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International Journal of Chemical Science

International Journal of Chemical Science
Online ISSN: 2523-2843, Print ISSN: 2523-6075
Impact Factor: RJIF 5.22
Volume 1; Issue 2; November 2017; Page No. 110-116

Different extraction methods and antioxidant properties of thyme (Thymus vulgaris L.) herb
Eqbal MA Dauqan, 2 Faten H Thamer, 3 Khalid M Naji, 4 Yahia Alshaibi
School of Chemical Sciences and Food Technology, Faculty of Science and Technology, University Kebangsaan Malaysia,
Bangi Selangor, Malaysia
2, 3, 4
Chemistry Department, Faculty of Science, Sana'a University, Sana'a, Yemen

Medicinal plants have played an essential role in the development of human culture. Medicinal plants are a source of traditional
medicine. Among different species of Thymus, (Thymus vulgaris L.) is used more than other species in therapeutic dosage forms.
In Traditional medicine, Thymus vulgaris L. is cultivated in many countries by most people especially in rural areas depend on
herbal medicines to treat many diseases including inflammation-related ailments such as rheumatism, muscle swelling, insect bites
and pains. Also the modern medicine in essential oil of thyme has demonstrated the compounds have shown antioxidant,
antibacterial and antifungal properties. Thus the highlight of the paper was to review the different extraction methods and
antioxidant properties of Thyme (Thymus Vulgaris) herb.

Keywords: medicinal plants, traditional medicine, Thymus vulgaris L., antioxidants

1. Introduction They are among our oldest medicines and their increasing use
Human body constantly creates free radicals culminating in an in recent years is evidence of public interest in alternatives to
“oxidative stress” when their elimination by antioxidant conventional medicine [11]. Since prehistoric times, herbs have
defense mechanisms is not sufficient [1]. Oxidative stress also been the basis for nearly all medicinal therapy until
contributes to the pathogenesis of many human diseases; synthetic drugs were developed in the 19th century [16].
therefore the intake of antioxidative agents is important for the Herbs are still found in 40% of prescription drugs [16]. In
prevention of chronic diseases [2]. Antioxidants play an addition, herbs are used for many other purposes including
important role in preserving of food too. In food industry beverages such as tea, dyeing, repellents, fragrances Many
widely used synthetic antioxidants as butylated hydroxitoluen recent studies found out that herbs contain various
(BHT) and butylated hydroxianisole (BHA) are very effective phytochemicals including antioxidants [17, 18, 19, 20, 21]. At the
because of their low cost, high thermal stability and efficiency same time it has been shown that Reactive Oxygen Species
but they are instable and they can play role as promoters of (ROS), such as hydroxyl radical, hydrogen peroxide, and
carcinogenesis [3, 4, 5, 6]. Due to these reasons, there is a superoxide anions, are produced as by-products in aerobic
growing interest in the study of natural additives as potential organisms and have been implicated in the pathology of a vast
antioxidants [7]. The presence of antioxidants in many spices variety of human diseases including cancer, atherosclerosis,
gives them food-preserving properties too, especially in diabetic mellitus, hypertension, Aids, aging, cardiovascular
preventing oxidation of lipids [6]. diseases, cataracts, immune system decline and brain
Nevertheless, the use of synthetic antioxidants in the food dysfunction [9]. It was also found out that free radical
industry has been questioned regarding its innocuousness. formation is controlled naturally by various beneficial
Studies about spices and aromatic herbs have been widely compounds, namely antioxidants. There has been lots of
emphasized, can act as an alternative to prevent the oxidative evidence that consuming foods of plant origin (fruits,
deterioration of food and reduce the use of synthetic vegetables, tea, coffee and others) is associated with reduced
antioxidants [8]. The use of natural antioxidants from food incidence of these diseases [19].
