CHEMICAL COMPOSITION OF FAGOPYRUM ESCEULENTUM MOENCH SEED THROUGH GC-MSHTML Full Text
CHEMICAL COMPOSITION OF FAGOPYRUM ESCEULENTUM MOENCH SEED THROUGH GC-MS
Neeraj *, Neeru Vasudeva and Sunil Sharma
Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science and Technology, Hisar - 125001, Haryana, India.
ABSTRACT: Medicinal herbs are a precious heritage for us and since long is a part of traditional as well modern system of medicine. These herbs provide less toxic and more effective drug molecules which are helpful in various ailments. In the present work, we have focused on an edible herb which bears a large number of medicinal properties. Fagopyrum esceulentum Moench (Buckwheat) has a long tradition of being used as food and medicine which is endorsed with abundant of nutrients like flavonoids, polyphenols, proteins, vitamins, phytosterols, carotenoids, and minerals, etc. in it. In the present study, chemical and pharmacological investigation of buckwheat seed ethanol extract were carried out for determining its usefulness in human health. The GC-MS analysis was performed and based on Mass Spectra fragmentation pattern and retention indices, phytoconstituents were identified. Major constituents identified were 9-octadecenamide, n-hexadecanoic acid, ethyl linolate, 9-octadecenoic acid (z), 2, 3-dihydroxypropyl ester, ergost-5-en-3-ol (3.beta.24r), gamma-sitosterol, lupeol, fumaric acid. These phytoconstituents show various pharmacological activities like anti-inflammatory, antioxidant, antibacterial, anticancer, anti-hyperglycemic, dyslipidemic and cardiovascular activities.
GC-MS, Fagopyrum esceulentum Moench, Phyto-constituent, Retention indices
INTRODUCTION: Fagopyrum esculentum Moench., commonly known as ‘Buckwheat’ or ‘Kuttu’ is a herb belonging to Polygonaceae family. It can easily grow in poor soil and can bear acidic environment. It mainly grows in the countries in the northern hemisphere of world 1. The composition of buckwheat (carbohydrate, crude fat, crude fiber and crude ash content, etc.) resembles with wheat; therefore it is referred to as pseudocereal 2. Usefulness of buckwheat is attributed to its prosperity in medicinally active components.
Along with its curative potential, it also has many prophylactic effects 3. It is a gluten-free grain which makes it a suitable substitute for celiac disease patients 4. Buckwheat proteins have a unique amino acid composition with several biological activities.
Besides high-quality protein, buckwheat also has several other medicinally active components like starch, trace elements, dietary fibers, phenols, phytosterols, D-Chiro-inositol derivative, imino-sugars, fatty acid, and minerals, etc. 3 Flavonoids which are also known as nature tender drugs are present in a huge amount in buckwheat 5. Consumption of buckwheat reduces risk of diabetes because of its high content of magnesium. It also cures the condition because of chiro-inositol present in it 6, 7. Buckwheat alters cholesterol metabolism thus helpful in avoiding gallstones 8.
The high fiber content and plant lignans make buckwheat helpful in reducing risk related to colon cancer, breast cancer, and cardiovascular problems. Postmenopausal consumption of its grains is beneficial for a person with high cholesterol, high blood pressure, and cardiovascular problems 7. To amplify the use of buckwheat for the improvement of human health, there is a need to further explore its chemical components. GC-MS is a fusion of two analytical techniques namely Gas Chromatography (GC) and Mass Spectroscopy (MS). GC separates different components of the sample into pulses of pure chemicals by flowing an inert gas (mobile phase), which carries the sample through a stationary phase fixed in column 9.
As the sample exits the end of the GC column, it is fragmented by ionization, and the fragments are sorted by mass to form a fragmentation pattern. These spectra of compounds are collected by the mass spectrometer, which identifies and quantifies the components according to their retention behavior and mass-to-charge ratio (m/z) 10. Mass Spectra and retention time (RT) of a component known as the molecular fingerprint are used as the identifying characteristic of that component. Retention time (RT) and the fragmentation pattern (mass spectra) of every component is matched to commercial libraries like NIST and Wiley collections. In the case of complex mixtures, Kovatz retention index (RI) has to be used because Mass Spectral matching alone has been found insufficient for compound matching and recognition. RI is the most popular dependent variable in GC-MS studies because of its excellent reproducibility and accuracy. RI criterion is independent of the chromatographic column conditions and problems caused, during the injection of volatile and thermally unstable compounds, problems in a stationary phase like overloading, bleeding and trapping of the solutes. Present work is a GC-MS analysis of the chemical composition of buckwheat and their corresponding health benefits.
