{"id":54261,"date":"2023-12-31T10:40:12","date_gmt":"2023-12-31T10:40:12","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=54261"},"modified":"2024-01-05T06:53:37","modified_gmt":"2024-01-05T06:53:37","slug":"extraction-isolation-and-structure-elucidation-of-two-phenolic-acids-from-aerial-parts-of-celery-and-coriander","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no4\/extraction-isolation-and-structure-elucidation-of-two-phenolic-acids-from-aerial-parts-of-celery-and-coriander\/","title":{"rendered":"Extraction, Isolation and Structure Elucidation of Two Phenolic Acids from Aerial Parts of Celery and Coriander"},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Several therapeutic compounds have been detected in medicinal herbs\ntoday. These herbs are recorded in pharmacopeia throughout\nthe world and play an important role in conventional medicine1<\/sup>.\nCoriandrum sativum <\/em>L. (known as coriander) is a seasoning plant of the\nApiaceae family, with origins in Mediterranean\ncountries. This plant is grown commercially in Asia, Africa, and Europe2<\/sup>. All parts of the plant\nare edible while the common parts utilized in cooking are the dried seeds and\nthe fresh leaf 3<\/sup>. Coriander (see Figure 1) has nutritional components such as protein, water, fiber, fat, ash,\nsugar, essential oils, and minerals4<\/sup>.\nThe main active ingredients of coriander are\nfatty oils and essential oils5<\/sup>. It also contains coumarins,\nphthalides, phenolic acids (chlorogenic and caffeic acids), and flavonoids\n(quercetin and rutin)6,7<\/sup>. Oleic, linoleic, palmitic, ascorbic, and\nstearic acids are also present in the coriander plant and\nare very effective in lowering the blood cholesterol level8<\/sup>.\nCoriander is used in Indian folk medicine for the treatment of urinary, digestive, and\nrespiratory disease; it also has diuretic,\ndiaphoretic, stimulant, and carminative activity3<\/sup>.\nThis plant exhibits a strong antioxidant\neffect because it is an excellent source of phytochemicals and polyphenols2<\/sup>.\nThe coriander leaf contains higher antioxidant\nconcentrations than its seeds5<\/sup>. It also possesses other pharmacological properties such as anti-mutagenic\n9<\/sup>, antidiabetic 10<\/sup>, anti-spasmodic 11<\/sup>, antilipidemic effects 12<\/sup>, and others.<\/em><\/strong><\/p>\n\n\n\n

Apium graveolens <\/em>L. (known as celery) is an annual plant of the Apiaceae family, native to outlying hills in Afghanistan,\nPunjab, and the North Western Himalayas13<\/sup>.\nThe leaves, roots, and stems are vastly\nutilized in cooking such as salads, side dishes, and soups. Celery contains furanocoumarin, polyphenols, tannins, steroids, terpenoids,\nsaponins, acids (glutamine, ascorbic, oxalic), vitamins B2, B1, A, PP, B9, B6,\nK, E, and mineral salts (14)<\/sup>.  It also contains essential oils as the main active constituents14<\/sup>. It is\napplied in traditional medicine to the\nbladder, kidney diseases, gout, gastric ulcer, gastritis, duodenal gout,\nobesity, diabetes, dermatitis, nephritis, rheumatism, and prostate inflammation14<\/sup>. The celery plant has different activities such as antidepressant,\nantimicrobial, antidiabetic13<\/sup>,\nhepatoprotective, estrogenic, antioxidant,\nantiestrogenic, cytotoxic15-17,<\/sup> anti-inflammatory and\nanalgesic effects18,19<\/sup>.<\/p>\n\n\n\n\n\n
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Figure 1: Photo of Iraqi <\/strong>Coriandrum sativum<\/em><\/strong>(A) and Iraqi Apium graveolens<\/em>(B).<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Materials and Methods<\/strong><\/p>\n\n\n\n

 Apparatus and Instruments <\/strong><\/p>\n\n\n\n

The following were used, Rotatory\nevaporator type Buchi attached to a vacuum pump Buchi-Germany), oven\n(Memmert-854, Buchi-Germany), and an electrically sensitive balance (Sartorius,\nGermany) with an Ultraviolet light type DESAGA HEIDELBERG of 254 nm and 366 nm\nwaves lengths type DESAGA-Germany. Fourier transforms infrared spectra (FT-IR)\nspectra were scanned on Jusco\/ Japan FT-IR-4200 at the University of Baghdad,\nCollege of Pharmacy, 1H NMR was carried out in al-Albayt University, Al-Mafraq,\nJordan (Euro-vector EA 3000A\/ Italy), LC\/MS was carried out in Iraqi National\nCenter for Drug Control and Research (LCMS-8040 series system, Shimadzu\/ Japan)\nand HPLC analysis was carried out using SYKAM\/ Germany at the directorate of the ministry of\nscience and technology\/environment and water center.<\/p>\n\n\n\n

