Phytochemical Analysis of Four Herbal Seed Extract and Their Use in Poultry Ration

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ORIGINAL ARTICLE Phytochemical analysis of four herbal seed extract and their use in poultry ration

Darshana B. Bhaisare 1, D. Thyagarajan 2, N. Punniamurthy 3, R. Richard Churchil 4and V. Ranganathan 5

1Division of Avian Nutrition and Feed Technology, Central Avian Research Institute, Izatnagar, Bareilly-243122, Uttar Pradesh, India. 2Director of Distance Education, Madhavaram Milk Colony, TANUVAS, Chennai-600 051, India. 3Professor and Head, Veterinary University Training and Research Centre, TANUVAS,Thanjavur-613 403, Tamil Nadu, India. 4Professorand Head, Instructional Livestock Farm Complex, Veterinary College and Research Institute, TANUVAS, Orthanadu, Tamil Nadu, India. 5Associate Professor, Veterinary College and Research Institute, TANUVAS, Orathanadu, Thanjavur-614625, Tamil Nadu, India.

Abstract Hydroalcoholic extract from four herbal seeds namely, thyme (Thymus vulgaris), fennel ( Foeniculum vulgare ), Cumin ( Cuminum cyminum ) and fenugreek ( Trigonella foenum graecum ) were prepared using soxhlet apparatus and concentrated under reduced pressure in a rotary *Corresponding Author: evaporator. The hydroalcoholic extract of above selected seeds were analysed for comparative studies on presence of various phyto-constituents . Darshana B. Bhaisare (PhD scholar) The qualitative phyto-chemical analysis of above four seed extracts

revealed the presence of tannin, phenol, steroids, terpenoids, amino acids, E-mail: [email protected] glycosides and cardiac glycosides. Saponins and flavonoids are present in all seed extracts except cumin, while hydrolysable tannins are present only Received: 30/05/2014 in cumin. Volatile oils are present in thyme and fenugreek seed extract. The

Revised: 25/06/2014 above extracts were subjected for the measurement of total phenolic content (TPC) by Folin-Ciocalteu method using gallic acid standard and the results Accepted: 27/06/2014 revealed the presence of total phenol content of 0.577mg gallic acid equivalent per gm dry weight of thyme extract, 0.528mg per gm dry weight of cumin extract, 1.563mg per gm dry weight fenugreek extract, 1.463mg per gm dry weight of fennel extract. It can be concluded that these phytobiotics can be used as a feed additive in poultry ration.

Keywords: Seed extracts, phyto-chemical analysis, phenol content.

Introduction the prevention and treatment of poultry diseases. Most Antimicrobial resistance is one of the major of the recent research work is being carried out to issues in the world today. It not only affects the poultry search the most safe and economical growth promoter but also harmful for health of the human beings. A as an alternative for antibiotics. In this study, the wide range of feed additives including drugs and qualitative and quantitative phyto-chemical analysis of antibiotics are used in poultry feed to maximize the the four herbal seeds namely cumin, fennel, fenugreek efficiency of production, quality of product and to and thyme, were carried out to assess the usefulness of control diseases. These additives results in residues in their use as poultry feed additives. poultry products, which affect the health hazards to human beings due to various reasons. In the European Materials and Methods Union, the use of antibiotic growth promoters as feed additives is now restricted. Countries such as Australia Phyto-chemical analysis and the USA are following the European example by Analysis of phyto-chemical profile was regulating the use of antibiotics in feeds. In recent performed on four herbal seeds namely cumin, fennel, years, there has been an increased awareness about the fenugreek and thyme, which were procured from the potential compounds of plant origin that have effect in nuts and spices shop located in T-Nagar Chennai. Then

