MOJ Food Processing & Technology

Isolation and Identification of New Alkaloids from

Purslane (Portulacaoleracea L.)Leaves Using HPLC/ESI-
MS

Abstract Research Article

Purslane (Portulacaoleracea L.) leaves are well-known traditional Kurdish favorite Volume 2 Issue 4 - 2016
foods. In the present study, from Purslane leaves ten alkaloids were extracted,
isolated and Identified for the first time, By using HPLC/ESI-MS. The identified
alkaloids were; Nandigerine(1), Reticuline(2), (R)-3,4-Dehydromagnocurarine(3),
itingensine(4), (S)-Tembetarine(5), Homolycorine(6), Angustureine (7), 1
University of Raparin, Chemistry Department, Iraq
Cyclobullatine-A (8), 10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2- 2
Koya University, Chemistry Department, Iraq
one(9) and 10-(5-p-coumaroyl-6-methylpiperidin-2-yl)tetradecan-2-one(10).

Keywords: Purslane; Leaves of Purslane; Alkaloids; HPLC/ESI-MS *Corresponding author: Dara Muhammad Aziz,
University of Raparin, Chemistry Department, Iraq, Tel:
+9647510571588; Email:

Received: May 22, 2016 | Published: June 30, 2016

Introduction Materials and Methods

Alkaloids are naturally occurring chemical compounds Plant material
containing basic nitrogen atoms. Alkaloids are produced by a large
variety of organisms, including bacteria, fungi, plants and animals. Purslane Leaves were collected in Rania - Kurdistan Region in
Many alkaloids often have pharmacological effects and are used northern of Iraq, in April 2015. The specimen taxonomically was
as medications, as recreational drugs or in Entheogenic rituals. identified in the Biology Department at University of Raparin.
Over 10000 alkaloids have been isolated from nature so far, which Fresh leaves were dried in open air without exposure to direct
are distributed widely in plants [1]. Many studies demonstrate sunlight. The dried sample was powdered and weighed.
that alkaloids had many kinds of biological activities, such as anti- Chemicals and reagents
microbial, anti-oxidants, anti-cancer, anti-inflammatory, and anti-
virus activities [2,4]. All chemicals and solvents used were in HPLC grade form
Sigma-Aldrich (UK).
The Purslane (Portulacaoleracea L.) is listed in the World
Health Organization (WHO) as one of the most used medicinal Extraction and isolation of alkaloids: Dried powdered Purslane
plants under the family Portulacaceae [5-6]. The genus Portulacao leaves (1000 g) immersed in glacial acetic acid (1000 ml) at room
comprising about 70 species is characterized by conspicuously temperature for overnight. The extract was filtered to remove any
fleshy sessile leaves [7]. residue from the solution. Then, the extract was diluted with de-
ionized water. The extract fractionated with 1000 ml chloroform
Many varieties of Purslane under many names grow in a wide in a separatory funnel (21000 ml). The chloroform extracts were
range of climates and regions, it can be found in Europe, Africa, combined and washed with 10% aqueous sodium bicarbonate.
North America, Australia and Asia. The chloroform extract was dried over anhydrous Sodium Sulfate
Purslane has been ranked as eight Most common plants in the then the solvents were evaporated on a rotary evaporator with
world and is widespread as a weed, fast growing, self-compatible water bath, temperature set up at 45 C.
and has amazing ability to produce seeds even on deaths doorstep Instrumentation HPLC/ESI-MS: The chloroform extract (100
[8,9]. There are many types of Purslane available in Kurdistan mg) dissolved in 10 ml of methanol and filtered through micro-
and mostly still morphologically under investigation. Different filter (40l), then ejected into HPLC/ESI-MS. The samples were
varieties, harvesting times and environmental conditions can analyzed using HPLC/ESI-MS (Aligent 1100 series). The system
contribute to purslanes nutritional composition and benefits was controlled with Chemstation for LC 3D Rev A.09.03. The
[10]. chromatographic separations were performed on the column
The present study focused on isolation and identification of (Spherisil ODS 2 C18, 25 x 4.6 m, 5 um, Waters) to separate
alkaloids from Purslane leaves [1]. Solvent extraction will be used alkaloids. The column temperature was 25 C. The solvent system
to extract the alkaloids [11], and then the extract will be ejected was used a gradient of methanol (A) and water (B), running
into HPLC/ESI-MS. gradient was 90:10 %. The flow rate was 1 ml/min of solvents for
20 minutes and the injection volume of samples and standards was

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Isolation and Identification of New Alkaloids from Purslane (Portulacaoleracea L.)Leaves Copyright:
2016 Aziz et al. 2/4
Using HPLC/ESI-MS