Chemicals and Reagents<\/strong><\/p>\n\n\n\n

Standard of p-coumaric acid and\nCaffeic acid were purchased from Biopurfy-China, Silica gel of GF 254nm<\/sub>\nwith a thickness of 0.25 mm MERCK-USA while the 0.75mm from Sanpont-China,\nabsolute both ethanol and methanol, HPLC grade acetonitrile, the ethyl acetate\nand petroleum ether were purchased from Schar lab S.L.-Spain and Toluene was\npurchased from the CDH-India while the formic acid was from GCC, U.K.<\/p>\n\n\n\n

Plant material<\/strong><\/p>\n\n\n\n

The aerial parts of the plants (Apium graveolens<\/em> L. and Coriandrum sativum<\/em>) were collected from\nBaghdad city, which is located in the center of Iraq, in\nDecember 2021. Some aerial parts of each plant were dried and the other aerial parts\nwere used fresh.<\/p>\n\n\n\n

Plant\ncollection licenses <\/strong><\/p>\n\n\n\n

We have\nreceived ethical approval and permission to collect celery and coriander from the Pharmacognosy\nDepartment at Baghdad University, and Iraqi Medical Research Center 1120,2020. <\/strong><\/p>\n\n\n\n

Extraction of phenolic acids<\/strong><\/p>\n\n\n\n

The extraction\nwas performed according to the process with\nsome modifications described by Iswantini et al<\/em>.<\/em> <\/sup>20,21<\/sup>. The extraction was carried out for dry and\nfresh aerial parts (leaves and stem) of Apium graveolens<\/em> L.\nand Coriandrum sativum <\/em>L., taking (100 gm) of each dry plant and fresh parts\nand then macerated with 70 % ethanol (1000 ml) at room temperature separately.\nThe extracted crudes were concentrated under reduced pressure at 50 \u00b0 C. Then\nsuccessively fractionated with petroleum ether (250 ml*2) and ethyl acetate\n(250 ml*2). The ethyl acetate fractions were concentrated separately to obtain\nfour fractions of ethyl acetate for each part of the plant.<\/p>\n\n\n\n

Identification of phenolic acids by thin-layer chromatography (TLC)<\/strong><\/p>\n\n\n\n

The\npreliminary analysis of caffeic acid (C9<\/sub>H8<\/sub>O4<\/sub>)\nand p-coumaric acid(C9<\/sub>H8<\/sub>O3<\/sub>) in ethyl\nacetate fractions from fresh and dry aerial parts of Apium graveolens <\/em>and\nCoriandrum sativum <\/em>was achieved by TLC.<\/p>\n\n\n\n

The GF254<\/sub> plate of silica gel (readymade) was used as the stationary phase and toluene: ethyl acetate: formic acid\n(36:12:5) as developing solvent 21<\/sup>.\nThe detection of developed plates is done first\nby visualizing under UV light at 254nm, then spraying with a chemical reagent\nfor phenolic compounds such as ferric chloride\n(0.5%). The value of the retardation factor (Rf value)of the separated spots of caffeic and p-coumaric acids was calculated\nand matched to that of their standard.<\/em><\/p>\n\n\n\n

Qualitative and quantitative\nestimations of phenolic acids by high-performance liquid chromatography (HPLC)<\/strong><\/p>\n\n\n\n

By HPLC (table 1), qualitative analysis of caffeic and p-coumaric acids in ethyl acetate fractions of celery and coriander plants is performed by matching\nthe retention time of caffeic and\np-coumaric acids in all samples with those of their standard at\nidentical chromatographic conditions 22<\/sup>: <\/p>\n\n\n\n

Table 1: HPLC conditions<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Mobile phase(isocratic)<\/p>\n<\/td>\n

\n

Acetonitrile: Water (1:1)<\/p>\n<\/td>\n<\/tr>\n

\n

Column<\/p>\n<\/td>\n

\n

Shimadzu LC C18 (250 mm x 4.6 mm, 5 \u03bcm particle size)<\/p>\n<\/td>\n<\/tr>\n

\n

Column temperature<\/p>\n<\/td>\n

\n

ambient<\/p>\n<\/td>\n<\/tr>\n

\n

Flow rate<\/p>\n<\/td>\n

\n

1ml \/ min<\/p>\n<\/td>\n<\/tr>\n

\n

Injection volume<\/p>\n<\/td>\n

\n

20\u03bcL<\/p>\n<\/td>\n<\/tr>\n

\n

Injection concentration<\/p>\n<\/td>\n

\n

1 mg \/ml<\/p>\n<\/td>\n<\/tr>\n

\n

Detection<\/p>\n<\/td>\n

\n

UV detector at \u03bb 265 nm<\/p>\n<\/td>\n<\/tr>\n

\n

Sample<\/p>\n<\/td>\n

\n

Ethyl acetate fraction<\/p>\n<\/td>\n<\/tr>\n

\n

Standards<\/p>\n<\/td>\n

\n

Caffeic acid and p-coumaric acid<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

