Identification Lipid compounds of Aquatic plant Hydrilla verticillata (Linn. F.) Royal

 

Sadiq Kadhum Lafta Alzurfi1, Mustafa Abd Alkadhum Rabeea2,

Jaafar B. Algburi3, Nour Najah Ibrahimi4,

Mohammed Jawad Salih Al-Haidarey5

1,3,4,5Dep. of Ecology, Uni. of Kufa-Iraq

2Dep. of Chemistry Uni. of Kufa-Iraq

*Corresponding Author E-mail: mohammedj.alhayderi@uokufa.edu.iq

 

ABSTRACT:

Introduction: Gynecological disorder is a condition that affects female reproductive system, mainly breasts and organs in the pelvic area such as uterus, ovaries, fallopian tubes, vagina, and vulva. Hence, the term encompasses a broad term of diseases that can affect female fertility such as pelvic inflammatory diseases and endometriosis. Hydrilla verticillata (Linn. F.) considered as an invasive species in Iraq. Our study aimed to identified the concentration of lipid compounds in that plant whicht collected from Euphrates River/Kufa-Iraq. Twenty-three compounds were identified. The most prevailing compounds were Hexadecanoic acid, methyl ester (26.26%), 9,12-Octadecadienoic acid (Z, Z)-, methyl ester (19.07%) and phytol (11.83%) as major components, and their structures were elucidated. In present study, we concluded that this species (H. verticillata) is a good source of many fatty acids especially octadecadienoic acid and phytol, and we suggested to create new studies deals with using this plant in pharmaceutical products.

 

KEYWORDS: Euphrates, GC-MS, Hydrilla verticillata, lipid, Phytol.

 

 

 

INTRODUCTION:

Hydrilla verticillata, is a submerged freshwater angiosperm plant, distributed across Africa, Eurasia, America, and Oceania (1,2). This species was introduced in the aquatic ecosystems of Florida waters during 1960 and distributed to all of drainage basins in all US by the early 1970s (3). Now, we found it in all continent except Antarctica (1).

 

Phytochemical analysis revealed the presence of alkaloids, phenols, flavonoids, saponins and terpenoids. Gas Chromatography-Mass Spectrometry (GC-MS) study of the ethanolic extract of Hydrilla verticillata confirmed the presence of phytol and hexadecanoic acid ethyl ester as major components (4). Since it contains rich minerals and vitamins, it is called “green food”.

 

Hydrilla is enriched with vitamin B (especially vitamin B12), calcium, fatty acid (like gamma linoleic acid), antioxidants (such as zinc), selenium, SOD, vitamin C, and proteins (5), Xiao et, al (6) mention to diagnosis five compounds which isolated by column chromatography and purified from the extractions of Hydrilla verticillata (Linn. f.) Royle includes: 1-(5′-Hydroxy-4′-hydroxymethyl-1′-methyl-1H-pyrrol-2′-yl)-henicosa-2,12,15-trien-1-one (2) and thymidine (3) were identified by EIMS, 1H NMR, 13C NMR and IR spectra. Crystal loliolide (1) and sulfur-gamma (4) were identified by X-ray diffraction. Octadecanedioic acid (5) was also elucidated. Among them, compound 2 is a new natural compound, loliolide (1) and thymidine (3) are known bioactive substances. Byju et al, (2013), during GC-MS study of the unsaponifiable lipophilic extract of H. verticillata, find 3, 5, 11, 15-tetramethyl 1- hexadecen-3-ol (C20H40O) and phytol (C20H40O) as major components, thus the phytol and 3, 5, 11, 15-tetramethyl 1- hexadecen3-ol are the two isomers of the diterpeneol (C20H40O) found in the unsaponifiable lipophilic and are formed by the hydrolysis of the alcohol moiety of chlorophyll. Since there is no study dels with the lipids in that plant, this study comes to identification lipids compounds in H. verticillata.

 

MATERIAL AND METHODS:

Hydrilla verticillata was collected during August 2016 from Euphrates River in Zerga region which located 5 km north Kufa City (29 32̊ North, 16 44 ̊ East), the plant washed carefully in the field by river water and in the lab by deionized water several times to remove of algae and small animal. Sixty gram of the dry grained sample was extracted with methanol: benzene (1:1) ratio, two hundred milliliter of the methanol: benzene mixture was collected and the extract was concentrated in vacuum and the crude residue was collected. The fatty acid compounds were separated by saponification according to (7,8). Then, fatty acids were identified by using GC -MS spectroscopy (model 2010Qp Plus shimadzu) in the conditions of 40.0°C  oven temperature, 280°C injection temp, the separation was done within 30 min, the gain was 0.70 kV +0.10 kV, the inlet line and the source temperatures were set at 200°C. Fourier Transform Infrared Spectrophotometer (FTIR) is perhaps the most powerful tool for identifying the types of chemical bonds/functional groups present in the phytochemicals. Dried powder of methanol solvent extracts of H. verticillata was used for FTIR analysis. The powdered sample of each extract was loaded in FTIR spectroscope (bruker, Germany), with a Scan range from 400 to 4000 cm1. 

 

RESULTS AND DISCUSSION:

FTIR analysis.

