Pharmacy & Pharmacology International Journal Research Article Open Access Phytochemical screening, antimicrobial and antioxidant activity determination of Trigonella foenum-graecum seeds Abstract Volume 7 Issue 4 - 2019 To evaluate the phytochemical constituents by maceration and Soxhlet extract, Chua Yoke Hwa,1 Nabila Perveen,1 Neeraj antimicrobial and antioxidant scavenging activity in the seed of Trigonella foenum- Paliwal,2 Naeem Hasan Khan1 graecum. In the present research, maceration and Soxhlet extraction were performed to 1Department of Pharmaceutical Chemistry, Faculty of Pharmacy, the seed of Trigonella foenum-graecum by using 95% ethanol. Phytochemical analysis for AIMST University, Malaysia the important chemical constituents from ethanolic extract was carried out. Antimicrobial 2Department of Pharmaceutical Technology, Faculty of Pharmacy, activity of Trigonella foenum-graecum extract was carried out using agar well diffusion AIMST University, Malaysia method by the zone of inhibition on nutrient agar. MIC and MBC tests were carried out to determine the relatively antimicrobial properties of the extract. Antioxidant activity of Correspondence: Naeem Hasan Khan, Department of Trigonella foenum-graecum was investigated using DPPH free radical scavenging activity Pharmaceutical Chemistry, Faculty of Pharmacy, AIMST assay and total phenolic content test to determine the total amount of phenolic compound University, 08100 Bedong, Kedah D.A., Malaysia, Tel 0060 16 of the seed extract. For phytochemical analysis, test for alkaloids, test for glycosides, test 9372470, Email for saponin, test for Salkowski test, test for anthraquinone, test for tannins, test for phenol, test for carbohydrates that showed positive results for Trigonella foenum-graecum extract Received: July 11, 2019 | Published: August 06, 2019 while the extract showed negative results towards the flavonoid test and reducing sugar test. For agar well diffusion method of antimicrobial activity screening, all the concentrations of Trigonella foenum-graecum showed negative results in all the bacteria except for Bacillus subtilis. The concentrations of extract should increase in order to obtain better results. The antioxidant analysis, total phenolic contents and DPPH radical scavenging showed positive antioxidant results. Keywords:Trigonella foenum-graecum, agar well diffusion method, bacterial strains, inhibition, anticancer Abbreviations: DPPH, 2,2-diphenyl-1-picrylhydrazyl; TPC, such as volatile oils and alkaloid, flavonoid and phenol were reported total phenolic contents; MIC, minimum inhibitory concentration; to have the tendency to exert effect on the bacteria, fungi and parasites MBC, minimum bactericidal concentration; BHT, butylated (Table 1).3 hydroxytoluene Introduction Trigonella foenum-graecum L. also known as Fenugreek which is herb that looks similar to clover. It is a legume and its leaves consist of three small oblong or obovate leaflets. It belongs to the plant family of Fabaceae and grows natively in an area of Iran and Northern India. However, it is now cultivated universally in the area of North and East Africa, China, Ukraine and Greece. Fenugreek seeds are Figure 1 & Figure 2 Trigonella foenum-graecum plant with flower and seeds. commonly used as spices in the cooking for seasoning worldwide, 2 while its leaves can be eaten as green leafy vegetables in the daily diet. Table 1 Taxonomical classification of Fenugreek Fenugreek seeds have been in use for over 2500 years. Its seeds taste Taxanomy Classification bitter, aromatic and are recognized for their therapeutic properties in some diseases such as diabetes mellitus and hypertension. India Kingdom Plantae is the major producer of fenugreek and main consumer for culinary Subkingdom Tracheobionta and medicinal uses.1 The plant and seeds are shown in Figures 1 and 2 respectively. Taxonomical classification of Fenugreek is mentioned Superdivison Spermatophyta in Table 1.2 Seeds of T. foenum-graecum have been used extensively Division Magnoliophyta to prepare extracts and powders for the use for therapeutic purpose. Class Magnoliopsida According to many researchers, the various medicinal properties of T. foenum-graecum, has been found as antimicrobial, antidiabetic and Subclass Rosidae anticancer. The bioactive constituents of seeds of T. foenum-graecum Order Fabales Submit Manuscript | http://medcraveonline.com Pharm Pharmacol Int J. 2019;7(4):175‒186. 