PHYTOCHEMISTRY STUDY OF PLANTS BELONGING TO CAPPARIS

Jamal A.N. Al-Mahweety1image, Ali Alyahawi2image

1School of Chemical Sciences, Faculty of Science, Sana'a University, Yemen

 2Faculty of Medical Sciences, Department of Pharmacy Al-Razi  University, Yemen

ABSTRACT 

Objectives:  Bioactive molecules have gained attention in recent time due to their diverse effects and their economic importance. The objective of current study was to isolate, analyze and identify the active constituents from leaves of C.   cartilaginea. 

Methods:  The powdered leaves were extracted with different solvents by using Soxhlet apparatus and carried out by different chromatographic techniques. The phytochemical characterizations were evaluated by nuclear magnetic resonance and mass spectrometry.

Results:  Three pure compounds were identified representing 1.6% of the total phytochemical composition. Yield of compounds 1, is Kaempferol  (6gm), compound 2, is Dihydroxy-lup-20(29)-en-28-oic acid (3.8gm) and compound 3, is β-Sitosterol (5gm). Three of them are report for first time from this Spp.  

Conclusion:  From this study, can be concluded that report for the first time from C. cartilaginea leaves, identified 3 pure chemical compounds, by analysis using various physical and spectral techniques.

Keywords: Capparis cartilaginea  leaves; Kaempferol, 3,23-Dihydroxy-lup-20(29)-en-28-oic acid, β-Sitosterol,  NMR analysis.


INTRODUCTION

Phytochemicals consists of a various group of natural bioactive molecules very distributed in plants. These bioactive molecules have gained attention in recent times due to their diverse effects on human physiology and human health and their economic importance1. Secondary metabolites are basically classified into three classes namely alkaloids, phenolics and terpenoids2. The molecules in these classes are further grouped due to their numerous modifications, thus emphasizing the diversity of secondary metabolites. Considering this, reasonable attention has been focused on the identification of plants rich in bioactive phytochemicals and characterization of their medicinal and economic values3.

Nutrient limitation, shunt metabolism produced during idiophase, defense mechanism regulator molecules etc,  as a result of secondary metabolism include products an excess metabolism4. Medicinal flora can be use as treatment of diseases and therapeutic agents pro management of health  because they have power over health enhance effects and have bioactive chemical ingredients3. An earlier investigation has reported that Capparis species extracts, such as C. spinosa and C. decidua from Saudi Arabia has a significant anti-inflammatory activity 5 . Newly, the mechanism of anti-inflammatory effect of C. spinosa was suggested by El Azhary et al.,5 and involved inhibition of cellular infiltration and cytokine gene expression. Rutin, a flavonoid, obtained from C. acutifolia Sweet exhibited a potent anti-inflammatory effect6. Other studies have reported the antioxidant, cytotoxic, larvicidal, antimicrobial, hypotensive and bradycardiac activities of C. cartilaginea3,7,8. Capparis cartilaginea have various traditional uses in the Arab region. It is used for easing bruises, childbirth, earache, headache, paralysis, swelling, skin and joint inflammation, knee problems; tendinitis and snakebites5,6. In Yemen, it is called lattssaf, lasaf or nisaf9 and used to treat itching, shortness of breath, head cold, tumors, wounds, boil and for painful knees10,11. Phytochemical studies of C. cartilaginea have resulted in isolating flavonoids (9) and isothiocyanates12. Other phytochemicals, such as carbohydrates, saponins, polyphenols, flavonoids, tannins, triterpenes, sterols, amino acid and protein have also been found in the leaves of C. cartilaginea from Yemen13 . Capers has medicinal distinctive such as rheumatism. Roots used as diuretic, astringent, and tonic14. Bark root used as appetizer, astringent, tonic, antidiarrheic and treat hemorrhoids and spleen disease. Bark was also used for gout and rheumatism, as expectorant, and for chest diseases15. Infusion of stems and root bark were used as antidiarrheic and febrifuge. Fresh fruits were used in sciatica, and dropsy16

 

MATERIALS AND METHODS

TLC and preparative TLC were performed using pre-coated aluminium and glass plates with silica gel 60 F254, whereas column chromatography was carried out on silica gels 70-230 mesh. Spots and bands for compounds on TLC were detected using UV light. Proton NMR (500 MHz) andcarbon-13 NMR (150 MHz) spectra were recorded on JEOL JNM-ECP400 and chemical shifts in ppm were referenced to internal acetone-d6 and CDCl3, respectively.

