Metabolic characterization of Achillea millefolium L. through ultraviolet absorption, fourier transform infrared spectroscopy, and gas chromatography–mass spectrometry analysis

Dolly Kain; Suresh Kumar; Jamil Ahmed Khan; Vandana; Amrita Suryavanshi; Atul Arya

doi:10.4103/jdras.jdras_17_21

ORIGINAL ARTICLE

Year : 2021 | Volume : 6 | Issue : 2 | Page : 72-78

Metabolic characterization of Achillea millefolium L. through ultraviolet absorption, fourier transform infrared spectroscopy, and gas chromatography–mass spectrometry analysis

Dolly Kain¹, Suresh Kumar¹, Jamil Ahmed Khan², Vandana³, Amrita Suryavanshi¹, Atul Arya¹
¹ Department of Botany, Medicinal Plant Research Laboratory, Ramjas College, University of Delhi, New Delhi, India
² GMS, Dhararian, Mendhar, Jammu and Kashmir, India
³ Department of Chemistry, Dyal Singh College, University of Delhi, New Delhi, India

Date of Submission	22-Jul-2021
Date of Acceptance	06-Sep-2021
Date of Web Publication	19-Jan-2022

Correspondence Address:
Prof. Suresh Kumar
Department of Botany, Medicinal Plant Research Laboratory, Ramjas College, University of Delhi, Delhi 110007.
India

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jdras.jdras_17_21

Abstract

BACKGROUND: Achillea millefolium L. is an aromatic herbal plant of family Asteraceae having characteristically finely divided leaves that give a fern-like appearance and corymbose cluster inflorescence that gives a flat-headed appearance to the plant. OBJECTIVES: To study absorption patten of mature and immature plant extract using UV spectroscopy, to study the different functional groups present in mature plant extract using Fourier transform infrared spectroscopy (FT-IR), and to study various compounds present in the mature plant extract using gas chromatography mass spectrometry (GC-MS). MATERIALS AND METHODS: Immature and mature A. millefolium have been collected from Pathanteer, Mendhar, Poonch, Jammu and Kashmir. For metabolic characterization of A. millefolium, ultraviolet (UV) absorption, FT-IR, and GC-MS have been used. RESULTS: UV spectroscopy of immature and mature plants reveals the clear difference in their absorption pattern with high content of phenolic and flavonoid compounds in the mature plant. FT-IR analysis of the mature plants reveals the presence of different functional groups including alkyl-substituted ether, secondary amine, and carboxylic acid. GC-MS analysis reveals the presence of 70 compounds including 2-methoxy-4-vinylphenol, phenol, 2,6-dimethoxy, neophytadiene, phytol, ethyl oleate, vitamin E, stigmasterol, γ-sitosterol, and α-amyrin, which are reported to have different medicinal properties. CONCLUSION: Achillea millefolium has shown versatile metabolic composition, which contributes to its different medicinal activities in the literature.

Keywords: Achillea millefolium L., active compounds, FT-IR, functional groups, GC-MS, UV absorption

How to cite this article:
Kain D, Kumar S, Khan JA, Vandana &, Suryavanshi A, Arya A. Metabolic characterization of Achillea millefolium L. through ultraviolet absorption, fourier transform infrared spectroscopy, and gas chromatography–mass spectrometry analysis. J Drug Res Ayurvedic Sci 2021;6:72-8

How to cite this URL:
Kain D, Kumar S, Khan JA, Vandana &, Suryavanshi A, Arya A. Metabolic characterization of Achillea millefolium L. through ultraviolet absorption, fourier transform infrared spectroscopy, and gas chromatography–mass spectrometry analysis. J Drug Res Ayurvedic Sci [serial online] 2021 [cited 2023 Sep 22];6:72-8. Available from: http://www.jdrasccras.com/text.asp?2021/6/2/72/336030

