GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS OF HUGONIA MYSTAX LEAVESHTML Full Text
GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS OF HUGONIA MYSTAX LEAVES
Vasuki *, P. Chitra, M. Vijayabaskaran, N. Mahadevan and R. Sambathkumar
Department of Pharmaceutical Chemistry, J. K. K. Nattraja College of Pharmacy, Kumarapalayam, Tamil Nadu, India.
ABSTRACT: Hugonia mystax is a woody evergreen plant that grows in the dry forest areas of India and Sri Lanka. Often found in the forests of Kerala and Tamil Nadu in southern India. It is locally known as modirakanni (tamil) and kamsamaraha and tribals from Tamil Nadu, India, have used it in primary health care. The plant parts are used ethnobotanically to treat rheumatism, skin diseases and inflammations, snake bites, fever, and worm infestation. Hugonia mystax is an antibiotic, anthelmintic, febrifuge, astringent, peptic ulcer and verminosis remedy. Hugonia mystax has been practising herbal medicine for a long time. This plant is a member of the linaceae family and is an essential medicinal plant in Indian medicine. To evaluate the extractive values, powdered materials were subjected to successive extractions with ethanol using the Soxhlet process. The existence of several compounds such as flavonoids, phenolic elements, steroids, tannins, saponins, terpenoids, and carbohydrates was revealed by qualitative phytochemical screening of the ethanolic extracts of the leaves, as well as GC-MS analysis to examine the chemical components present in it. A total of twenty-seven compounds were discovered in the ethanolic extract. The findings of this study would form the basis for the manufacture of herbal medicines for a variety of ailments using Hugonia mystax leaves.
Keywords: Hugonia mystax, Phytochemical analysis, Soxhlet extraction, GC-MS
INTRODUCTION: In India there are about 7500 wild plants used for medicinal purposes by different tribal inhabitants. Treatment with medicinal plants were considered very safe as there is no or minimal side effects. The golden fact is that, use of herbal treatment is independent of any age group 1. Most of the drugs thus formulated are free of side effects or reactions. One of the phenomena of the last three decades has been the huge increase in use of herbal products 2.
These can be defined as plants, parts of plants or extracts from plants that are used in health care or in combating diseases 3. Traditional plant-based medicines for primary healthcare used is followed in underdeveloped countries of about 80% of world’s population.
Hugonia mystax leaves were associated with different biological activities such as anti-microbial, anthelmintic, anti-oxidant, hepato-protective and cytotoxic actions. This species has a restricted global distribution, occurring only in India and Sri lanka. Within India, it has been recorded in Maharashtra, Karnataka, Tamil nadu and Andhra Pradesh. It is an unexplored medicinal plant in the Indian medicinal system. Medicinal plants are the richest bio resources of drugs of traditional systems of medicine, modern medicines, neutraceuticals, food supplements, folk medicines, pharmaceutical intermediates and chemical entities for synthetic drugs 4. The plant kingdom holds a great promise for medicinal substances in many plant species and still unexplored 5.
According to ethnobotanical information, the leaves are used in the treatment of peptic ulcers and as anthelmintic, febrifuge, antidote and its fruits are used in diarrhea and dysentery 6. Stem bark is used in the treatment of jaundice and skin diseases 7.
All these medicinal properties of Hugonia mystax is due to the presence of its phytochemical constituents which is not yet explored thoroughly 8. In this study the gas chromatogram mass spectrometric method was carried out in the ethanolic extract of dried leaves for phytochemical analysis 9.
There are thousands of phytochemicals each have their pharmacological properties of their own. The plant possesses various medicinal properties. The aim of this study was to identify the phyto-compounds in the ethanolic leaf extract of the Hugonia mystax by qualitative screening of phytochemicals and to identify each specific compound by GC-MS analysis. A major part of the world population mainly in the developing countries still uses traditional, folk medicine to avoid synthetic drugs treatment.
FIG. 1: HUGONIA MYSTAX
Recently many of the research were being carried out in medicinal plants. The main reason was that the synthetic drugs which was now taking up by the human have many side effects that often lead to serious complications. The development of herbal medicine was done by the primary screening of the compounds in the plant extracts.
Comparing to modern medicine the herbal medicine was the lifesaving drug 10. The non-nutritive plant chemicals are called as phyto-chemicals which have the properties to protect or prevent diseases. Plant produces these chemicals to protect themselves but the research shown that they have the capacity to treat human diseases in an effective way 11. Review of literature revealed less work on this plant. Hence in the present study, the successive extractive value and Gas chromato-graphy-Mass spectrometry analysis of ethanol extract of leaves of Hugonia mystax were done.