In HPLC, quantitative estimations of p-coumaric and caffeic\nacids in ethyl acetate fractions of celery and coriander plants are made by drawing a calibration chart for which\nserial dilutions of the standard caffeic and p-coumaric acids were made by\nmethanol (10, 20,30, and 40 ppm). <\/p>\n\n\n\n

Isolation and purification\nof phenolic acids by preparative TLC plates<\/strong><\/p>\n\n\n\n

The isolation of caffeic and p-coumaric acids from the ethyl acetate fraction of the dry coriander aerial part(DC) is achieved by\nusing preparative TLC. TLC plates (silica gel\nGF254<\/sub> with thick 0.75mm and 20*20) were used and activated at 120\u00b0 <\/sup>C before the application of the sample. The ethyl\nacetate fraction of DC is seen by a capillary\ntube on the plate in the form of a row and,\naccording to standard processes, allowed the plate to develop\nin toluene: ethyl acetate: formic acid (36:12:5) as a\nsolvent system.  Under UV at 254nm, the\nseparated bands were identified and with a needle marked. The two bands were\nrubbed off with standards similar to those of the\np-coumaric and caffeic acids standards put in a beaker\nand then adding a sufficient quantity of absolute methanol, the flasks\nwere later shaken on a warm water bath, kept cool in a refrigerator for\ncrystallization about 4-5 hrs. The crystalline constituent was then filtered\nthrough double filter paper and dried for a pure constituent 23<\/sup>.<\/p>\n\n\n\n

Identification and characterization\nof the isolated constituents <\/strong><\/p>\n\n\n\n

Fourier transforms infrared (FT-IR) spectroscopy <\/strong><\/p>\n\n\n\n

The FT-IR spectroscopy was performed in Baghdad University\/College\nof Pharmacy\/ Department of Pharmacognosy and Medicinal plants for each isolated\nconstituent by using a KBr disc and then the structural assignments correlated\nfor characteristic bands as illustrated in the results.<\/p>\n\n\n\n

1H nuclear magnetic resonance spectroscopy (NMR) analysis <\/strong><\/p>\n\n\n\n

The spectra of the NMR were taken by\ndissolving the sample in dimethyl sulfoxide\n(DMSO) \u2013 d6 and later on running the NMR\nSpectrometer. All the chemical shifts were reported with\ntetramethyl silane (TMS) reference at 0 parts\nper million (ppm).  NMR measurement was\ncarried out at Al-Albayt University\/Al-Mafraq\/Jordan and the Euro-vector EA 3000A NMR spectrometer\napparatus (300MHz for 1H-NMR) was used. Chemical shifts are\ngiven on a \u03b4 (ppm) scale with TMS as the internal standard. <\/p>\n\n\n\n

Liquid chromatography\/ mass spectroscopy LC\/MS <\/strong><\/p>\n\n\n\n

Analytical LC\/MS was done by using Shimadzu-LC \/ MS-8040 series system and coupled to a mass Spectrometer-Shimadzu with\nan electrospray interphase (ESI). The caffiec and p-coumaric acids isolated\nfrom Coriandrum sativum <\/em>dried aerial parts were performed by C18, pore\nsize 3.5 \u03bcm, length 15 cm, id 4.6\u03bcm column, the column was maintained at 42C0<\/sup>\ntemperature, the mobile phase was composed of 0.1% acetic acid and acetonitrile\n(V\/V), the flow rate was 0.8 ml\/min and the isocratic mode of elution was used.\nThe volume of the injected standard and sample was 5 \u03bcL and Negative ionization\nmode was applied for structural elucidation 24,25<\/sup>.<\/p>\n\n\n\n

Preparation of the isolated compounds <\/strong><\/p>\n\n\n\n

Two milligrams of caffeic acid and 2\nmg of p-coumaric acid were dissolved in 2 mL of ethanol separately and after\nthat filtrated using disposable filters of 0.45 \u03bcm pore size before utilization\nof LC\/MS for analysis. <\/p>\n\n\n\n

MS conditions<\/strong><\/p>\n\n\n\n

The mass analysis was performed by a\nShimadzu mass spectrometer with an electrospray interphase (ESI) and then Chem Station was used for the\nchromatographic data processing. MS data were acquired in both modes the\nnegative and positive ionization at a defined condition that is: nitrogen gas\nflow rate 3 l\/min, heat block temperature 400 C\u00b0, desolvation line (DL)\ntemperature 250 C\u00b0, drying gas flow rate 15 l\/min and\nmass spectra were recorded using the fill scan mode in the range of 100-800 Daltons\n26<\/sup>.<\/p>\n\n\n\n

