgroups (CAO Stretch). All the functional groups determine thehumic substances level in compost and vermicompost of Eichhor-nia. The bands at 3420–3400 cm–1 increased while those at 2925,2868–2850, 1460–1440 and 1400–1380 cm–1 were strongly atten-uated during composting indicating a reduction of the aliphaticcontent [19]. Li et al. [20] and Busato et al. [21] have informed thatvermicomposting process produced the disappearance of easilybiodegradable compounds and enhanced the increase of aromaticcompounds, which was confirmed by FT-IR analysis. Gupta andGarg [22] have stated that vermicomposting process reduced inband height at 3100–3600 cm�1 regions in FT-IR spectra of vermi-compost of cow dung. Ravindran et al. [23] described that the com-plete mineralization of polypeptides, polysaccharides, aliphaticmethyl groups and lignin, and formation of a deep nitrate bandin vermicompost compared to compost. Vermicompost has signif-icant level of nitrogen rich compounds and low level of aliphatic/aromatic compounds as compared to the initial level of the biowaste materials, which was confirmed by FT-IR analysis [24]. Rajiv

nia + 50% cow dung.

icompost obtained from 50% Eichhornia + 50% cow dung mediated compost

ntion time Area% Height% Name

12 1.45 1.87 Oxirane, (bromomethy57 �19.90 6.46 Dodecane, 1,1-difluoro75 4.39 2.58 Oxirane, (bromomethy45 1.53 1.98 Oxirane, 2,2’-[oxybis(ME10 10.10 2.62 2-Propanone, 1-Phenyl-, OXIM16 �1.62 0.16 Isoxazolidine04 3.32 1.78 1,3-Butadiene-1,1,4,4-D406 0.06 1.08 O-Toluidine, N-ethyl-22 8.66 4.31 Ethanone, 2-ethoxy-1,46 6.77 4.53 Benzenamine, N-ethyl-2-methy09 �10.73 3.76 Onanoic acid, methyl84 95.98 68.86 Benzenepropanoic acid

– – –– – –– – –– – –– – –– – –– – –– – –– – –

Table 2Micro nutrients level of compost and vermicompost (Mean ± SD, n = 3).

Days Micro nutritional levels (ppm)

Zinc Copper Manganese Iron

0th day 210.00 ± 0.20a 52.13 ± 0.07a 126.10 ± 1.20a 71.03 ± 0.12a

10th day 237.33 ± 1.00b 87.20 ± 0.15b 151.33 ± 1.15b,c 74.16 ± 1.00b

20th day 258.67 ± 0.05c 92.08 ± 0.21c 154.00 ± 1.00b,c 75.36 ± 0.15c

30th day 266.36 ± 0.21d 95.53 ± 1.17d 160.00 ± 0.50b,c 78.45 ± 0.57d

40th day 268.00 ± 2.00e 96.86 ± 0.54d 162.33 ± 1.57d 79.10 ± 0.01d

Mean followed by different lower case alphabets differing significantly (P < 0.05)as established by ANOVA (Turkey’s test).

M. Subhash Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 135 (2015) 252–255 255

et al. [25] stated that toxic compound of parthenin formParthenium mediated vermicompost was characterized by FT-IRanalysis.

GC–MS analysis of Eichhornia mediated compost and vermicompost

There are 21 peaks found from GC–MS analysis of compost of50% Eichhornia + 50% cow dung and is tabulated in Table 1(Supple-mentary material A1). The maximum peak area was shown by ben-zene, 1,2,4-trimetho (22.17%) followed by 9-octadecenoic acid(18.52%) and 1-dodecanamine, N,N-D (11.03%). The other peaksindicated a peak area less than 10%. The results of the GC–MS anal-ysis of vermicompost obtained from 50% Eichhornia + 50% cowdung and 12 peaks are tabulated in Table 1 (Supplementary mate-rial A2). The major peak area is represented as Benzene propanoicacid (95.98%) followed by 2-Propanone, 1-Phenyl-, OXIM (10.10%).The other peaks showed a peak area less than 10%. Rajiv et al. [25]reported that vermicompost has no phenolic compounds andpredominant level of intermediate metabolites such as 4,8,12,16-Tetramethylheptadecan-4-olide (7.61%), 2-Pentadecanone, 6,10,14-trimethyl- (5.29%) and Methyl 16-methyl-heptadecanoate(4.69%) during the vermicomposting process. Aqueous extracts ofcattle waste derived vermicompost showed significant amountsof plant growth hormones, which was studied by Gas chromatog-raphy–mass spectroscopy (GC–MS) [26].

Atomic absorption spectrophotometer (AAS) analysis of Eichhorniamediated vermicompost

The micro nutrients (Zn, Fe, Mn and Cu) level was significantlyincreased in vermicompost as compared to compost. The differ-ence in the micro nutrient level of the vermicompost obtainedfrom different day intervals were significant (P < 0.05) (Table 2).Yadav and Garg [27] stated that 75% of cow dung mixed treat-ment’s vermicompost has great level of micro nutrients Fe(1308 mg kg�1) and Zn (373 mg kg�1). Rajiv et al. [28] reportedthat the maximum concentrations of micro nutrients were in 30%P. hysterophorus + 70% cow dung mediated vermicompost.

Conclusions

Spectroscopic studies (FT-IR, GC–MS and AAS) are an appropri-ate analytical device for compost quality determination and

composting process. FT-IR spectroscopy of the vermicompostshowed humic substances and its functional groups through thevermicomposting process. GC–MS analysis is an important methodfor investigation of intermediate compounds from raw materialsconversions during composting and vermicomposting. AAS analy-sis can be used for examination of micro nutrition level from ver-micompost and compost.

Acknowledgements

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.2014.07.011.

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