MATERIALS AND METHODS:
Collection of Plant Material: The leaves of Hugonia mystax were collected from Dindigul District, Tamil Nadu, India. The collected plant material was authenticated by Botanical Survey of India (BSI/SRC/5/23/2018/Tech/149). The herbarium specimen was prepared and deposited at the Natural Product Research Laboratory, J.K.K. Nattraja College of Pharmacy, Namakkal, Tamil Nadu for future reference.
Preparation of the Extract: The plant material was collected and chopped into small pieces, dried under shade condition and coarsely powdered 12, 13. The coarse powder was subjected to successive extraction with ethanol solvent by Soxhlet extraction method 14. The extract was collected and distilled at atmospheric pressure and the trace of solvent was removed in vacuo and stored at 4 ºC. The resulted extract was subjected to GC-MS analysis 15. Table 1 shown the preliminary phyto-chemical analysis of dry leaf extract of Hugonia mystax Linn 16.
TABLE 1: PRELIMINARY PHYTOCHEMICAL ANALYSIS OF DRY LEAF EXTRACT OF HUGONIA MYSTAX
|Phytochemical Constituents||Ethanol extract|
|Fats and oils||-|
+ ---- Present, - ---- Absent
TABLE 2: GAS CHROMATOGRAPHY-MASS SPECTROMETRY 17
|Make||Agilent 7890 B with Mass Detector|
|Column||HP-5MS, 30m × 0.250 mm, 0.250μm|
|Collision gas (N2)||1.50 mL/minute|
|Quench gas (He)||2.25 mL/minute|
|Injection Volume||1.0 μL with split|
|Inlet Temperature||200 °C|
|Initial Temperature||90 °C|
|Initial Time||6 minutes|
|Program rate 1||6°C/minute|
|Temperature 1||280 °C|
|Hold Time 1||23 minutes|
|Run time||60.667 Minutes|
|Injection mode||Split 20:1|
|Auxiliary temperature||230 °C|
|Ion source Temperature||230 °C|
|Solvent cut time||3 minutes|
|Detector Gain Mode||Relative to Tuning result|
The details of gas chromatography mass spectrometry are shown in Table 2 18.
Identification of Compounds: The individual compounds were identified from ethanol extract based on direct comparison of the retention times and their mass spectra with the spectra of known compounds stored in the spectral database 19, 20, NIST version 2.2 mass spectral library and comparing the spectrum obtained through GC-MS compounds present in the plant samples were identified 21.
Compounds Identification by GC-MS: The samples eluted using column chromatography has been analysed using gas chromatography equipped with mass spectrometry.
Elution solvent of ethanol was used 22, 23. Hence the blank solvents and separated samples were injected in the GCMS and chromatograms were obtained 24. The result of the sample was tabulated.
RESULTS AND DISCUSSION:
TABLE 3: COMPONENTS DETECTED IN THE LEAF OF ETHANOL EXTRACT OF HUGONIA MYSTAX
FIG. 2: GAS CHROMATOGRAPHY MASS SPECTROMETRY ANALYSIS25
TABLE: 4 BIOLOGICAL PROPERTIES OF THE PHYTOCOMPONENTS
|Name of the compound||Compound nature||Activity|
|Methyl 4-nitrohexanoate||Aliphatic ester||Lubricant|
|Naphthalene||Polycyclic Aromatic hydrocarbon||Anti-microbial|
|2-Methyl-1-ethylpyrrolidine||Cyclic amine||Anti-bacterial activity|
|5-Hydroxy methyl furfural||Aldehyde||Anticancer|
|Ethyl isoxazole-5-carboxylate||Heterocyclic ester||Anti-oxidant, anti-cancer, anti-tubercular|
|Melezitose||Non reducing sugar||Stabilizing human blood plasma proteins|
|Folic Acid||Vitamin compound||Treatment of anemia|
|Acetamide, N-methyl-N-[4-[2-acetoxymethyl-1-pyrrolidyl]-2-butynyl]-||Cyclic ester||Anti-biotic, anti-cancer|
|Z-(13,14-Epoxy) tetradec-11-en-1-ol acetate||Cyclic ester||Anti tumor|
|Cyclopentanone, 2-(2-octenyl)-||Cyclic ketone||Pesticide|
|Isopropylpalmitate||Aliphatic Ester||Emollient, moisturiser|
|Tridecanedioic acid||Aliphatic dicarboxylic Acid||Anti-biotic|
|Trans Sinapyl alcohol||Aromatic Alcohol||Hepatoprotective agent|
|9, 12-octa decadienoic acid||Linoleic acid||Anti-inflammatory, ant histaminic, antiandrogenic, antiarthritic|
|13-heptadecyn-1-ol||Terpenes||Anti-inflammatory, anticancer, antimalaria|
|Hexadecanoic acid, ethyl ester||Palmitic acid ester||Anti-oxidant, lubricant, pesticide, anti-androgenic, antibacterial|
|Oleic acid||Unsaturated aliphatic Acid||Anti-inflammatory, Pulmonaryedema, cancer preventive|