175 © 2019 Hwa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and build upon your work non-commercially. Copyright: Phytochemical screening, antimicrobial and antioxidant activity determination of Trigonella foenum- 176 graecum seeds ©2019 Hwa et al. Table Continues... Taxanomy Classification 10-20 seeds. The plant emits a spicy scent that will remain on the hands after touching. Wild and cultivated varieties exist. Flowers are Family Fabaceae 1 to 2, axillary, sessile, raceme, whitish or lemon yellow that bloom Genus Trigonella L. from June to July. Mild Mediterranean climates are most suitable for the growth of the plant. The plant matures in about duration of Species Trigonella foenum-graecum L. four months. The flowering season for the herb fenugreek is usually The name fenugreek comes from foenum-graecum, meaning midsummer. Fenugreek seeds are about 5mm long and are hard. Greek hay, as the plant was traditionally used to scent inferior hay. These are brownish yellow in colour although the colour may vary The name, Trigonella, is derived from the ancient Greek name, which and angular in shape but some are oblong, some are diamond- shaped, denotes the meaning of ‘three-angled’ and refers to the triangular some are almost cubic, and the sides are about 3mm long. A deep shape of the flowers. The first recorded use of fenugreek is described furrow divides them into two parts. They are available in whole and in an ancient Egyptian Papyrus dated to 1500 B.C. The plant is known dried, or as a dull yellow powder, ground from the roasted seeds. in different names based upon the geographical region as shown in According to the report, the life zone of fenugreek is 8 to 27°C, while Table 2. the annual precipitation is 0.4 to 1.5 meters and a soil pH of 5.3 to 8.2. The plant thrives in rich, well-drained soils under plenty of sunlight. 4 Table 2 Common names of Fenugreek It grows slowly and weakly in cold temperatures and moist soils. As a leguminous plant, fenugreek requires almost no nitrogen fertilizer, Languages Names and the plant can use nitrogen to enrich the soils. As native of the Malay Halba Mediterranean area, the seeds have been used for medicinal purposes for thousands of years. It is undeniable that fenugreek is one of the Chinese 胡芦巴 [hú lú bā], Ku Dou oldest cultivated medicinal plants, and now fenugreek is widely grown Tamil Meti, Vendayam today in the Mediterranean countries, Pakistan, Argentina, France, India, North Africa, and the United States as a food, condiment, Urdu Methi medicinal, dye, and forage plant.5 Arabic Hulba, Hilbeh Experimental Italian Fieno Greco Materials and methods Dutch Fenegriek 500g of seeds of Trigonella foenum-graecum (Fenugreek) were Norwegian Bukkehonrkler weighed and allowed to dry under room temperature for duration of one week. The seeds were placed at shaded area without exposing to Portuguese Feno-grego, Alforva the sunlight to avoid any evaporation of active constituent such as Farsi Sambelil phenol. The seeds were crushed into powder by using the crusher and collected in the conical flasks. Swedish Bockshornklee Maceration extraction of seeds powder using ethanol Indonesian Kelabet, Klabat, Kelabat 95% German Bockshornsamen 250g of seeds powder were weighed and added in 500ml of ethanol Russian Sambala, Pazhitnik 95%. The mouth of the conical flask was filmed with para-film and the flask was covered with aluminium foil to avoid any evaporation of Finnish Sarviapila the ethanol and the active components of the extract. The flask was French Fenugrec, Trigonelle then kept in the dark in the cupboard for one week. The mixture was filtered by using muslin cloth. The filtrate was collected in a conical Spanish Alholva, Fenogrec flask and kept covered with parafilm and aluminium foil. Sanskrit Methika Soxhlet extraction of seeds powder using ethanol 95% Japanese Fenu-guriku The weighed seeds powder was transferred to the porous extraction thimble of the Soxhlet apparatus and 350ml of ethanol was transferred Korean Horopa, Penigurik into the 500ml round bottom flask. The temperature was maintained throughout the extraction process. The duration of Soxhlet extraction Description of the plant process was completed in 24 hours. Decolorization of yellow colour of the seeds powder at the extraction chamber could be seen in the end It is a rigidly upright hairy annual legume which can reach a of the extraction process. The Soxhlet extract was collected from the height of 30 to 60cm. The plant grows to a height of about three round bottom flask and was filtrated using muslin cloth. The filtrate feet, has three parts leaves, the long, slender stems possess tripartite, was transferred to a conical flask. toothed, grey-green obovate leaves which normally 20-25mm long. Trigonella foenum-graecum has long stalked leaves up to 5cm long Concentration of extracts using rotary evaporator stipules triangular, lanceolate, leaflets about 2.5 cm long, obovate to oblanceolate. The root is a mass of fingery