plants has the following advantages: They are accepted by the Moreover, knowledge and application of such potential
consumers; they are considered safe; they do not need safety antioxidant activities in reducing oxidative stresses in vivo has
tests; they have functional and acceptable sensory properties prompted many investigators to search for potent and cost-
. Studies found in literature have demonstrated that the effective antioxidants from various plant sources [19]. These
spices belonging to the Lamiaceae family, as well as their research activities have contributed to new or renewed public
extracts and essential oils, are efficient antioxidants [8, 10, 11, 12, interests worldwide in herbal medicines, health foods, and
13, 14, 15]
. nutritional supplements. It is therefore of big interest to
Herbs are usually considered as plants with aromatic systematically check for the presence of antioxidants and their
properties and are mainly used in spicy foods and for antioxidative capacity in medicinal herbs that are widely used
preparation of herbal teas in folk medicine [13]. Medicinal in local folk medicine [ 22].
plants have always been considered as a source of health [16]. Studying medicinal plants helps to understand plant toxicity


signifying courage the plant especially for tea. They are often cultivated because of their aromatic qualities and also of their easy cultivation. responsible for its antioxidative activity [36]. Thyme prefers lightweight. Figure 1 shows the temperature. drying distillation [25. Thyme leaves are terribly little. Locally Thymus vulgaris L species native ornamental purposes [26]. 28]. and is utilized commercially for flowering and used worldwide [31].and long and vary significantly in form and hair covering. Hajah. luteolin. this method gives a greater shrub.1 Steam Distillation 'Argenteus' (silver thyme). herbs. Thyme is the general name for the many shahel. and also antifungal effects [32]. 3. The antiseptic effects are mainly due to horizontal and upright habit. The stems become woody with the presence of volatile oil. condiments.5 to 5 mm carvacrol. this aromatic plant has fumigate either because they used it as incense for its balsamic been used for the preparation of different aliments to cure odor or because it was taken as a type of all sweet smelling various curable and chronic diseases [32]. vulgaris leaves are oval to phenolic acids (rosmarinic acid) [34] and by flavonoids rectangular in form and somewhat fleshy aerial components (quercitin. age [28]. sweeter than sage. etc. Methods of Extraction of Thyme Oils drained soils with a pH of 5. eriocitrin. 2. with a semi ever green As a tea it is useful for colds. Thyme species do best in Michalak [37] reported that the thyme essential oil quality and coarse. 111 . In food Thymus vulgaris L. T. Numerous cultivars and hybrids have been Essential oil of thyme herb has usually been obtained by some developed for ornamental purposes.0.The cultivar 'Silver Queen' possess Steam distillation procedure is widely used for essential oil white-margined leaves [30]. Many species of the family are reported with high phenolic contents and antioxidant capacities [24]. Many of them are commonly used as culinary herbs. depending on the variety. thyme has being held in ancient and medieval days to be a great source antiseptic effect due to the presence after penoidic compounds of invigoration its pleasant qualities inspiring courage [27].1 Thyme (Thymus Vulgaris L) Thyme is one herb of Lamiaceae family. From the medical point of view. It is slightly spicier than oregano and extraction method is very important. Yemen herb varieties of the Thymus species. with every species having a rather The antioxidant activity of wild thyme is given especially by completely different scent. apigenin. and wherever the plants don’t seem thymol are the main phenolic components which are primarily to be shaded. seasonal variations. can be used in fresh or dried form meaning perfume or Thumus. sesquiterpenes (β-cariophilen. Amran. Lamiaceae Family Lamiaceae family is a