MATERIALS AND METHODS:
Material: F. esculentum seeds procured from local market Hisar, Haryana. Analytical grade ethanol and petroleum ether (60-80 ºC) were purchased from Hi-media, Mumbai, India. Alkane-series was formed Sigma-Aldrich India.
Authentication of Plant: Authentication of the seed was done by Dr. Sunita Garg, Emeritus Scientist, CSIR-NISCAIR, New Delhi, India (letter no. NISCAIR/RHMD/Consult/2016/2984-11). A voucher specimen has been deposited in Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science and Technology, Hisar, Haryana for future reference.
Preparation of Sample for GC-MS: The seeds of F. esculentum were dried under shade and powdered coarsely using pestle and mortar. Coarsely powdered seeds were defatted with petroleum ether (60-80 ºC) and extracted by continuous hot percolation using Soxhlet assembly at room temperature for 7 h with 95% v/v ethanol. The extract was filtered; the solvent was recovered using rotary evaporator at 45 °C under reduced pressure. A semi-solid mass was obtained which was kept in a desiccator for further use.
Instrumental Conditions: Chemical analysis of ethanol extract of buckwheat was carried out using a Shimadzu GC-2010 attached with an autosampler (AOC-20i). Helium (>99.999%) was used as the carrier gas with an Rtx 5 MS capillary column (Restek Company, Bellefonte, USA: cross bond 5% diphenyl / 95% dimethyl polysiloxane) having dimensions 30 m (length) × 0.25 mm (diameter) × 0.25 μm (film thickness). The temperature was programmed from 100 °C (3 min), then further increased to 280 °C at a ramp rate of 10 °C/min (19 min hold). The flow rate of carrier gas was 1.21 ml/min, the linear flow velocity of 40.9 cm/s and the split ratio was 10:1. The data was processed on GC solutions software for composition. GC-MS instrument was equipped with the mass selective detector, having ion source temperature of 230 °C, interface temperature of 260 °C, a solvent cut time of 2.50 min threshold of 1,000 eV and mass range of 40 to 650 m/z. The identity of the components was assigned by comparing their GC retention times with those of authentic samples as well as with known components of the standard composition of the fragmentation pattern with that reported in NIST and Wiley computer libraries.
RESULTS: GC-MS analysis of F. esceulentum seed extract revealed the presence of 42 components Table 1.
TABLE 1: CHEMICAL COMPONENTS ANALYSED THROUGH GC-MS
|Retention Time||Retention Indices||Area
|Name||Molecular Formula||Molecular Weight||Pharmacological
|5.493||959||0.07||2-Heptenal, (E)||C8H16||112||Flavouring agent||13|
|7.013||1039||0.07||2-Pyrrolidinone, 1-methyl||C5H9NO||99||Antioxidant and anticancer||16|
|9.095||1146||0.14||4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl||C6H8O4||144||Anti-inflammatory, analgesic, antibacterial, antifungal||18|
|10.114||1200||0.11||Dodecane||-||-||No activity reported||19|
|13.616||1399||0.17||Tetradecane||C14H30||198||Elephantiasis, asthma, throat disease, bronchitis||21|
|17.861||1677||0.10||Pentadecafluorooctanoic acid, dodecyl ester||C20H25F15O2||582||Antibacterial||25|
|18.756||1742||1.17||2-Ethoxycarbonyl syringic acid||C12H14O7||270||Antidiabetic and
|19.035||1762||0.61||Tetradecanoic acid||C14H28O2||228||Antioxidant, cancer preventive||27|
|19.537||1799||0.29||Octadecane||C18H38||254||Anticorrosion agent, Antiseptic||21|
|23.264||2099||2.56||9-Octadecenoic acid, methyl ester, (E)-||C19H36O2||296||Cancer preventive, Antioxidant||27|
|23.556||2125||0.27||Methyl sterate||C19H38O2||298||Anti-diarrhoeal, anti-proliferative, cytotoxic||27|