The results of FTIR analysis for Hydrilla verticillata plant extraction (Fig 1) showed that there is a clear and high intensive band (3308/cm) which represent OH groups of methanol extract and phenol group of Phytol. The moderate band (2921-2850/cm) that represents frequency a symmetrical patterns of group CH2 and CH3 of alcoholic compound. Another band (1733/cm) represented frequency patterns of ester group (Hexadecanoic acid, methyl ester). Which belong to compounds containing carbonyl groups C = O, which are either (9, 12, 15-Octadecatrienoic acid, methyl ester) the band 1643/cm which is also the patterns match the frequency of the carbonyl group belonging to quinine (9) which nearby and similar to aromatic group (C=C) of solvent and aliphatic compounds of phytol. The bands at 1456/cm - 1418 /cm represents the frequency of pattern of group CH2- and other strong band (1043 /cm) which represents frequency of pattern of group C-O, indicates the presence of carboxylic acids found in the compound known (chromopropanecarboxylic acid).

 

GC-MS Analysis:

The Hydrilla verticillata extract was analyzed by Gas Chromatography-Mass spectrometry at the optimum separation conditions described by Iordache, et al (10). The results showed several distinct peaks as shown in fig 2. The peak at 21.581min time has a mass spectrum at a base peak at M / Z = 74 attributable to Hexadecanoic acid, methyl ester as shown in fig 3, evidence on literature has shown that Hexadecanoic acid isolated from a marine red alga may be a lead compound of anticancer drugs (11). The second characteristic peak is at the retention time 23.118 min gives a peak at M / Z = 67 due to the 9, 12-Octadecadienoic acid, methyl ester compound as in fig 4, possessed with anti-inflammatory and cancer preventive characters (12).

 

While the peak at a retention time (23.168 min) has a peak mass (M / Z = 79) due to a 9, 12, 15-Octadecatrienoic acid, methyl ester compound, as shown in Figure (5). These essential fatty acids belonging to the C18 fatty acid differ structurally in the position and degree of unsaturation (13). Recently these essential fatty acids are used as health supplements because of the health benefits associated with (14). That the apparent peak at the retention time (23.250 min) was identified as a compound with a peak mass of (M / Z = 71) due to Phytol as shown in Figure (6), (15, 16). The chlorophyll is most abundant photosynthetic pigment in higher plants. The presence of the phytol, diterpene alcohol can be considered as common among plant volatiles, knowing that it is a hydrolysis product of the alcohol moiety of chlorophyll (17). Another class of phytyl esters was also detected in plants, i.e. fatty acid phytyl esters (18). In H. verticillata also, the E-isomer is predicted by the library check. Both Z and the E phytols have been isolated as unbounded molecules from the red algae, Gracilaria andersonia (19). The current study founded more lipids compounds in H. verticillata than Byju et al ,(20) study that recorded fifteen compounds while Pandi and Rajkumar(4) found six compounds.

 

Figure (2) Chromatogram Gas Chromatograph - Mass spectrometry

 

Figure (3) Mass fragmentation of (a) Methyl tetradecanoate (b) Tetradecanoic acid, methyl ester (C) Myristic acid, methyl ester d) Metholeneat e) Methyl myristate f) Methyl n-tetradecanoate extract of Hydrilla verticillata separated by GC.

 

Figure (4) Mass fragmentation of (a) 9,12-Octadecadienoic acid (Z,Z)-, methyl ester (b) Linoleic acid, methyl ester (C) Methyl cis,cis-9,12-octadecadienoate d) Methyl linoleate e) Methyl octadecadienoate f) Methy l 9-cis,12-cis-octadecadienoate g) Methyl (9Z,12Z)-9,12-octadecadienoate extract of Hydrilla verticillata separated by GC.

Figure (5) Mass fragmentation of (a) 9,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z)-(b) Linolenic acid, methyl ester (C) Methyl all-cis-9,12,15-octadecatrienoate d) Methyl linoleate e) Methyl (9Z,12Z,15Z)-9,12,15-octadecatrienoate extract of Hydrilla verticillata separated by GC

 

Figure (6) Mass fragmentation of (a) Phytol (b) 2-Hexadecen-1-ol, 3,7,11,15-tetramethyl-, [R-[R*,R*-(E)]]- (C) trans-Phytol d) 3,7,11,15-Tetramethyl-2-hexadecen-1-ol E) (2E)-3,7,11,15-Tetramethyl-2-hexadecen-1-ol extract of Hydrilla verticillata separated by GC.

 

The current study concluded that Hydrilla verticillata has 23 fatty acid compounds in Hexadecanoic acid and methyl ester forms. Octadecadienoic acid and phytol were found in the highest amount. Phytol compound in the Hydrilla verticillata was sufficient amount.

 

ACKNOWLEDGEMENT:

We would like to thank Dep. of Ecology, University of Kufa for funding this study.

 

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Received on 15.04.2019           Modified on 23.05.2019

Accepted on 18.06.2019         © RJPT All right reserved

Research J. Pharm. and Tech. 2019; 12(11):5621-5626.

DOI: 10.5958/0974-360X.2019.00973.9