Results and Discussion<\/strong><\/p>\n\n\n\n

Caffeic\nand p-coumaric acids are curative phenolic acids that are used in nutrition and medicine. However, recent\nstudies have reported their many pharmacological effects for the treatment of various disorders. Apium graveolens <\/em>(Celery) and\nCoriandrum sativum<\/em>(Coriander)\nare readily available in the supermarket, and they have many phytochemicals that are widely known for their various applications in the medical field\nand other uses.<\/p>\n\n\n\n

The percentage yields (%w\/w) of each ethyl\nacetate fraction for dry and fresh aerial parts of Apium\ngraveolens <\/em>and Coriandrum sativum <\/em>are shown in Table 2.<\/p>\n\n\n\n

Table 2: The percentages of ethyl acetate fractions of dry and fresh aerial parts of Coriandrum sativum<\/em> and Apium graveolens<\/em>.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

Analyzed fraction<\/strong><\/p>\n<\/td>\n

\n

Percentage yield<\/strong><\/p>\n

(%w\/w)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Fresh Coriandrum sativum<\/em> (FC)<\/p>\n<\/td>\n

\n

2.6 %<\/p>\n<\/td>\n<\/tr>\n

\n

Dry Coriandrum sativum<\/em> (DC)<\/p>\n<\/td>\n

\n

4.7 %<\/p>\n<\/td>\n<\/tr>\n

\n

Fresh  Apium graveolens<\/em> (FA)<\/p>\n<\/td>\n

\n

2.4 %<\/p>\n<\/td>\n<\/tr>\n

\n

Dry Apium graveolens<\/em> (DA)<\/p>\n<\/td>\n

\n

3.5 %<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Identification of <\/strong>phenolic acids <\/strong>by TLC<\/strong><\/p>\n\n\n\n

The\nidentification of caffeic and p-coumaric acids\nwas achieved by TLC in aerial\nparts of the ethyl acetate extracts of Apium\ngraveolens <\/em>and Coriandrum <\/em>sativum <\/em>as shown in Figure (2). <\/p>\n\n\n\n

After observation of the spots under UV at 254 nm and by spraying with ferric chloride (0.5%), caffeic and p-coumaric acids in plant samples are detected by calculating their Rf values as shown in Table (3)<\/p>\n\n\n\n

Table 3: The Rf values of separated phenolic acids (caffeic and p-coumaric acids) and their standards in one solvent system in TLC.<\/strong> <\/p>\n\n\n\n\n\n\n\n
\n

Rf value<\/strong><\/p>\n<\/td>\n

\n

Standards<\/strong><\/p>\n<\/td>\n

\n

Fresh Coriandrum sativum<\/em> (FC)<\/strong><\/p>\n<\/td>\n

\n

Dry Coriandrum sativum<\/em> (DC)<\/strong><\/p>\n<\/td>\n

\n

Fresh  Apium graveolens<\/em> (FA)<\/strong><\/p>\n<\/td>\n

\n

Dry Apium graveolens<\/em> (DA)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

caffeic acid<\/strong><\/p>\n<\/td>\n

\n

0.371<\/p>\n<\/td>\n

\n

0.371<\/p>\n<\/td>\n

\n

0.371<\/p>\n<\/td>\n

\n

0.371<\/p>\n<\/td>\n

\n

0.371<\/p>\n<\/td>\n<\/tr>\n

\n

p-coumaric acid<\/strong><\/p>\n<\/td>\n

\n

0.513<\/p>\n<\/td>\n

\n

0.513<\/p>\n<\/td>\n

\n

0.513<\/p>\n<\/td>\n

\n

0.513<\/p>\n<\/td>\n

\n

0.513<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\nFigure 2: TLC of ethyl acetate extracts of fresh and dry aerial parts of Coriandrum sativum<\/em> and Apium graveolens<\/em> using adsorbent (silica gel GF254) and a mobile phase (toluene: ethyl acetate: formic acid (36:12:5)).<\/strong>

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Qualitative\nand quantitative <\/strong>estimations<\/strong> of <\/strong>phenolic acids<\/strong> by HPLC<\/strong><\/p>\n\n\n\n

HPLC\nwas performed to determine the existence and quantity of caffeic\nand p-coumaric acids in the extracts of Apium graveolens <\/em>and Coriandrum <\/em>sativum<\/em> (fresh and dry aerial parts).<\/p>\n\n\n\n

For qualitative estimations, the result revealed that the retention\ntime of p-coumaric and caffeic acids in different ethyl acetate extracts and\nthe retention time of their standards were similar. This proved the presence of\np-coumaric and caffeic acids in the samples as shown in Figures (3-8).<\/p>\n\n\n\n\n\n
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Figure 3: HPLC analysis of caffeic acid standard<\/strong><\/p>\n