|23.973||2162||26.32||Ethyl linolate||C20H36O2||308||Antibacterial agent||30|
|25.330||2285||0.42||Fumaric acid||C17H31NO4||313||Food industry||11|
|25.651||2314||0.12||Palmitic acid monoethanolamide||C18H37NO2||299||Antioxidant, hypocholesterolemic nematicide, pesticide, anti-androgenic flavor, hemolytic, 5-Alpha reductase inhibitor||18|
|26.506||2391||0.28||Eicosanoic acid, ethyl ester||C22H44O2||340||No activity reported||31|
|27.317||2455||1.23||Fumaric acid||C17H31NO4||313||Food industry||11|
|28.093||2509||0.53||Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester||C19H38O4||330
|Pesticide, flavouring agent,
|28.293||2518||0.47||1,2-Benzenedicarboxylic acid||C24H38O4||390||Antimicrobial and antifouling agent||23|
|28.997||2518||0.24||Docosanoic acid, ethyl ester||C24H48O2||368||Detergents, floor
|30.053||2603||13.35||9-Octadecenoic acid (Z)-, 2,3-dihydroxypropyl ester||C21H40O4||356||Anticancer||27|
|30.769||2692||0.43||Heptadecanoic acid, ethyl ester||C19H38O2||298||Antioxidant||27|
|30.912||2711||0.14||Squalene||C30H50||410||Antioxidant, skin hydrant, emollient||33|
|34.621||3240||1.24||Ergost-5-en-3-ol, (3.BETA.,24R)-||C28H48O||400||Dyslipidemia and
|34.910||3265||0.29||Stigmasterol||C29H48O||412||Antioxidant, hypoglycemic and progesterone precursor, antimicrobial, anticancer, anti-arthritic, anti- asthamatic, anti-inflammatory, diuretic||32|
FIG. 1: GC-MS CHROMATOGRAM OF F. ESCULENTUM SEED EXTRACT
Fig. 1 shows the GC-MS chromatogram of extract. Major phytoconstituents identified are 9-octadecenamide, n-hexadecanoic acid, ethyl linolate, 9-octadecenoic acid (z), 2, 3-dihydroxy-propyl ester, ergost-5-en-3-ol, (3. beta., 24r), gamma-sitosterol, lupeol, fumaric acid, etc. have larger percentage area. Other components with lesser area percentage are butane, 1, 1-diethoxy-3-methyl, 4- isopropylcyclo-hexanone, hexadecane, tetradecanoic acid 1, 2-benzene dicarboxylic acid, and components with least percent area are methoxymethyl trimethyl-silane, methyl stearate, palmitic acid monoethanolamide, alpha.-amyrin. etc.
DISCUSSION: Presence of a large number of therapeutically active components explains the pharmacological potential of F. esceulentum. Majority of the phytoconstituents (hexadecanoic acid, dodecanoic acid, ethoxy carbonyl syringic acid, stigmasterol, heptadecanoic acid, etc.) are antioxidant and thus makes the herb a brilliant antioxidant to detoxify the body. F. esceulentum also possess anticancer potential as it posses antitumor agents like 1, 2 cyclopentanedione, 2-pyrrolidinone 1 methyl, octadecanoic acid, stigma-sterol, alpha-amyrin, etc. Herb also has antihyperglycaemic / anti-diabetic potential which is attributed to 2-ethoxy carbonyl syringic acid, gamma sitosterol, lupeol, stigmasterol, etc. In addition to above components various anti-bacterial, antifungal and anti-insect molecules like 2, 4-Dihydroxy-2, 5-dimethyl-3(2H)-furan-3-one, 4H-Pyran-4-one, 2, 3-dihydro-3, 5-dihydroxy-6-methyl-Hexadecane, hexadecanoic acid, 2-hydroxy -1-(hydroxymethyl)ethyl ester, ethyl linoleate, 1, 2-Benzenedicarboxylic acid, docosanoic acid, ethyl ester are also present.
CONCLUSION: To the best of our knowledge this is the first time when GC-MS is used to determine the chemical composition of F. esculentum extract. Further, every component of the extract can be easily separated and used for deriving new medicinal compounds.
ACKNOWLEDGEMENT: The authors would like to thank Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science & Technology, Hisar, Haryana for providing us the required facilities for the research work.
CONFLICT OF INTEREST: Authors do not have any conflict of interest.