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Figure 1: Kaempferol

Plant material

collection the plant 26-11-2017 from Al-Mahweet North City in Yemen. Identification and classification of the plant material was performed at the Faculty of Medical Science of the University of Al-Razi specimens were pressed and A voucher specimen (CCJ017) was deposited in a collection housed at the Department of Pharmacy and Pharmacology.

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Figure 2: Dihydroxy-lup-20(29)-en-28-oic acid

Extraction and Isolation

Shade dried leaves of Capparis cartilaginea were powdered (7000gm). Extracted by soxhlet apparatus  with methanol (x4; 3 days). Collective methanolic under vacuum pressure evaporated to yield 8g of the filtrate, then partition with organic solvent (DCM, EtOAc and MeOH) by adding H2O to yield with aqueous methanolic (30 g), dichloromethane (9 g) and ethyl acetate (26 g) partition. 26 g of ethyl acetate fraction chromatographed by using silica gel (70-230 mesh, 1500 g, 9 x 120 cm) C.C., eluting by ethyl acetate and methanol, in order to increase the polarity of solvent and every one of collected partition  was 20ml. jointed fractions (4,11) and rechromatographed by radial chromatography to give CCI (6.00 mg) known as Kaempferol (1), CCII (3.8 mg) known as 3,23-Dihydroxy-lup-20(29)-en-28-oic acid (2) and CCIII (5 mg) known as β-Sitosterol (3). Well-known by using data from earlier NMR and MS spectra.

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Figure 3: Sitosterol

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Figure 4: 1NMR spectra of Sitosterol

Kaempferol (1):- yellowish powder. 1H NMR (CD3COCD3, 500 MHz): δ 8.10 (1H, d, H-2'and6'), 7.80 (1H, d, H-3'and5'),  6.21 (1H, s, H-8), 5.81 (1H, s, H-6). 13C NMR (CD3COCD3, 150 MHz): δ 179.8 (C-4), 166.0 (C-7), 163.1 (C-5), 160.8 (C-4' ), 158.5 (C-2),  156.9 (C-9), 136.0 (C-3), 133.5 (C-2' ),131.9 (6' ), 123.1 (C-1' ), 115.9 (C-3'and5' ), 106.3 (C-10), 101.0 (C-6), 92.9 (C-8).

Dihydroxy-lup-20-en-28-oic acid (2):- white amorphous powder. 1H NMR (CD3COCD3, 500 MHz), δ 4.62, 4,74 (2H, d, H-29), 4.86 (1H, m, H-3), 3.11, 3.45 (2H, s, H-23), 0.79, 0.96, 0.98, 1.03, 1,66 ( each 3H, s, Me×5).  13C NMR (CD3COCD3, 150 MHz): δ 179.7 (C-28), 151.0 (C-20), 112.1 (C-29), 76.0 (C-3), 69.1 (C-23),  53.6 (C-5), 52.0 (C-9), 50.3 (C-18), 48.1 (C-19), 44.1 (C-17), 43.2 (C-14), 41.0 (C-8), 40.0 (C-22), 39.1 (C-13), 38.5 (C-4), 38.0 (C-1), 37.1 (C-10), 37.0 (C-16), 34.6 (C-7), 31.1 (C-21), 28.3 (C-23), 27.4 (C-15), 27.0 (C-12), 25.3 (C-2), 22.1 (C-11), 20.6 (C-30), 19.5 (C-6), 18.1 (C-28), 16.3 (C-25), 16.1 (C-26), 15.5 (C-24), 15.0 (C-27).