group of about 210 genera and some (A) (B) 3500 species [23]. conditions. There are two methods use steam distillation: varieties of thyme in different cities in Yemen. Yemen. Others derive the name from the Greek words Thyo. which mainly contains thymol. dry. Thyme doesn’t like excessive wet as a result of its condition it will gets rot diseases [29]. rough soils that may be unsuitable for several yield depend on many factors and choosing a suitable alternative plants [28. It desires full sun to grow to its best potential. Thyme India and Europe thyme grows well during a temperate to contains high concentrations of phenols [34]. simple steam distillation and Clevenger system [38]. It dates back to 3500 BC by Sumerians and Egyptians [25]. well. to the Sultanate of Oman and Yemen known as “zaater” and The name of Thyme in its Greek form was first given to the their dried whole parts are used in herbal tea. all of which are native to Europe and Asia. Sana'a. may affect the composition of thyme. principally by steam Many factors such as harvest time. with a green leaves that seldom grows quite 5 cm tall to or lesser compounds instability under the influence of high extend horizontal about 50cm or more. is one of the most popular hybrid plant principal type. and plant by the Greeks as a derivative of a word which meant to folk medicine. Common or garden thyme is considered the Thymus vulgaris L. germacren D or nerolidol) [33]. Yemen and (C) thyme in Maswar. French. The many cultivars include 3. Its spread to Europe was (C) due to Romans.0 to 8. In Yemen thyme is a seasonal separation. (B) thyme in Al- liqueurs & cheese. Thyme is a tiny perennial shrub. α-pinen [27] and other mono. p-cimen. Nomenclature can be very methods such as: confusing. 30] . Therefore the objective of this paper was to review the different extraction methods with different conditions used to extract thymus oil and antioxidant properties of Thyme (Thymus Vulgaris) herb. sunny climate. Carvacrol and heat. linalool. as they used it to give aromatic flavors to Fig 1: (A)Thyme in Bani-Matar. are used for volatile oil production. usually 2. beyond its efficiency. serpyllin) [35]. German and English varieties vary by leaf shape and color and essential oils. asthma and bronchitis [25] or for groundcover that seldom grows quite 40 cm tall it’s each the treatment of acne. 2. Since ancient times.International Journal of Chemical Science and protect human and animals from natural poisons.

Turkey . . it reaction only occurring in strong hydrogen bond-accepting can also minimize possible degradation because it can operate solvents such as methanol and ethanol. Table 1 shows the assay provides fast and reliable results for plasma. During early work Can offer a good yield and preserve the properties of on DPPH.2%. 4. able to accept an electron or a hydrogen atom creating from a solid matrix using a liquid matrix. 2-diphenyl-1-picrylhydrazyl (DPPH) [20]. Carvacrol. The leafy parts of thyme and its oil are known concentration is directly related to the total ferric utilized in foods for the flavor. it will cause damage complex. The radical extraction is concentrate antioxidant components of raw absorbs visible light at 515 nm appearing red-violet in color. the reaction starts with an electron extraction process and reduces extraction time due to low transfer. researchers to extract phenolics from thyme leaves. A good choice for analyzing both lipophilic and hydrophilic 3. 22. researchers concluded that the reaction takes place antioxidants. thymol and Hydrodist. 48]. while hydrogen atom abstraction is a slow secondary viscosity and relatively high density of supercritical fluid. γ-Terpinene (13.C Ankara. 44. leads to an intense blue-colored ferrous tripyridyl-triazine Once the chain reaction happens in a cell. which obviously invalidate the determination [41]. p-Cymene (18. thymol and Solvent =1. single different methods used for thyme extraction.4±0.8-Cineole 14. Anti-oxidants stop these chain reactions spectrophotometrically [43.33%). 