Food Processing & Technology
Research Article
Volume 2 Issue 4 - 2016
Isolation and Identification of New Alkaloids from Purslane (Portulacaoleracea L.)Leaves Using HPLC/ESI-MS
Dara Muhammed Aziz1* Salam Ghafour Taher2 and Jawameer Rasool Hama1
1University of Raparin, Chemistry Department, Iraq
2Koya University, Chemistry Department, Iraq
Received: May 22, 2016 | Published: June 30, 2016
*Corresponding author: Dara Muhammad Aziz, University of Raparin, Chemistry Department, Iraq, Tel: +9647510571588; Email:
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


Purslane (Portulacaoleracea L.) leaves are well-known traditional Kurdish favorite 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), Cyclobullatine-A (8), 10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2-one(9) and 10-(5-p-coumaroyl-6-methylpiperidin-2-yl)tetradecan-2-one(10).

Keywords: Purslane; Leaves of Purslane; Alkaloids; HPLC/ESI-MS


Alkaloids are naturally occurring chemical compounds containing basic nitrogen atoms. Alkaloids are produced by a large variety of organisms, including bacteria, fungi, plants and animals. Many alkaloids often have pharmacological effects and are used as medications, as recreational drugs or in Entheogenic rituals. Over 10000 alkaloids have been isolated from nature so far, which are distributed widely in plants [1]. Many studies demonstrate that alkaloids had many kinds of biological activities, such as anti-microbial, anti-oxidants, anti-cancer, anti-inflammatory, and anti-virus activities [2,4].

The Purslane (Portulacaoleracea L.) is listed in the World Health Organization (WHO) as one of the most used medicinal plants under the family Portulacaceae [5-6]. The genus Portulacao comprising about 70 species is characterized by conspicuously fleshy sessile leaves [7].

Many varieties of Purslane under many names grow in a wide range of climates and regions, it can be found in Europe, Africa, North America, Australia and Asia.

Purslane has been ranked as eight Most common plants in the world and is widespread as a weed, fast growing, self-compatible and has amazing ability to produce seeds even on death’s doorstep [8,9]. There are many types of Purslane available in Kurdistan and mostly still morphologically under investigation. Different varieties, harvesting times and environmental conditions can contribute to purslane’s nutritional composition and benefits [10].

The present study focused on isolation and identification of alkaloids from Purslane leaves [1]. Solvent extraction will be used to extract the alkaloids [11], and then the extract will be ejected into HPLC/ESI-MS.

Materials and Methods

Plant material

Purslane Leaves were collected in Rania - Kurdistan Region in northern of Iraq, in April 2015. The specimen taxonomically was identified in the Biology Department at University of Raparin. Fresh leaves were dried in open air without exposure to direct sunlight. The dried sample was powdered and weighed.

Chemicals and reagents

All chemicals and solvents used were in HPLC grade form Sigma-Aldrich (UK).

Extraction and isolation of alkaloids: Dried powdered Purslane leaves (1000 g) immersed in glacial acetic acid (1000 ml) at room temperature for overnight. The extract was filtered to remove any residue from the solution. Then, the extract was diluted with de-ionized water. The extract fractionated with 1000 ml chloroform in a separatory funnel (2×1000 ml). The chloroform extracts were combined and washed with 10% aqueous sodium bicarbonate. The chloroform extract was dried over anhydrous Sodium Sulfate then the solvents were evaporated on a rotary evaporator with water bath, temperature set up at 45 °C.

Instrumentation HPLC/ESI-MS: The chloroform extract (100 mg) dissolved in 10 ml of methanol and filtered through micro-filter (40μl), then ejected into HPLC/ESI-MS. The samples were analyzed using HPLC/ESI-MS (Aligent 1100 series). The system was controlled with Chemstation for LC 3D Rev A.09.03. The chromatographic separations were performed on the column (Spherisil ODS 2 C18, 25 x 4.6 m, 5 um, Waters) to separate alkaloids. The column temperature was 25 °C. The solvent system was used a gradient of methanol (A) and water (B), running 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 20 μl. The detection of the system was combined with diode array detector (DAD). The chromatography analysis was developed to obtain a short retention time of analysis with good resolution. All the chromatograms were recorded at 254 nm for aromatic system, 280 nm for cinnamic acids and 330 nm for poly phenolics. For identification the individual alkaloids, the MS analysis was carried out with an ESI interface operating the positive ion mode [M+H+]. The MS operating conditions were as follows: Ion spray voltage, 1.6eV; curtain gas (N2), 20 Psi; nebulizing gas and heating gas (N2), 50 Psi; heating gas temperature, 550 ËšC; SQ detector; spectra range, m/z 200 - 900 (scan time, 4.8 sec).The system was controlled with Xcalibur software, version 1.2.