Sitosterol (3):- white amorphous powder, 1H NMR (CD3COCD3, 500 MHz): δ 0.68, 0.80, 0.84, 0.91, 1.06 and 1.11 (H- 18,19,21,26,28,29), 5.41 (1H, t, H-6),  3.93 (1H, m, H-3). 13C NMR (CD3COCD3, 150 MHz): δ 141.5 (C-5), 121.1 (C-6), 77.8 (C-3), 57.1 (C-14), 56.2 (C-17), 52.1 (C-24), 50.3 (C-9), 46.1 (C-25), 42.4 (C-13), 41.0 (C-20), 40.2 (C-12), 37.6(C-4), 37.4 (C-1), 36.6 (C-10), 34.0 (C-22), 32.2 (C-8), 32.0 (C-7), 30.4 (C-23),  29.1 (C-16), 28.4 (C-2), 26.1 (C-28), 25.0 (C-15), 21.4 (C-21), 21.3 (C-11), 20.1 (C-27), 19.5 (C-26), 19.0 (C-19), 13.2 (C-29), 12.0 (C-18).

 

RESULTS AND DISCUSSION

The present studies explain the phytochemical investigation of the aerial parts of Capparis  cartilaginea  leaves which resulted in the isolation and structure elucidation of three compounds, hitherto unreported from this part of  plant. The structures of these compounds were elucidated through superior spectroscopic techniques.   

Compound (1);

yellowish powder isolated . IR scale found hydroxyl group at (3395 cm-1) and carbonyl group at (1707 cm-1). The molecular formula is (C16H20O5). The chemical shifts and coupling constants of 1H-NMR data indicated a dihydroxylated pattern for ring A (two m-coupled doublet at δ H 6.21 (1H, s, H-8) and 5.81 (1H, s, H-6) and 4'-  2'and6'), 7.80 (2H, d,  H-3'and5'). Fifteen  carbon signal come out in 13C NMR data as well as six methins and nine quaternary carbons. at δ158.5 and136.0, alkenes carbons group appeared.  

Compound (2); purification as white amorphous powder.  (C30H50O4) is the molecular formula for the compound.  Proton 1H-NMR data exhibit the presence of five tertiary methyl groups at δ H 0.79, 0.96, 0.98, 1.03 and 1.66 for H-(24, 25, 26, 27, 30) positions respectively. The 13C-NMR spectrum of compound display carbonyl of acidic acid, five tertiary methyl groups, terminal double bond and  twenty three signals to other carbon established  in 13C-NMR data. 

Compound (3);  was purified as white amorphous powder. 1H NMR  data appeared at δ (0.68, 0.80, 0.84, 0.91, 1.06 and 1.11), for six methyl groups. Multiple (H-3), Proton appeared at δ (3.93).  Olefinic protons  present at δ 5.41. Spectrum of 13C NMR  showed twenty nine different carbon groups as signal for six methyl's,  nine methylene's, eleven methins and three quaternary carbons. Also appeared at δ141.5 and 121.1, for alkenes carbons. 


Figure 5: 13C spectra for sitoserol

Figure 6: 1NMR spectra of Kaempferol


Figure 7: 13CNMR spectra of Kaempferol

CONCLUSION

Kaempferol, dihydroxy-lup-20(29)-en-28-oic acid  and Sitosterol isolation and purification  from the leaves of  

Capparis  cartilaginea. The  identification and reported for the first time from this part of the plant. The isolation, purification and analysis carried out by means of various physical (solvent extraction, column chromatography, radial chromatography, preparative TLC and malting points) and spectral techniques.

 

AUTHOR’S CONTRIBUTION

The manuscript was carried out, written, and approved in collaboration with all authors.

 

CONFLICT OF INTEREST 

No conflict of interest associated with this work. 