- Li S and Zhang QH: Advances in the development of functional foods from buckwheat. Critical Reviews in Food Science and Nutrition 2001; 41: 451-64.
- Gonçalves FA, Debiage RR, Silva RM, Porto PP, Yoshihara E and de Mello Peixoto EC: Fagopyrum esculentum Moench: A crop with many purposes in agriculture and human nutrition. African Journal of Agricultural Research 2016; 11(12): 983-9.
- Krkošková B and Mrázová Z: Prophylactic components of buckwheat. Food Research International Journal 2005; 38(5): 561-8.
- Gulpinar A, Orhan I, Kan A, Senol F, Celik S and Kartal M: Estimation of in-vitro neuroprotective properties and quantification of rutin and fatty acids in buckwheat (Fagopyrum esculentum Moench) cultivated in Turkey. Food Research International Journal 2012; 46: 536-43.
- Sharma S, Ali A, Ali J, Sahni JK and Baboota S: Rutin: therapeutic potential and recent advances in drug delivery. Expert Opinion on Investigational Drugs 2013; 22(8): 1063-79.
- Kawa JM, Taylor CG and Przybylski R: Buckwheat concentrate reduces serum glucose in streptozotocin-diabetic rats. Journal of Agricultural and Food Chemistry 2003; 51(25): 7287-91.
- Awasthi R: Buckwheat ( esculentum): A gluten free product. Indian Journal of Nutrition 2015; 2(1): 1-5.
- Prakash S and Yadav K: Buckwheat ( esculentum) as a functional food : A Nutraceutical. Pharmacobiology and Medical Sciences 2016; 1(3): 1-15.
- Skoog DA, Holler FJ and Crouch SR: Principles of instrumental analysis. Brooks / Cole Cengage Learning, Edition 6, Vol. I, 2007: 11, 20, 26, 27.
- Hussain SZ and Maqbool K: GC-MS: Principle, technique and its application in food science. International Journal of Current Research 2014; 13: 116-26.
- Nair SC, Kurumboor SK and Hasegawa JH: Saffron chemoprevention in biology and medicine: A review. Cancer Biotherapy and Radiopharmaceuticals 1995; 10: 257-64.
- Bharat CR and Krishna GD: GC-MS analysis of young leaves of Allophylus cobbe (L.) Raeusch and Allophylus serratus (Roxb.) Kurz. Indian Journal of Pharmaceutical Education and Research 2017; 51(3): 472-79.
- Mayr CM, Capone DL, Pardon KH, Black CA, Pomeroy D and Francis IL: Quantitative analysis by GC-MS/MS of 18 aroma compounds related to oxidative off-flavor in wines. Journal of Agricultural and Food Chemistry 2015; 63(13): 3394-01.
- Morita H and Abe I: Plant type III PKS. Hongo, Tokyo: The University of Tokyo 2010: 171-25.
- Janeš D, Kantar D, Kreft S and Prosen H: Identification of buckwheat ( esculentum Moench) aroma compounds with GC-MS. Food Chemistry 2009; 112(1): 120-4.
- Anbukkarasi M, Thomas PA, Sundararajan M and Geraldine P: Gas Chromatography-Mass Spectrometry analysis and in-vitro antioxidant activity of the ethanolic extract of the leaves of Tabernaemontana divaricata. Pharmacognosy Journal 2016; 8(5): 451-58.
- Shriver and Atkins: Inorganic Chemistry. W. H. Freeman and Company, New York, Edition 5, 2010: 358.
- Kumar PP, Kumaravel S and Lalitha C: Screening of anti-oxidant activity, total phenolics and GC-MS study of Vitex negundo. African Journal of Biochemistry Research 2010; 4: 191-5.
- Karthikeyan K, Dhanapal CK and Gopalakrishnan G: GC-MS analysis of ethyl acetate extract of monilifer - Whole plant. Der Pharmacia Lettre 2016; 8(3): 94-99.
- Yang JY and Lee HS: Changes in acaricidal potency by introducing functional radicals and an acaricidal constituent isolated from Schizonepeta tenuifolia. J of Agricultural and Food Chemistry 2013; 61(47): 11511-6.
- Banakar P and Jayaraj M: GC-MS analysis of bioactive compounds from ethanolic leaf extract of Waltheria indica and their pharmacological activities. International Journal of Pharmaceutical Sciences and Research 2018; 9(5): 2005-10.