45]. positive results. A drawback is that the extracting fluid in the liquid state during the whole extraction solvent effect has to be quantified carefully. By applying these conditions. Pressurized liquid extraction is radical is a stable molecule and does not need to be generated. In the case of a process. The aim of the diamagnetic molecule DPPH (non radical). The reduction of ferric ions to ferrous ions radicals.2%) and carvacrol illation 0.3 Supercritical Fluid Extraction the appearance of a pale yellow color [41]. the competition of hydrogen abstraction processes result in which typically higher extraction yields are between the antioxidant and the solvent can lead to false obtained with lower volumes of organic solvents [14].8 Cineole (40.C .5% Apinene and 6.1.4 IC50 .International Journal of Chemical Science 3. the FRAP assay is simple and Table 1 used DPPH and FRAP methods to determine the inexpensive.21 ± south of France .4 Pressurized Liquid Extraction (PLE) antioxidants is a 50% (v/v) water/ethanol mixture [42].2 Ferric Reducing Antioxidant Power Assay (FRAP) different molecules. but when it is mixed with a protic solution the reduced form (non radical) is generated with the loss of the violet color and 3. carvacrol extraction (mmol/L HPLC 7. Supercritical carbon dioxide is the transfer-based assays. This is a The DPPH is a very stable and commercially available free process designed to separate soluble compounds by diffusion radical. based on the use of solvents at temperatures above their The results are highly reproducible and comparable to other normal boiling points and pressures enough to keep the antioxidant detection methods [41]. .2% linalool 4. antioxidant antioxidants in pure solution. cannot be used to determine antioxidants containing oxidizable groups [46.6% 1.5 GAE G. 40]. Oxidation is a chemical process that The FRAP assay measures antioxidant capacity by studying transfers electrons or hydrogen from a substance to an the reduction of the complex ferric tripyridyl-triazine (FeIII- oxidizing agent. the formation of which can be followed or death to the cell. As in other electron in absence of light and air. The only drawback of this method is that it antioxidants activity.6% Camphor methanol) Thymol (41. the scavenging activity is strongly most widely used extraction solvent [39]. influenced by the pH and the solvent properties. aroma and preservation and reducing power of the antioxidant in the sample.2%). and mixtures of antioxidants in values and essential oil analysis. [30] q-Cymene. Antioxidant Properties An antioxidant is a molecule that inhibits the oxidation of 4. All the previous studies in aqueous solutions. A relatively recent solvent extraction technique could in The DPPH assay is a valid and easy method for evaluating the principle eliminate some of the drawbacks of the classical scavenging activity of antioxidant compounds because the solvent extraction methods.1 2. The FRAP additionally in folk medicines [25. material [37].12%). In fact.08 µg/mL α-Pinene (13. antioxidant values and essential oil analysis Essential Main Methods of Value of Total Country Ratio of compounds Value of DPPH Oil References compounds extraction FRAP phenols Analysis Thymol 61% %92. Oxidation reactions will produce free TPTZ) at low pH.MS [5] . G.08%). 20. 2-Diphenyl-1-Picrylhydrazyl Radical Assay (DPPH) The DPPH assay measures the antioxidant properties of Table 1: The different methods used for thyme extraction. This method can be used for the extraction of through the transfer of a hydrogen atom between the radical polyphenol from plant tissue and greatly facilitates the and the solution.8% borneol. The difference in absorbance by removing free radical intermediates and inhibit different with respect to a reaction mixture containing ferrous ions of oxidation reactions. faster extraction protic solvent. 4. Moreover. IC50= 4. In turn. these radicals will begin chain reactions.2 Solvent Extraction compounds in reference to their ability to scavenge the radical Solvent extraction was the main method adopted by most anion 2. 47. 112 .