Journal of Poultry Science and Technology | April-June, 2014 | Vol 2 | Issue 2 | Pages 34-37 © 2014 Jakraya Publications (P) Ltd Bhaisare et al...Phytochemical analysis of four herbal seed extract and their use in poultry ration. seeds were later shade dried, powdered and stored in Qualitative phytochemical analysis airtight containers at room temperature for further analysis. This part of the study was conducted at Cumin (Cuminum cyminum ): Qualitative phyto- Veterinary University Training and Research Centre, chemical analysis of hydroalcoholic cumin seed extract Thanjavur. Chemicals, reagents and standards of had bioactive principles like cardiac glycosides, analytical grade were utilized for this study. glycosides, hydrolysable tannins, phenol, steroids, tannin and terpenoids. This study agreed with carrier Qualitative phytochemical analysis study carried out by Madhukar (2013). The hydro-alcoholic extract (Charles et al. , 1993) of the above selected seeds were analysed comparatively for the presence of various phyto- constituents. Chemical tests were carried out using the hydro-alcoholic extract from seeds or powdered specimens using standard procedures to identify the constituents. Qualitative phyto-chemical analysis of herbal seed powder was done following the methodology of Sofowora (1994), which is for amino acids, cardiac glycosides, flavonoids, glycosides, phenol, saponins, steroids, tannin, terpenoids and Fig 1: Calibration curve of gallic acid volatile oils. Fennel (Foeniculum vulgare ): The hydroalcoholic extract of fennel seed subjected to qualitative phyto- Quantitative measurement of Total Phenolic chemical analysis confirmed the presence of phyto- Content with Spectrophotometer constituents such as cardiac glycosides, flavonoids, The above extracts were subjected for the glycosides, phenol, saponins, steroids tannin and measurement of Total Phenolic Content (TPC) by terpenoids. This study was well supported with reports Folin-Ciocalteu method using gallic acid standard of Mandegary et al . (2012) but disagreed with findings which is also known as gallic acid equivalence (GAE) of Tarik et al . (2011) who revealed the absence of method (Singleton et al ., 1999). Gallic acid was used to saponins, essential oil and steroids in ethanolic extracts establish standard curve for total phenolic content. of fennel seeds.

Results Fenugreek (Trigonella foenum graecum ): Phyto- chemical screening of hydroalcoholic extract of Qualitative phyto-chemical analysis fenugreek seed revealed the presence of cardiac Phyto-chemical composition of hydroalcoholic glycosides, flavonoids, glycosides, phenol, saponins, extract of four herbal seeds namely thyme, fenugreek, steroids, tannin, terpenoids, and volatile oils which was fennel and cumin are presented in Table 1. The similar to thyme. This study was in agreement with qualitative phyto-chemical analysis of above four seed Yadav et al. (2011) and Devi and Kumar (2012) while, extracts revealed the presence of cardiac glycosides, differed with Mowla et al. (2009) who reported the flavonoids, glycosides, phenol, saponins, steroids, absence of flavonoids and glycosides in ethanolic tannin and terpenoids, were present in all seed extracts extract of fenugreek seeds. Similarly, Sumayya et al. except cumin while hydrolysable tannins were spotted (2012) who also confirmed the absence of cardiac only in cumin. Volatile oils were traced in thyme and glycosides and volatile oils in aqueous, acid and fenugreek seed extract. alkaline extract of fenugreek seeds.

Quantitative phytochemical analysis Thyme (Thymus vulgaris ): Hydro alcoholic extract of Quantitative estimate of total phenol content in thyme seed extract contained bioactive principles like hydro-alcoholic extract of four herbal seeds is cardiac glycosides, flavonoids, glycosides, phenol, presented in Table 2. The results showed highest total saponins, steroids, tannin, terpenoids and volatile oils. phenol content in thyme and cumin followed by On the contrary, Oyewole et al. (2006) revealed from fenugreek and fennel seeds extract. The gallic acid the qualitative analysis absence of steroids in standard curve with different concentrations is methanolic extract of thyme whereas Dahiya and presented in Fig. 1. Purkayastha (2012) confirmed the absence of flavonoids and steroids in methanolic and ethanolic Discussion extract of thyme.

Journal of Poultry Science and Technology | April-June, 2014 | Vol 2 | Issue 2 | Pages 34-37 © 2014 Jakraya Publications (P) Ltd 35 Bhaisare et al...Phytochemical analysis of four herbal seed extract and their use in poultry ration.