Introduction

Since ancient times, human beings are deriving benefits from nature to improve their well-being. Nature has given almost everything to humans for sustaining life on earth and plants are one of the supreme donations of nature to humans. Plants are not only limited to human welfare but also almost all creatures depend on plants in some or other way. Plants hold an eminent place when it comes to their role in the medical province. Plants from ancient times proved to play a vital role in the treatment of various life-threatening diseases. Traditionally, the plants and their derivatives have been used to cure different ailments in various forms including decoction, juices, infusion, steam inhalation, and powdered form. Today, different extraction methods are available to extract the active portion of the plant. Different separation techniques are available to achieve the isolation of the pure bioactive compounds. Different plants and their isolated bioactive compounds have been successfully used in the treatment of various diseases such as atropine from Atropa belladonna,^[1] vincristine and vinblastine from Catharanthus roseus,^[2] artemisinin from Artimisia annua,^[3] and digitoxin from Digitalis purpurea.^[4]

Achillea millefolium L. belongs to the family Asteraceae, order Asterales, and clade Asterids of kingdom Plantae. It is a perennial herb having a very strong aroma with numerous white flowers in corymbose clusters reported to have different medicinal activities in the literature. It is widely distributed in Central America, North America, Europe, and Asia. In India, it is distributed in the Himalayan region of Himachal Pradesh, Uttaranchal, and Jammu and Kashmir including foothills of Pir Panjal ranges. According to International Union for Conservation of Nature (IUCN), it is listed as species of least concern.^[5] It is mainly considered as a wild species. Genus Achillea was reported to have versatile chemical constituents including terpenoids (monoterpenes, sesquiterpenes, diterpenes, and triterpenes), lignans, flavonoids (cynaroside and cosmosiin), and many other compounds like achillicin, the first natural proazulene identified from the genus Achillea.^[6]Achillea millefolium L. constitutes different phytochemicals such as tocopherols, ascorbic acid, apigenin, quercitin, and caffeoylquinic acid, which were reported to have antioxidant activity; and germacrene D sabinene, borneol, camphor, α-pinene, artemisia ketone, chamazulene, γ-cadinene, and camphene, which were reported to have antibacterial activity. Apigenin and luteolin were reported to have estrogenic activity,^[7] β-pinene, 1,8-cineole, chamazulene, camphor, borneol, and trans-nerolidol have antibacterial activity.^[8] Tocopherols, ascorbic acid, apigenin, quercitin, and caffeoylquinic acid were reported to have antioxidant activity and considered as reducing agents, hydrogen donators, or singlet oxygen quenchers.^[9] Bradykinin and prostaglandins were reported to have diuretic activity.^[10] Luteolin, apigenin-7-0-β-d-glucoside, luteolin 7-0-β-d-glucoside, and 6-OH-luteolin 7-0-β-d-glucoside were reported to have anticholinesterase activity.^[11] Germacrene D sabinene, borneol, camphor, α-pinene, artemisia ketone, chamazulene, γ-cadinene, and camphene were reported to have allelopathic activity, antibacterial, antifungal activity, and anticholinesterase activity.^[12] Apigenin 7-glucoside and luteolin 7-glucoside were reported to have antiplasmodial activity.^[13] Borneol was reported to have antimicrobial activity.^[14]

Materials and Methods

Collection and identification of Achillea millefolium L.

Immature and mature A. millefolium were collected from Pathanteer, Mendhar, Poonch, Jammu and Kashmir, India with GPS location 33º39′40′′N to 74º11′11′′E and identified by Raw Materials Herbarium and Museum (RHMD) and National Institute of Science Communication and Information Resources (NISCAIR), Pusa with reference IDNISCAIR/RHMD/consult/2018/3293–94.

Preparation of extract of Achillea millefolium L.

Stem and leaves of immature and mature A. millefolium were washed with distilled water to remove all the dirt. The washed plant material was then dried under shaded area and ground into fine powder using a mixer grinder for extraction. Stem and leaf extract of immature and mature A. millefolium was prepared in ethanol using the Soxhlet method for 48 h.

Ultraviolet spectroscopy of Achillea millefolium L.

Ultraviolet (UV) absorption spectrum of immature and mature A. millefolium obtained using a spectrophotometer (SP-UV1000) in the range of 200–300 nm.

Fourier transform infrared spectroscopy of Achillea millefolium L.