- Gnanasundaram I and Balakrishnan K: Characterization of bioactive compounds in ethanolic extract of Cissus vitiginea leaves using GC-MS Technique. Journal of Applied Chemistry 2017; 10(9): 24-7.
- Koech DK: Clinical aplications of trioxolane derivatives. African Journal of Health Sciences 2008; 15: 1-2.
- Latimer J, Munday JL, Buzza KM, Forbes S, Sreenivasan PK and McBain AJ: Antibacterial and antibiofilm activity of mouthrinses containing cetylpyridinium chloride and sodium fluoride. BMC Microbiology 2015; 15: 169.
- Nabavi SF, Maggi F, Daglia M, Habtemariam S, Rastrelli L and Nabavi SM: Pharmacological Effects of Capparis spinosa Phytotherapy Research 2016; 30(11): 1733-44.
- Rajapriya S, Geetha A and Ganesan Kripa K: A study on the GC-MS analysis of bioactive components and pancreato-protective effect of methanolic extract of Brassica oleracea var. Botrytis. Natural Product Research 2017; 31: 2174-7.
- Wei A and Shibamoto T: Antioxidant / lipoxygenase inhibitory activities and chemical compositions of selected essential oils. Journal of Agricultural and Food Chemistry 2010; 58(12): 7218-25.
- Das S, Vasudeva N and Sharma S: Chemical compositions of ethanol extract of Macrotyloma uniflorum (Lam.) Verdc. using GC-MS spectroscopy. Organic and Medicinal Chemistry Letters 2014; 4: 13
- Nair RR and Gangaprasad A: GC-MS Analysis of methanolic stem extract of Gynochthodes ridsdalei, Razafim and B. Bremer, an endemic, endangered medicinal plant of southern Western Ghats. International J of Current Pharmaceutical Research 2017; 9(3): 98-01.
- Kumar P, Kumaravel S and Lalitha C: Screening of antioxidant activity, total phenolics and GC-MS study of Vitex negundo. African Journal of Biochemistry Research 2010; 4: 191-5.
- Kalaisezhiyen P and Sasikumar V: GC-MS evaluation of chemical constituents from methanolic leaf extract of Kedrostis foetidissima (Jacq.) Cogn. Asian Journal of Pharmaceutical and Clinical Research 2012; 5(4): 77-81.
- Tyagi T and Agarwal M: Phytochemical screening and GC-MS analysis of bioactive constituents in the ethanolic extract of Pistia stratiotes and Eichhornia crassipes (Mart.) Solms. Journal of Pharmacognosy and Phyto-chemistry 2017; 6: 195.
- Sasikala K and Mohan CS: Total phenolic, flavanoid contents and GC-MS analysis of coromandelicum leaves extract. International Journal of Pharmacy and Pharmaceutical Sciences 2014; 6(8): 379-81.
- Upadhyay RK: Kareel plant: A natural source of medicines and nutrients. International Journal of Green Pharmacy 2011; 5(4): 255-65.
- Gylling H, Plat J, Turley S, Ginsberg HN, Ellegård L and Jessup W: Plant sterols and plant stanols in the management of dyslipidaemia and prevention of cardio-vascular disease. Atherosclerosis 2014; 232: 346-60.
- Misawa E, Tanaka M, Nomaguchi K, Yamada M, Toida T and Takase M: Administration of phytosterols isolated from Aloe vera gel reduces visceral fat mass and improve hyperglycemia in zucker diabetic fatty (ZDF) rats. Obesity Research and Clinical Practice 2008; 2: 239-45.
- Lee CC, Lee BH and Lai YJ: Anti-oxidation and antiglycation of tataricum ethanol extract. Journal of Food Science and Technology 2013; 52(2): 1110-6.
How to cite this article:
Neeraj, Vasudeva N and Sharma S: Chemical composition of Fagopyrum esceulentum Moench seed through GC-MS. Int J Pharm Sci & Res 2019; 10(5): 2392-96. doi: 10.13040/IJPSR.0975-8232.10(5).2392-96.
All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
Neeraj *, N. Vasudeva and S. Sharma
Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science and Technology, Hisar, Haryana, India.
01 September 2018
14 October 2018
18 October 2018
01 May 2019