 

REFERENCES

1. Kelechi MN, Nkemakolam AO. Phytochemicals Distribution and Antioxidant Potential of Bauhinia monandra (Linn.) leaves extract. Res J Med Plants 2018; 12 (2): 78-83. 

https://doi.org/10.3923/rjmp.2018.78.83

2. Kennedy DO, Wightman E.L. Herbal extracts and phytochemicals: Plant secondary metabolites and the enhancement of human brain function. Adv Nutr Int Rev J 2011; 2: 32-50. 

https://doi.org/10.3945/an.110.000117

3. Locatelli C, Melucci D, Locatelli M. Toxic metals in herbal medicines- A review. Curr Bioact Compd 2014; 10: 181–188.

https://doi.org/10.2174/1573407210666140716164321

4. The Ayurvedic Pharmacopoeia of India. Part-1 Vol-V, 1st ed., 1999-2000: 41. 

5. El Azhary K, Jouti NT, El Khachibi M, Moutia M, Tabyaoui I, El Hou A, et al. Anti-inflammatory potential of Capparis spinosa L. in vivo in mice through inhibition of cell infiltration and cytokine gene expression. BMC Complement Altern Med 2017; 17:81.

https://doi.org/10.1186/s12906-017-1569-7

6. Chen JJ, Kuo WL, Liao HR, Kuo YH, Chen IS, Shu CW, et al. A new benzenoid and anti-inflammatory constituent of Capparisa cutifolia. Chem Nat Compd 2017; 53:21-3. 

https://doi.org/10.1007/s10600-017-1901-y

7.  Mothana RA, Lindequist U, Gruenert R, Bednarski PJ. Studies of the in vitro anticancer, antimicrobial and antioxidant potentials of selected Yemeni medicinal plants from the island Soqotra. BMC Complement Altern Med 2009; 9:7. https://doi.org/10.1186/1472-6882-9-7

8.  Galib NA, Algfri SK. Phytochemical screening and antioxidant evaluation by DPPH of Capparis cartilaginea decne leaves. J Med Plants 2016; 4:280-6. 

9. Al-Snafi AE. Encyclopedia of the constituents and pharmacological effects of Iraqi medicinal plants. Thiqar University, 2013. 

10. A Guide to Medicinal Plants in North Africa. Asphodelustenui folius Cavan 2005: 70-74.  

11. Esmaeili S, Hamzeloo MM, Ghaffari S, Mosaddegh M. Cytotoxic activity screening of some medicinal plants from south of Iran. Res J Pharmacogn 2014; 1:19-25.  

12.  Latif A, Amer HM, Hamad ME, Alarifi SAR, Almajhdi FN. Medicinal plants from Saudi Arabia and Indonesia: in vitro cytotoxicity evaluation on Vero and HEp-2 cells. J Med Plants Res 2014; 8:1065-1073. 

https://doi.org/10.5897/JMPR2014.5481

13.  Abutaha N, Al-Mekhlafi A. Evaluation of the safe use of the larvicidal fraction of Capparis cartilaginea Decne. against Aedes caspius (Pallas) (Diptera: Culicidae) larvae. Afr Entomol 2014; 22:838-46. 

https://doi.org/10.4001/003.022.0402

14.  Rahimifard N, Shojaii A, Mahbobi M, Hafezan G, Bagheri F, Asgarpanah J. Evaluation of antibacterial activity and flavonoid content of two Capparis species from Iran. J Med Plants 2015;3:89-94.  

15. Bushra AM, Hassan M, Riyadh SA, Faten AAQ.  Phytochemical, anti-inflamatory, anti-oxidant, cytotoxic and anti- bacterial study of Capparis cartilaginea decnefrom Yemen. Int J Pharmacy Pharmaceut Sci 2018; 10, 6: 38-44. 

https://doi.org/10.22159/ijpps.2018v10i6.22905

16. Ibrahim AK, Youssef AI, Arafa AS, Ahmed SA. Anti-H5N1 virus flavonoids from Capparis sinaica Veill. Nat Prod Res 2013; 27(22):2149-2153.

https://doi.org/10.1080/14786419.2013.790027