Results and Discussion

The Purslane leaves extract was dissolved in methanol then ejected into HPLC/ESI-MS. Overall 10 alkaloids were analyzed, the HPLC chromatogram shown in Figure 1. The HPLC fitted with ESI-MS system to provide molecular mass and further structural information of all peaks in the HPLC chromatogram. In the Purslane leaves, the presence of the following alkaloids were confirmed; Nandigerine (peak no.1), Reticuline (2) Nandigerine (3) Gitingensine (4) (S)-Tembetarine (5) Homolycorine (6) Angustureine (7) Cyclobullatine-A (8)10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2-one(9) 10-(5-p-coumaroyl-6-methylpiperidin-2-yl)tetradecan-2-one (10) (Table 1).

Peak Number

Retention Time (Minutes)

Proposed compounds

[M+H]+ (m/z)

Fragment Ion (m/z)











































10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2-one





10-(5-p-coumaroyl-6-methylpiperidin-2-yl) tetradecan-2-one



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

Figure 1: HPLC Chromatogram of alkaloids extract from Purslane leaves, each peak refer to a number of an alkaloids listed in Table 1: Nandigerine (1), Reticuline (2), Nandigerine (3), Gitingensine (4), (S)-Tembetarine (5), Homolycorine (6), Angustureine (7), Cyclobullatine-A (8), 10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2-one (9) and 10-(5-p-coumaroyl-6-methylpiperidin-2-yl)tetradecan-2-one (10).

From the MS spectrum and by comparing with previous reports the identification of the alkaloids were confirmed [12,13]. In the mass spectrum (Figure 2), showed a protonated ion at m/z 312, which is match to Nandigerine (1) in the chromatogram, (the chemical structure are corresponded with numbers in Figure 3. In the MS/MS spectrum, [M+H+] ion of Nandigerine observed two fragment ions at 280 and 295 m/z, protonated Nandigerine previously was reported with the same condition [14]. Also, the fragmentation behaviors of protonated Reticuline (2), (R)-3,4-Dehydromagnocurarine (3) and Gitingensine (4) were found and published with the same condition [15-17].

Figure 2: HPLC/ESI-MS spectrum of alkaloid extracts from Purslane leaves.
Figure 3: Chemical structures of alkaloids from Purslane leaves, listed in Table 1.

The last 6 alkaloids were reported or tentatively characterized in other species [18-23]. But as far as authors concerned this first time of reporting those alkaloids in Purslane leaves. Alkaloids in peaks 5,6,7,8,9 and 10 assigned as (S)-Tembetarine, Homolycorine, Angustureine, Cyclobullatine-A, 10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2-one and 10-(5-p-coumaroyl-6-methylpiperidin-2-yl) tetradecan-2-one respectively. The alkaloids structures were tentatively identified based on their MS fragmentation behavior, as shown in Table 1.


Leaves of Purslane (Portulacaoleracea L.) were extracted with glacial Acetic Acid and fractionated with Chloroform to extract alkaloids. Then, the extract ejected into HPLC/ESI-MS to analysis and confirm the chemical structure of alkaloids. In Purslane leaves, the following alkaloids were identified; which are; Nandigerine, Reticuline, (R)-3,4-Dehydromagnocurarine, Gitingensine, (S)-Tembetarine, Homolycorine, Angustureine, Cyclobullatine-A, 10-(5-p-coumaroyl-6-methylpiperidin-2-yl) dodecan-2-one and 10-(5-p-coumaroyl-6-methylpiperidin-2-yl)tetradecan-2-one.


The authors would like to thank technical staff of Chemistry Department and Faculty of Science at University of Raparin, also 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.