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Figure 4: HPLC analysis of p-coumaric acid standard<\/strong><\/p>\n

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Figure 5: HPLC analysis of ethyl acetate fractions of dry Apium graveolens<\/em> <\/strong><\/p>\n

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Figure 6: HPLC analysis of ethyl acetate fractions of fresh Apium graveolens<\/em><\/strong><\/p>\n

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Figure 7: HPLC analysis of ethyl acetate fractions of dry Coriandrum sativum<\/em><\/strong><\/p>\n

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Figure 8: HPLC analysis<\/strong> of <\/strong>ethyl acetate fractions of <\/strong>fresh Coriandrum sativum<\/em> <\/strong><\/p>\n

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Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

In quantitative estimations of p-coumaric and caffeic acids in fractions analyzed from coriander and celery plants by HPLC, a plot of the area vs. standard concentration for both caffeic and p-coumaric acids show a linear fit. The concentration of these phenolic acids in each fraction was determined by using a straight-line equation as in Figures (9,10). <\/p>\n\n\n\n

The coriander and celery plants had shown different concentrations of p-coumaric\nand caffeic acids, as shown in Table 4.<\/p>\n\n\n\n

The results indicated that the\nconcentration of p-coumaric and caffeic acids is higher in the ethyl acetate fraction of the\ndry coriander aerial part (DC) than their concentrations in other ethyl acetate\nfractions. Therefore, p-coumaric and caffeic acids are isolated by preparative\nTLC from the ethyl acetate fraction of DC. <\/p>\n\n\n\n

The result revealed that the HPLC method was efficient for the quantitative and qualitative determination\nof caffeic and p-coumaric acids.<\/p>\n\n\n\n

Table 4: Concentration of caffeic and p-coumaric acids in fresh and dry aerial parts of coriander and celery.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

Ethyl acetate fraction<\/strong><\/p>\n<\/td>\n

\n

The concentration of caffeic acid (ppm)<\/strong><\/p>\n<\/td>\n

\n

Concentration of p-coumaric acid (ppm)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Dry Coriandrum sativum<\/em> (DC)<\/strong><\/p>\n<\/td>\n

\n

166.911<\/p>\n<\/td>\n

\n

247.615<\/p>\n<\/td>\n<\/tr>\n

\n

Fresh Coriandrum sativum<\/em> (FC)<\/strong><\/p>\n<\/td>\n

\n

166.883<\/p>\n<\/td>\n

\n

211.357<\/p>\n<\/td>\n<\/tr>\n

\n

Dry Apium graveolens<\/em> (DA)<\/strong><\/p>\n<\/td>\n

\n

120.878<\/p>\n<\/td>\n

\n

181.256<\/p>\n<\/td>\n<\/tr>\n

\n

Fresh\u00a0 Apium graveolens<\/em> (FA)<\/strong><\/p>\n<\/td>\n

\n

112.365<\/p>\n<\/td>\n

\n

165.021<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 9: HPLC calibration curve of the <\/strong>caffeic acid<\/strong>s standard.<\/strong><\/p>\n

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Figure 10: HPLC calibration curve of the <\/strong>p-coumaric acid<\/strong>s standard.<\/strong><\/p>\n

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Isolation of phenolic acids by preparative TLC plates<\/strong><\/p>\n\n\n\n

The development of the ethyl\nacetate fraction of the aerial part of dry coriander in toluene: ethyl acetate: formic acid\n(36:12:5) in preparative TLC plates results in the number of bands that represent several compounds as in Figure 11.<\/p>\n\n\n\n\n\n
\"\"<\/td>\nFigure 11: Preparative TLC plates of the ethyl acetate fraction of the dry aerial part of coriander (sample = DC) developed in toluene: <\/strong>

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Identification and characterization of the isolated constituents<\/strong><\/p>\n\n\n\n

The isolated constituents that result from preparative TLC were subjected to\ndifferent analysis identification techniques to determine their purity of them\nin addition to other several techniques that were applied\nto obtain the structural elucidation. These techniques include the following:<\/p>\n\n\n\n

FTIR<\/strong><\/p>\n\n\n\n

The IR spectra of isolated caffeic and p-coumaric acids that gave similar results were matched with their standard; which indicated that isolated compounds are p-coumaric and caffeic acids as shown in Figures (12,13).<\/p>\n\n\n\n\n\n
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Figure 12: FTIR spectra of A= isolated caffeic acid and B = standard caffeic acid.<\/strong><\/p>\n

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Figure 13: FTIR spectra of A= isolated p-coumaric acid and B= p-coumaric acid standard.<\/strong><\/p>\n

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The isolated compounds’ characteristic\nFTIR absorption bands with their standards (p-coumaric and caffeic acids) are\nlisted in Tables 5 and 6, respectively27,28<\/sup>.<\/p>\n\n\n\n\n\n
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Table 5: FTIR absorption bands (cm-1<\/sup>) of the isolated caffeic acid.<\/strong><\/p>\n