Table 1: Qualitative phytochemical analysis of hydroalcoholic extract of four herbal seeds

Sl. No. Phytochemicals Cumin Fennel Fen ugreek Thyme 1 Cardiac glycosides + + + + 2 Flavonoid - + + + 3 Glycosides + + + + 4 Hydrolysable + - - - tannins 5 Phenol + + + + 6 Saponin - + + + 7 Steroids + + + + 8 Tannin + + + + 9 Terpenoids + + + + 10 Volatile oils - - + + + indicates presence, - indicates absence

Quantitative phyto-chemical analysis weight in methanolic extract, Vadalia et al. (2010) reported 55mg/g in methanolic extract, Chan et al. Cumin (Cuminum cyminum ): The total phenol content (2011) mentioned 4.4mg gallic acid equivalent/100 g in of 1.463mg gallic acid equivalent per gm dry weight aqueous extract whereas Priya et al. (2011) observed was reported in hydro alcoholic cumin seed extract. similar range of 0.876mg of GAE/g of extract in The methanol extract was found to contain different hydroalcoholic (ethanol) extract of fenugreek seed. levels such as 9mg/g dry weight (Thippeswamy and Naidu, 2005), 0.23g/100g of dry weight (Shan et al., Thyme (Thymus vulgaris ): The quantitative estimate 2005), 241.41mg/100g dry weight (Souri et al ., 2008) results reported the presence of total phenol content of while (Chan et al. , 2011) concluded 25mg /100g of dry 1.563mg gallic acid equivalent per gram dry weight of weight in aqueous extract of cumin. thyme extract. In contrary Marghitas et al. (2011) Comparatively low phenol content in all spotted the relatively higher total phenolic content of herbal seeds was observed in the present study which 27mg GAE/g dry weight sample in ethanolic extract of could be due to extract to which phenol content was thyme. estimated. Table 2: Quantitative content of phenol in hydroalcoholic Fennel (Foeniculum vulgare ): The estimated total extract of four herbal seeds phenol content of 0.528mg gallic acid equivalent per gm dry weight in fennel extract was indicated in Hydroalcoholic Total phenol content (mg Gallic extract acid equivalent/g dry wt) results. But variable higher levels of phenol content in Cumin 1.463 different extracts were seen as by Souri et al . (2008) as Fennel 0.528 165.07mg GAE/100g dry weight in methanolic extract Fenugreek 0.577 and by Chan et al. (2011) as 25mg gallic acid equivalent/100 g of dry weight in aqueous extract. Thyme 1.563

Fenugreek (Trigonella foenum graecum ): The total Conclusion phenol content of 0.577mg Gallic acid equivalent per Among the qualitative phyto-chemical tests carried gram dry weight hydro alcoholic fenugreek extract was out, most of the important phyto-chemicals presence was recorded in present study. In contrast with this findings, noticed in all four herbal seed extracts while flavonoids the marked variations were found in total phenol and saponins were absent in cumin while hydrolysable content depending on type of solvent such as Bukhari tannins were spotted only in cumin. However, volatile oils et al. (2008) confirmed 5.65, 6.85, 2.27, 4.04, 1.35 and were traced only in fenugreek seed and thyme extracts. 3.32mg GAE/g dry weight in methanol, ethanol, The higher phenol content was estimated in thyme dichloromethane, acetone, hexane and ethyl acetate followed by cumin seed extract, while lower phenol extract of fenugreek respectively. While, Souri et al . content was found in fenugreek and fennel seed extracts. (2008) indicated higher levels of 194.63mg/100g dry Some of these phytobiotics can be used as a feed additive in poultry ration.

Journal of Poultry Science and Technology | April-June, 2014 | Vol 2 | Issue 2 | Pages 34-37 © 2014 Jakraya Publications (P) Ltd 36 Bhaisare et al...Phytochemical analysis of four herbal seed extract and their use in poultry ration.

Acknowledgement Veterinary University Training and Research Institute, The researchers would like to thank dean of Thanjavur, for providing the necessary facilities Madras Veterinary College, Chennai and Head of required to carry out experiment and their assistance.

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