20 l. The detection of the system was combined with diode array Results and Discussion
detector (DAD). The chromatography analysis was developed to
obtain a short retention time of analysis with good resolution. The Purslane leaves extract was dissolved in methanol then
All the chromatograms were recorded at 254 nm for aromatic ejected into HPLC/ESI-MS. Overall 10 alkaloids were analyzed,
system, 280 nm for cinnamic acids and 330 nm for poly phenolics. the HPLC chromatogram shown in Figure 1. The HPLC fitted
For identification the individual alkaloids, the MS analysis was with ESI-MS system to provide molecular mass and further
carried out with an ESI interface operating the positive ion mode structural information of all peaks in the HPLC chromatogram. In
[M+H+]. The MS operating conditions were as follows: Ion spray the Purslane leaves, the presence of the following alkaloids were
voltage, 1.6eV; curtain gas (N2), 20 Psi; nebulizing gas and heating confirmed; Nandigerine (peak no.1), Reticuline (2) Nandigerine
gas (N2), 50 Psi; heating gas temperature, 550 C; SQ detector; (3) Gitingensine (4) (S)-Tembetarine (5) Homolycorine (6)
spectra range, m/z 200 - 900 (scan time, 4.8 sec).The system was Angustureine (7) Cyclobullatine-A (8)10-(5-p-coumaroyl-6-
controlled with Xcalibur software, version 1.2. methylpiperidin-2-yl) dodecan-2-one(9) 10-(5-p-coumaroyl-6-
methylpiperidin-2-yl)tetradecan-2-one (10) (Table 1).

Table 1: Retention time and ESI/MS fragmentation of Alkaloids identified in Purslane leaves.

Retention Time
Peak Number Proposed compounds [M+H]+ (m/z) Fragment Ion (m/z)
(Minutes)

1 0.05 Nandigerine 312 295,280

2 0.117 Reticuline 330 299,213

3 3.377 (R)-3,4-Dehydromagnocurarine 312 297,285,

4 4.553 Gitingensine 329 236,221

5 7.066 (S)-Tembetarine 344 299,250

6 8.531 Homolycorine 316 302,298

7 10.764 Angustureine 217 217,202

8 12.11 Cyclobullatine-A 414 338,324

10-(5-p-coumaroyl-6-methylpiperidin-
9 14.184 441 298,280
2-yl) dodecan-2-one

10-(5-p-coumaroyl-6-methylpiperidin-
10 19.187 472 326,308
2-yl) tetradecan-2-one

From the MS spectrum and by comparing with previous time of reporting those alkaloids in Purslane leaves. Alkaloids in
reports the identification of the alkaloids were confirmed [12,13]. peaks 5,6,7,8,9 and 10 assigned as (S)-Tembetarine, Homolycorine,
In the mass spectrum (Figure 2), showed a protonated ion at m/z Angustureine, Cyclobullatine-A, 10-(5-p-coumaroyl-6-
312, which is match to Nandigerine (1) in the chromatogram, (the methylpiperidin-2-yl) dodecan-2-one and 10-(5-p-coumaroyl-
chemical structure are corresponded with numbers in Figure 6-methylpiperidin-2-yl) tetradecan-2-one respectively. The
3. In the MS/MS spectrum, [M+H+] ion of Nandigerine observed alkaloids structures were tentatively identified based on their MS
two fragment ions at 280 and 295 m/z, protonated Nandigerine fragmentation behavior, as shown in Table 1.
previously was reported with the same condition [14]. Also, the
fragmentation behaviors of protonated Reticuline (2), (R)-3,4- Conclusion
Dehydromagnocurarine (3) and Gitingensine (4) were found and Leaves of Purslane (Portulacaoleracea L.) were extracted with
published with the same condition [15-17]. glacial Acetic Acid and fractionated with Chloroform to extract
The last 6 alkaloids were reported or tentatively characterized alkaloids. Then, the extract ejected into HPLC/ESI-MS to analysis
in other species [18-23]. But as far as authors concerned this first and confirm the chemical structure of alkaloids. In Purslane leaves,

Citation: Aziz DM, Taher SG, Hama JR (2016) Isolation and Identification of New Alkaloids from Purslane (Portulacaoleracea L.)Leaves Using HPLC/
ESI-MS. MOJ Food process Technol 2(4): 00047. DOI: 10.15406/mojfpt.2016.02.00047
Isolation and Identification of New Alkaloids from Purslane (Portulacaoleracea L.)Leaves Copyright:
2016 Aziz et al. 3/4
Using HPLC/ESI-MS

the following alkaloids were identified; which are; Nandigerine, Acknowledgment

Reticuline, (R)-3,4-Dehydromagnocurarine, Gitingensine, (S)-
Tembetarine, Homolycorine, Angustureine, Cyclobullatine-A, The authors would like to thank technical staff of Chemistry
10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2-one and Department and Faculty of Science at University of Raparin, also
10-(5-p-coumaroyl-6-methylpiperidin-2-yl)tetradecan-2-one. to Koya University to let us do the practical work in their chemical
laboratories. This work was financed with grants from HCDP
program in Ministry of Higher Education and Scientific Research
of Kurdistan - Iraq, 2015, no. JUR16.

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ESI-MS. MOJ Food process Technol 2(4): 00047. DOI: 10.15406/mojfpt.2016.02.00047
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Using HPLC/ESI-MS

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Citation: Aziz DM, Taher SG, Hama JR (2016) Isolation and Identification of New Alkaloids from Purslane (Portulacaoleracea L.)Leaves Using HPLC/
ESI-MS. MOJ Food process Technol 2(4): 00047. DOI: 10.15406/mojfpt.2016.02.00047