.05% Camphor 9.9–6.46% Carvacrol extraction Carvacrol 3.44 ± AE) . .00 ± affeicaci 9. . . (mg/mL TAE)] AE) nt [11] andQuercetin extractor 0.8 Cineole 7.4±0.0% Bucharest.95% Thymol (30.01 0. .01 69.60mg [IC50(mg/mL trolox/gT Catalonia Soxhlet equivale Caffeic acid .2–4.53%) and thymol distillation HPLC Myrcene 1. Turkey q-Cymene (mmol/L . GAE/100 GAE.12%) .Rosmarinic acid Soxhlet Morocco . and Camphor (0–7.79% β-Myrecene7.92 (mmol 5. p-Cymene (18.40 TAE) Romania Cravacrol Solvent Carvacrol (21–37%) Thymol(10–17%) . =1. . . 1. 70% .1–22.5%). . GC–MS [44] 2.Rosmarinic acid andQuercetin .33%).5 GAE Thymol and Solvent G.44 ± 0. 2000 .2%).8%). [57] and Thymol Distillation GCMS 113 . .05% Camphor carvacrol extraction 9.46% Carvacrol 3.13 g 4. G.20%g-Terpinene 2. Borneol (0. GC.002 . GC–MS [39] Sabinene 1.4–20.60mg [IC50 trolox/gT equivale Caffeic acid.48% Soxhlet Sultanate of method.08%).16 g Valtice Solvent [9] .4 IC50 (0.2–14.48% p-Cymene 1.24% linalool 2.6–7.26% .C (Mugla.48% Supercriticl p-Cymene 1.05% Put in boiling HPLC- Croatia Rosmarinic acid 17.39% Linalool 2.2%).06% γ-Terpinene26.56% Cymene6.International Journal of Chemical Science 1-Octen-3-ol 0. IC50= . .50% Thymol 12–61% Carvacrol 0.45±0. TAE)] 0. [52] water PDA glucoside 2.37±0. GC.6%).73% Borneol 3.05 ±0.03%a-Pinene 1. .MS- 89%. Thymol and Hydro.44 % 482. [53] Algeria extraction mg/g TAE Thymol 47% p-cymene 34% Thymol and Steam Paris (France) Linalool1 .2%).92 (mmol 5. 1-Octen-3-ol 0. . GC-MS [54] p-Cymene distillation Carvacrol4 . .21 Apigenin-7-O.2%) and α-Pinene (13.00 ± acid/g 9.23% Sabinene 1.02 65. Spain fluid . . .21 ± Iran γ-Terpinene (13.40 d/g TAE) Carvacrol (21-37%) Cravacrol Solvent Armenia . Italy . .MS [24] (Fares) and Thymol extraction Cravacrol Steam HS- Ravenna. 76-98% [51] Oman Solvent mg/L extraction phydroxybenzoic acid 0.8-Cineole %92. .a- Pinene (0.86%) Cravacrol steam GC.10 - extraction g−1dw 0 g−1dw 9.C /MS [56] Carvacrol distillation 0.55% Thymol 57. .8 Cineole (40. . . IC50= 4. [8] (Spain) extractor nt caffeic 0. . [50] Romania p-Cymene (30. .16% Caryophyllene 2.30g/l Seva Flora 1. GC-MS [49] and thymol Distillation Phellandrene2.Cravacrol Steam Palestine .1. [55] Carvacrol extraction HPLC (9.63% Sabinene 4.MS [24] Thymol (10–17%) and thymol extraction .23% thymol and Murcia. . .39% Linalool Thymol and Greece fluid .77% 1.08 µg/mL .8 Supercritical Cineole 7.02* 65.55% Thymol 57.3% Thymol (41. Solvent 0. .6%. . Cevacrol0. .91% α-Pinene2. methanol) Linalool (2.07 ± Constantine.44 % 482.77% 1.