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Table 6: FTIR (cm-1) absorption bands of isolated p-coumaric acid.<\/strong><\/p>\n

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Nuclear magnetic resonance (NMR) spectroscopy<\/strong><\/p>\n\n\n\n

Is a sophisticated and powerful\nanalytical technique for the elucidation of organic compound structures and\nidentification of molecular interaction 28<\/sup>. <\/p>\n\n\n\n

For that reason,\nthe identification of isolated, p-coumaric, and caffeic acids was further confirmed by NMR spectroscopy\napplication. The isolated constituents’ NMR spectra are in good agreement with works of\nliterature on p-coumaric and caffeic acids as shown in Figures 14,15 and\nTables 7 and 8 26,29,30<\/sup>.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Table 7: Assignment of <\/strong>1<\/strong>H-NMR spectral data of isolated caffeic acid.<\/strong><\/p>\n

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Click here to view Table<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

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Figure 14: <\/strong>1<\/strong>H-NMR spectrum of isolated caffeic acid.<\/strong><\/p>\n

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Table 8: Assignment of <\/strong>1<\/strong>H-NMR spectral data of isolated p-coumaric acid.<\/strong><\/p>\n

\u00a0<\/p>\n

Click here to view Table<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

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Figure 15: 1H-NMR spectrum of isolated p-coumaric acid.<\/strong><\/p>\n

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LC\/MS<\/strong><\/p>\n\n\n\n

The identification of isolated caffeic acid and p-coumaric acids were further confirmed by LC\/MS. For structural scanning, full scan product ion was used instead of multiple reaction monitoring (MRM) since it was used for qualitative applications to obtain structural information. <\/p>\n\n\n\n

According to the chromatographic\nconditions, Rt (retention times) of the analyzed constituents are 5.6 min for caffeic acid and 7.8 min for p-coumaric acid; in addition, the ions monitored by the MS method are illustrated in Table 9. <\/p>\n\n\n\n

In the ionization conditions, most of the phenolic acids lost a\nmolecule of COO (44 units) so ions detected by spectrometer are always in the\nform of [M-H-COO] \u0305. All these data are in good agreement\nwith the works of literature on p-coumaric and\ncaffeic acids31-33<\/sup>. <\/p>\n\n\n\n

Table 9: The specific ion of isolated caffeic acid and p-coumaric acid was monitored in the screening method.<\/strong><\/p>\n\n\n\n\n\n\n\n
\n

Isolated compounds<\/strong><\/p>\n<\/td>\n

\n

Rt<\/sub> (min)<\/strong><\/p>\n<\/td>\n

\n

theoretical molecular weight (M)<\/strong><\/p>\n<\/td>\n

\n

Calculated molecular weight (M)<\/strong><\/p>\n<\/td>\n

\n

Fragmented ions (m\/z)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

caffeic acid<\/p>\n<\/td>\n

\n

5.6<\/p>\n<\/td>\n

\n

180.16<\/p>\n<\/td>\n

\n

180<\/p>\n<\/td>\n

\n

179(M-H), 135(M-H-CO2<\/sub>), 161(M-H-H2<\/sub>O), 134(M-H-HCO3<\/sub>)<\/p>\n<\/td>\n<\/tr>\n

\n

p-coumaric acid<\/p>\n<\/td>\n

\n

8.7<\/p>\n<\/td>\n

\n

164.158<\/p>\n<\/td>\n

\n

164<\/p>\n<\/td>\n

\n

163(M-H), 119(M-H-CO2<\/sub>),113, and 101 (dissociation of the phenolic ring)<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

These assignments were proved by\nfull product ion scan liquid chromatography connected with\na negative (-ve) ES ionization spectrum of caffeic acid (Figure 16) and p-coumaric\nacid (Figure 17).<\/p>\n\n\n\n\n\n
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Figure 16: Full product ion scan LC chromatogram and mass spectrum of isolated caffeic acid.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 17: Full product ion scan LC chromatogram and mass spectrum of isolated p-coumaric acid. <\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

Coriander\n(Coriandrum sativum<\/em> L.) and celery (Apium graveolens<\/em> L.) are\nexcellent sources of caffeic and p-coumaric acids (phenolic acids).\nThese compounds are considered a substantial class of natural components that are widely used in several countries as an ingredient in\nmany herbal remedies and preparation. \nThis work shows that the aerial parts extracts of the coriander and celery plant contain caffeic and p-coumaric acids, and quantitative estimation by HPLC shows that the concentrations of caffeic and p-coumaric acids in the dry coriander are higher than in the fresh coriander and celery (dry and\nfresh). According to the HPLC result, the caffeic and p-coumaric acids were\nisolated from dry coriander by using the preparative TLC method. This study\nconfirms that coriander showed significantly higher p-coumaric\nand caffeic acids contents than celery.<\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