  1. Aziz DM, Wso MA, Ibrahim BM (2015) Antimicrobial and antioxidant activities of extracts from medicinal plant ginger (Zingiber officinale) and identification of components by gas chromatography. African Journal of Plant Science 9(10): 412-420.
  2. Yan D, Jin C, Xiao XH, Dong XP (2008) Antimicrobial properties of berberines alkaloids in Coptischinensis Franch by microcalorimetry. J Biochem Biophys Methods 70(6): 845-849.
  3. Liang M,J Zhang, WD, Hua J, Liu RH, Zhang C (2006) Simultaneous analysis of alkaloids from Zanthoxylumnitidumby high performance liquid chromatography-diode array detector-electrospray tandem mass spectrometry. J Pharm Biomed Anal 42(2): 178-183.
  4. Zhou H, Tai YP, Sun CR, Pan Y (2005) Rapid identification of vinca alkaloids by direct-injection electrospray ionisation tandem mass spectrometry and confirmation by high-performance liquid chromatography-mass spectrometry. Phytochem Anal 16(5): 328-333.
  5. Dweck AC (2001) Purslane (Portulacaoleracea) - The global panacea. Personal Care Magazine 2(4): 7-15.
  6. Ala MA, Juraimi AS, Rafii MY, Hamid A, A Aslani, et al. (2014) Evaluation of antioxidant compounds antioxidant activities and mineral composition of 13 collected purslane (Portulacaoleracea L.) accessions. Bio Med Research International 1: 1-10.
  7. Oliveira I, Valentão P, Lopes R, Andrade P, B Bento, et al. (2009) Phytochemical characterization and radical scavenging activity of Portulacaoleracea L.leaves and stems. Microchemical Journal 92: 129-134.
  8. Liu L, Peter H, Ye-Fang, Z Zhi-Qiang, X Charles, et al. (2000) Fatty acids and b-carotene in Australian Purslane (Portulacaoleracea) varieties. J Chromatogr A 893: 207-213.
  9. Rashed AN, Afifi FU, Disi AM (2003) Simple evaluation of the wound healing activity of a crude extract of Portulacaoleracea L. (growing in Jordan) in Musmusculus JVI- 1. J Ethnopharmacol 88(2-3): 131-136.
  10. Uddin MK, Juraimi AS, Ali ME, Ismail MR (2012) Evaluation of antioxidant properties and mineral composition of Portulacaoleracea (L.) at different growth stages. Int J Mol Sci 13(8): 10257-10267.
  11. Dara MA, Jwameer RH, Alam SM (2015) Synthesizing a novel derivatives of piperine from black pepper (Piper nigrum L.). Journal of Food Measurement and Characterization 9(3): 324-331.
  12. Kihara M, Konishi K, Xu L Kobayashi S (1991) Alkaloidal constituents of the flowers of Lycorisradiata HERB (Amaryllidaceae).Chemical & Pharmaceutical Bulletin 39: 1849-1853.
  13. Wu X, Prior RL (2005) Systematic identification and characterization of anthocyanins by HPLC-ESI-MS/ MS in common foods in the United States Fruits and ber-ries. J Agric Food Chem 53: 2589- 2599.
  14. Moriyasu M, Wang J, Zhang H, Lu GB Ichimaru, et al. (1996) Isolation of alkaloids from plant materials by combination of ion-pair extraction and preparative ion-pair HPLC using sodium perchlorate. Chinese Magnoliae cortex Nat Med 50: 413-416.
  15. Yu SX, Yan RY, Liang RX, Wang W, Yang B (2012) Bioactive polar compounds from stem bark of Magnolia officinalis. Fitoterapia 83: 356-361.
  16. Lu Z,M Zhang Q,J Chen R,Y Yu DQ (2011) Study on chemical constituents from branches and leaves of Polyalthianemoralis. China Journal of Chinese Materia Medica 36: 1024-1027.
  17. Gharneh HA , Hassandokht MR (2012) Chemical composition of some Iranian purslane (Portulacaoleracea) as a leafy vegetable in south parts of Iran. ActaHorticulturae 944: 41-44.
  18. Samy RP, Ignacimuthu S (2000) Antibacterial activity of some folklore medicinal plants used by tribals in Western Ghats of India. J Ethnopharmacol 69(1): 63-71.
  19. Sriphong L, Sotanaphun U, Limsirichalkul S, Wetwitayaklung P, Chaichantipyuth C, et al. (2003) Cytotoxic Alkaloids from the Flowers of Senna spectabilis. Planta Med 69(11): 1054-1056.
  20. SansoresP P, Rosado VM, Brito LW, Mena RGJ Quijano L (2000) Cassine an antimicrobial alkaloid from Sennaracemosa Fitoterapia 71(6): 690-692.
  21. Yoo DS, Choi YH, Cha MR, Lee B H, Kim SJ , et al. (2011) HPLC-ELSD analysis of 18 platycosides from balloon flower roots (Platycodi Ra- dix) sourced from various regions in Korea and geogra- phical clustering of the cultivation areas. Food Chemistry 129: 645-651.
  22. Wang WB, Lu SM, Yang PY, Han XW, Zhou YG (2003) Highly enantioselective iridium-Catalyzed hydrogenation of heteroaromatic Compounds, Quinolines. J Am Chem Soc 125(35): 10536-10537.
  23. Le thi ThH, Votický Z, Paulík V (1981) Alkaloids of Buxussempervirens var rotundifolia BAILLON. Collection of Czechoslovak Chemical Communications 46: 1425-1432.
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