extraction.MS- Cevacrol0. Linalool (4.8-Cineole (4. .4) Linayl acetate distillation 3. Natural Antioxidants: Sources. 10(4):221-47.17% .53%) Myrcene 1.10 .50 southern Solvent 92.37±0.73% Borneol 3. HPLC [58] extraction GAE/10 Poland 0 g dw Tthymol (50.8 (0. 2(2):1276-85.3). . GAE. . Comprehensive Reviews in Food Science and Food species around the world due to climate and method of Safety. Miladi H. extraction. 2010. Barry M.Iran . 70% . References Effects of Marjoram and Thyme in Coated Refrigerated 1. 2011. .14%) 1. .3). Mohamed A.2 (0. Yasin NMN. Mohamed A.95% thymol (30.24% Linalool 2.3 Dalmatia. Conclusion 2.05%IC50=0. 5(6):729-39.1 (0.) Extracts.2 (0. Thymol and 1.54 mg Wielkie. Compounds. p-Cymen) 15.06%γ-Terpinene26. .48%) Belgrade. . Antioxidant Effect Research.: Gas 6. GC-MS [59] Serbia p-cymene.8) Terpinene 2. Khaled A S. 2007.4-Terpineol 85.69%). Business. Solvent 76-98% [51] mg/L extraction Phydroxybenzoic acid 0.) and Oregano (Origanum 114 . Thymol 16. Hamdy R.86%) 89%. Brewer MS.5 (0.56% m-Cymene6.6) . . 2010. Donia M.79%).16 g Estahban Solvent . Industrial Crops & Products. A Review on In-Vitro Semi Fried Mullet Fish Fillets. Timothy OB. Patricia VDR. Science. GC-MS [60] (Croatia) linalool 6. [53] extraction mg/g TAE 517 ± Grabowo Solvent 0.World Journal of Dairy & Antioxidant Methods: Comparisions.100 GAE.) is an important medicinal plant Oxidative Stress II. thyme vary widely among the investigated Thymus vulgaris L. 2013. 594(25):395-408. Jorge N. 6. Rosmarinus Officinalis L. Linalool 40. Thymol and Solvent p-Cymene (24.35%) Pinene 2. . 5.91%α-Pinene2.” Natural for research fellow.2 (3. Considerations. .05 ±0. This work was supported by Sana’a University and Institute of Cytotoxicity and Antioxidant Properties and Antibacterial International Education Scholar Rescue Fund (IIE-SRF).35% Fe2+/mm . The antioxidant 3.).International Journal of Chemical Science Phellandrene2. Hussein M. Madhu SC.. extraction γ Terpinene (4.48% Soxhlet and 2000 Saveh. . Also this review showed I.0 (0.3 (0.07 ± Solvent 0. [52] farm-France water 0g/l PDA Apigenin-7-O-glucoside 2. [10] California USA and thymol distillation HPLC p-Cymene (30. Correlations and Food Sciences.2).002 Croatia . ol L-1 á-Terpineol4.2). Iran extraction g−1dw 0 g−1dw 9. but the most components are thymol and cravacrol 4. 2(1):01-09. (µgGAE/ [11] Morocco . Carolina MV. which belongs to the Lamiaceae family. Finally thyme has a very high antioxidant activity 2013. plant. of Thyme (Thymus Vulgaris L. amount of total phenolic and main components of Mechanisms of Action.3 HPLC- Rosmarinic acid 17. extraction - . hydro.45±0.01 1. International Journal of PharmTech 7. Badarinath MR. 43:827-31. Antioxidant and Antimicrobial 7.2 (0. Essential Oil of Thymus Vulgaris L. USA Activities against Foodborne Pathogens. Carvacrol 6. There are a lot of methods for extraction of thyme oil Sage (Salvia Officinalis L. activity.13 g 4. and Marjoram (Origanum but the most used methods are steam distillation and solvent Majorana L.8 (1. mg DM 5. Advanced Protocols in Thyme (Thymus Vulgaris L. Total Phenols and that total antioxidant activity increase with increasing total Phenolic Compounds in Thyme (Thymus Vulgaris L. Acknowledgement Chromatography-Mass Spectrometry Analysis.01 Shambolia Put in boiling 69. Rihab BS.7) 34. .6).21 .79%β- Myrecene7.3). and Potential Applications.0% northern Cravacrol steam GC. Khalel in most Thymus vulgaris L.). .16% Caryophyllene 2. Eglon M. Borneol 1. phenols. . compared with a lot of medical plants. [9] Branch. Evaluation of Antioxidant Activity .63% Sabinene 4. . 1. .

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