None<\/p>\n\n\n\n

Conflict on Interest<\/strong><\/p>\n\n\n\n

There is no conflict of interest<\/p>\n\n\n\n

Funding Source<\/strong><\/p>\n\n\n\n

There are no funding sources.-<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

  1. Nhut PT, Quyen NT, Truc TT, Minh LV, An TN, Anh NH. Preliminary study on phytochemical, phenolic content, flavonoids, and antioxidant activity of Coriandrum sativum<\/em> L. originating in Vietnam. InIOP Conference Series: Materials Science and Engineering<\/em> 2020;991(1):012-022.<\/li>
  2. Bhat S, Kaushal P, Kaur M, Sharma K. Coriander (Coriandrum sativum <\/em>L.): Processing, nutritional and functional aspects. Afr J Plant Sci<\/em> 2014; 8:5-33.<\/li>
  3. Pathak NL, Kasture SB, Bhatt NM. Phytochemical screening of Coriander sativum <\/em>Linn. Int J Pharm Sci Rev Res<\/em> 2011; 9:159-163.<\/li>
  4. Ahmed EH, Abadi RS, Mohammed AM. Phytochemical screening, chemical composition and antioxidant activity of seeds essential oil of Coriandrum sativum <\/em>L. from the Sudan.<\/em> International Journal of Herbal Medicine<\/em> 2018; 6(1): 01-04.<\/li>
  5. Karmakar UK, Rahman MA, Roy DN, Sadhu SK, Ali ME. Chemical and biological investigations of Coriandrum sativum<\/em> Linn. IJPSR<\/em> 2011; 2(4): 999-1006.<\/li>
  6. Ghani A.  Medicinal plants of Bangladesh, 2nd Ed. The Asiatic Society of Bangladesh, Dhaka, 2003; pp 382.<\/li>
  7. Rathore SS, Saxena SN, Balraj S. Potential health benefits of major seed spices. Int J Seed Spices<\/em> 2013; 3:1-12.<\/li>
  8. Cortes-Eslava J, Gomez-Arroyo S, Villalobos-Pietrini R. Antimutagenicity of Coriander (Coriandrum sativum<\/em>) juice on the mutagenesis produced by plant metabolites of aromatic amines. J Toxicol Lett<\/em> 2004; 153:283-292.<\/li>
  9. Eidi M, Eidi A, Saeidi A, Molanaei S, Sadeghipour A, Bahar M et al. <\/em>Effect of Coriander seed (Coriandrum sativum <\/em>L) ethanol extract on insulin release from pancreatic beta cells in streptozotocin-induced diabetic rats. J Phytother Res<\/em> 2012; 23:404-406.<\/li>
  10.  Alison MG, Peter RF. Insulin-releasing and insulin-like activity of the traditional antidiabetic plant Coriander sativum <\/em>(coriander). British J Nutr<\/em>1999; 81:203-209.<\/li>
  11.  Sunil C, Agastian P, Kumarappan C, Ignacimuthu S. In vitro <\/em>antioxidant, antidiabetic and antilipidemic activities of Symplocos cochinchinensis <\/em>(Lour.) S. Moore bark. J Food Chem Toxicol<\/em> 2012; 50:1547-1553. <\/li>
  12.  Nair SN, Varghese A, Meenu B, Rejimon G, Neeraja E D. Comparative evaluation of Coriandrum sativum Linn. <\/em>and Apium graveolens <\/em>for Antimicrobial activity. Research J. Pharm and Tech <\/em>2017; 10(2):541-544.<\/li>
  13.  Sarmanovna TZ. Phytochemical Study of Odorous Celery Root (Apium graveolens <\/em>L.) Grown in the North Caucasus. Pharmacogn J<\/em> 2019; 11(3): 527-530.<\/li>
  14.  Uddin Z, Shad AA, Bakht J, Ullah I, Jan S. In vitro antimicrobial, antioxidant activity, and phytochemical screening of Apium graveolens<\/em>. Pak J Pharm Sci<\/em> 2015; 28 (5):1699-1704.<\/li>
  15.  Popovic M, Kaurinovic B, Triviv S, Mimica-Dukic N, Bursac M.  Effect of celery (Apium graeolens<\/em>) extracts on some biochemical parameters of oxidative stress in mice treated with carbon tetrachloride. Phytother<\/em> 2006; 20: 531-536.<\/li>
  16.  Edziri H, Ammar S, Souad L, Mahjoub MA, Aouni M, Mighri Z, Verschaeve L. In vitro <\/em>evaluation of antimicrobial and antioxidant activities of some Tunisian vegetables. South Afri J Bot<\/em> 2012; 78: 252-256.<\/li>