|Estra-1,3,5 (10)-trien-17beta-ol||Alcohol||Act as a human metabolite|
|11beta, 17beta-dihydroxy-19-nortestosterone||Steroid||Anti-cancer and anemia|
|Vitamin E||Vitamin compound||Anti-ageing, anti-diabetic, anti-cancer, hepatoprotective|
|6,7-Epoxypregn-4-ene-9,11,18-triol-3,20-dione,11, 18-diacetate||Steroid||Anti-microbial, anti-cancer, anti-inflammatory|
|8-(2,5-Dimethoxyphenyl)-6-methyl-2-(4-methylpent-3-enyl) octa-2,6-dienoic acid, ethylester||Aromatic acid and ester||Treatment of respiratory diseases|
|Stigmasterol||Steroids||Lowering the LDL, anti hypercholesterolemic, antitumor|
|Beta sitosterol||Steroids||Cardiac disorder, anti-cancer|
The results of GC-MS analysis on an ethanol extract of leaves are shown in Table 3 26. Preliminary phytochemical analysis was carried out to qualitatively analyze the phytoconstituents in the extracts.Twenty-seven compounds were identified with ethanol extract of leaves of Hugoniamystax. All 27 compounds are medicinally important.The presence of various bioactive compounds confirms the application of Hugoniamystax for various ailments by traditional practitioners 27.
Isolation of individual phytochemical constituents may proceed to find a novel drug. In addition to this, the results of the GC-MS profile can be used as phytochemical tool for the identification of the bioactive components. The active principles with their retention time (RT), molecular formula, molecular weight (MW) and concentration (%) in the ethanolic extract of Hugoniamystax are presented in Table 3.
Methyl 4-nitrohexanoate (13.23%), Naphthalene (15.22%), 2-Methyl-1-ethylpyrrolidine (59.91%), 5-Hydroxy methyl furfural (23.19%), Ethyl isoxazole-5-carboxylate (41.15%), 2-Hydroxy-5-methylbenzaldehyde (19.53%),Melezitose (23.96%), (E)-4-(3-Hydroxyprop-1-en-1-yl)-2-methoxyphenol (37.38%), Z-(13, 14-Epoxy) tetradec-11-en-1-ol acetate (13.86%), Isopropyl-palmitate (29.45%), Trans Sinapyl alcohol (13.99%), 13-heptadecyn-1-ol (11.06%), 6,7-Epoxypregn-4-ene-9, 11, 18-triol-3, 20-dione, 11, 18-diacetate(14.13%), 8-(2, 5-Dimethoxy-phenyl)-6-methyl-2-(4-methylpent-3-enyl)octa-2,6-dienoic acid, ethylester (23.73%), Beta sitosterol (18.41%). All these compounds present in Hugoniamystax leaf extract support the medicinal application of the plant.The study revealed major bioactive compounds present in the ethanol extract. Identification of these compounds in the plant serves as the basis in determining the possible health benefits of the plant leading to further biologic and pharmacologic studies.
CONCLUSION: According to the findings of this report, the plant Hugoniamystax leaves, as well as the chemical constituents contained in them, are highly valuable in medicinal use for the treatment of various human ailments.The existence of medicinally valuable bioactive components is revealed by GC-MS study of the ethanolic extract of Hugoniamystax leaves. To develop safe drugs, further research on toxicological aspects is needed, as well as the isolation, purification and identification of the bioactive molecules responsible for the activities in order to develop novel pharmaceutical leads.
ACKNOWLEDGEMENT: The authors wish to express our thanks to J. K. K. Nattraja College of Pharmacy, Kumarapalayam, Namakkal District, Tamilnadu for providing the facility to conduct this research work.
CONFLICTS OF INTEREST: The authors declare no conflicts of interest.
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How to cite this article:
Vasuki B, Chitra P, Vijayabaskaran M, Mahadevan N and Sambathkumar R: Gas chromatography-mass spectrometry analysis of Hugonia mystax leaves. Int J Pharm Sci & Res 2022; 13(1): 409-16. doi: 10.13040/IJPSR.0975-8232.13(1).409-16.
All © 2022 are reserved by the International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
B. Vasuki *, P. Chitra, M. Vijayabaskaran, N. Mahadevan and R. Sambathkumar
Department of Pharmaceutical Chemistry, J. K. K. Nattraja College of Pharmacy, Kumarapalayam, Tamil Nadu, India.
25 April 2020
17 December 2020
20 December 2020
01 January 2022