Fourier transform infrared (FT-IR) spectrum of mature A. millefolium obtained using spectrometer (Shimadzu) in the range of 400–4000 cm⁻¹.

Gas chromatography–mass spectrometry analysis of Achillea millefolium L.

Gas chromatography-mass spectrometry (GC-MS) of mature A. millefolium was performed by using GC-MS program GCMS_QP2010 Ultra with following working conditions: column oven temperature 60°C, injection temperature 260°C, injection mode split, flow control mode linear velocity, pressure 73.3 kPa, flame thermionic detector, ion sources temperature 230°C, and carrier gas saver off.

Results

Ultraviolet absorption of Achillea millefolium L.

UV absorption reveals the clear difference between immature and mature A. millefolium. Immature A. millefolium shows nine peaks at 265, 234, 229, 225, 221, 214, 212, 208, and 203 nm with absorption values of 0.432454, 0.532042, 0.54456, 0.559505, 0.283157, 0.562169, 0.223197, 0.199632, and 0.505235, respectively [Figure 1]A. Mature A. millefolium shows increasing absorption values from 284 nm to 200 nm [Figure 1]B.

Figure 1: Ultraviolet (UV) absorption spectrum of immature (A) and mature (B) Achillea millefolium L.

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Fourier transform infrared spectroscopy analysis of Achillea millefolium L.

Different functional groups have been identified in mature A. millefolium including alkyl-substituted ether, secondary amine, carboxylic acid methyl C–H symmetrical, methyl C–H asymmetrical, and hydroxyl group [Figure 2] and [Table 1].

Figure 2: Fourier transform infrared (FT-IR) spectrum of mature Achillea millefolium L.

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Table 1: Fourier transform infrared (FT-IR) analysis of mature Achillea millefolium L.

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Gas chromatography–mass spectrometry of Achillea millefolium L.

A total of 70 compounds were identified through GC-MS with some major compounds by percentage including 2-methoxy-4-vinylphenol (2.47%), phenol, 2,6-dimethoxy (2.85%), neophytadiene (2.58%), phytol (7.00%), ethyl oleate (1.44%), vitamin E (1.25%), stigmasterol (3.34%), γ-sitosterol (6.27%), and α-amyrin (3.89%) [Figure 3] and [Table 2].

Figure 3: Gas chromatography–mass spectrometry (GC-MS) graph of mature Achillea millefolium L.

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Table 2: Gas chromatography–mass spectrometry (GC-MS) analysis of mature Achillea millefolium L.

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Discussion

Achillea millefolium L. has shown different UV absorption patterns indicating the presence of distinct active chromophores with high phenolic and flavonoid compounds in the mature plant. Almost all phenolic compounds absorb in this UV region; majority of flavonoids absorb in the range between 240 nm and 290 nm and some flavan-3-ol like catechin absorb at 202 nm.^[36] FT-IR reveals the presence of major functional groups of mature A. millefolium. Different functional groups provide different medicinal properties to the biological compounds.^[37] GC-MS reveals the presence of 70 active compounds of mature A. millefolium. Achillea millefolium L. has shown versatile metabolic composition with many active compounds responsible for its different medicinal activities. The predominant constituents of A. millefolium essential oil include sabinene, 1,8-cineole, borneol, bornyl acetate, pinene, $-pinene, terpinine-4-ol, and chamazulene.^[38]

Clinical significance

This study reveals the potential metabolites of A. millefolium responsible for various medicinal activities and provided the basis for isolation and characterization of the main bioactive compounds having specific activity. It is reported to have different medicinal properties including anti-inflammatory, blood purifier, hepatoprotective, diuretic, antispasmodic, estrogenic activity allelopathic activity, antibacterial, antifungal activity, anticholinesterase activity, and antimicrobial.

Acknowledgement

The authors thank the Principal, Dr. Manoj K. Khanna, Ramjas College, Prof. S. B. Babbar, Prof. K.S. Rao, and Prof. Veena Agrawal, Department of Botany, University of Delhi, Delhi for providing necessary facilities and encouragement during the investigation. The authors are acknowledging the experts to review the manuscript for improvement. The authors are also thankful to University Grant Commission (UGC) for the financial support.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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