  17.  Shad AA, Shah HU, Bakht J, Choudhary MI, Ullah J. Nutraceutical potential and bioassay of Apium graveolens <\/em>L. grown in Khyber Pakhtunkhwa-Pakistan. J Med Plant Res<\/em> 2011; 5: 5160-5166.<\/li>
  18.  Asgarpanah J and Kazemivash N. Phytochemistry, pharmacology and medicinal properties of Coriandrum sativum <\/em>L. African Journal of Pharmacy and Pharmacology<\/em> 2012; 6(31): 2340-2345.<\/li>
  19.  R.A. Mans D, Grant A, Pinas N. Plant-Based Ethnopharmacological Remedies for Hypertension in Suriname. Herbal Medicine<\/em> 2017.<\/li>
  20.  <\/strong>Iswantini D, Ramdhani TH, Darusman LK. In vitro, <\/em>inhibition of celery (Apium graveolens <\/em>l.) extract on the activity of xanthine oxidase and determination of its active compound. Indo J Chem <\/em>2012, 12 (3): 247-254.<\/li>
  21.  Medic-Saric M, Jasprica I, Smoicic-Bubalo A, Mornar A. Optimization of chromatographic conditions in thin layer. Croatica Chemica Acta<\/em> 2004; 77(1-2):361-366.<\/li>
  22. Shah Sh M, Abdul-Jalil Th Z. Qualitative and Quantitative Estimation or Chemical Constituents from Leaves and Roots of Iraqi Agave attenuata<\/em> by GC-MS and RP-HPLC. Iraqi J Pharm Sci<\/em> 2022; 31(Sppl):75-85.<\/li>
  23.  Dubber MJ, Kanfer I. High-performance liquid chromatographic determination of selected flavonols in Ginkgo biloba<\/em> solid oral dosage forms. J Pharm Pharm<\/em> Sci<\/em> 2004;7(3):303-309.<\/li>
  24.  Adams M, Weidenmann M, Tittel G, Bauer R. HPLC-MS trace analysis of atropine in Lycium barbarum <\/em>berries. Phyto Chem Anal<\/em> 2006; 17: 279-283.<\/li>
  25.  MoCan A, Vlase L, Letitia AA, Vodnar D, Bischin C, Dumitrescu R.S., Crisan G. HPLC\/MS analysis of Caffeic and chlorogenic acids from three Romanian Veronica species and their antioxidant and antimicrobial properties. Farmacia <\/em>2015; 63(6): 890-896. <\/li>
  26.  Kajdzanoska M, Gjamovski V, Stefova M.  HPLC-DAD-ESI-MS Identification of phenolic compounds in cultivated Strawberry from Macedonia. Maced J Chem<\/em> Chem Eng<\/em> 2010; 29(2): 181-194. <\/li>
  27. Al-Hussaniy HA, Al-Kuraishy HM, Abdulameer AG. The Use of Panax Ginseng to Reduce the Cardiotoxicity of Doxorubicin and Study its Effect on Modulating Oxidative Stress, Inflammatory, and Apoptosis Pathways. Macedonian Journal of<\/em> Medical Sciences<\/em> 2022; 10:715-719.<\/li>
  28. Viswannth A, Rao L and Babu P. Simple isolation and characterization of p-coumaric acid from Cynodon dectylon <\/em>Linn (pers). IJPAR<\/em> 2017; 6(2): 314-318.<\/li>
  29.  Wong KC. Review of NMR spectroscopy: basic principle, concepts and application in chemistry. J Chem Educ<\/em> 2014; 91: 1103-1104. <\/li>
  30.  Lopez-Martinez LM, Ortega HS, Navarro RE, Mundo RR, Aguilar GC. A 1<\/sup>HNMR investigation of the interaction between phenolic acids found in Mango (Manguifera indica <\/em>VC Ataulfo) and Papaya (Carica papaya <\/em>CV Maradol) and 1,1-diphenyl-2,2 picrylhydrazal (DPPH) free radicals. PLOS One<\/em> 2015; 11: 1-11.<\/li>
  31.  Swislocka R, Sadowy MK, Kalinowska M, Lewandowski W. Spectroscopic (FTIR, FT-Raman, 1H and 13C NMR and theoretical studies) of p-coumaric acid and alkali metal p-coumarates. Spectroscopy<\/em> 2012; 27: 35-48.<\/li>
  32.  Hossain MB, Rai DK, Brunton NP, Martin-Diana AB, Barry-Ryan C. Characterization of phenolic composition in Lamiaceae species by LC-ESI-MS\/MS. J Agric Food Chem<\/em> 2010; 58(19): 10576-10581.<\/li>
  33.  Wv Z, Ma X, Fang D, Qi H, Ren W, Zhang G. Analysis of caffeic acid derivative from Osmanthus xunnanensis <\/em>using electrospray ionization quadrupole time-of-flight mass spectrometry. EJMS<\/em> 2009; 15:415-429.<\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

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