Phytochemical Investigation and Antioxidant Activity of Hyophorbe Verschaffeltii (Arecaceae)
Total Page:16
File Type:pdf, Size:1020Kb
Journal of Pharmacognosy and Phytochemistry 2016; 5(2): 39-46 E-ISSN: 2278-4136 P-ISSN: 2349-8234 JPP 2016; 5(2): 39-46 Phytochemical investigation and antioxidant activity of Received: 17-01-2016 Accepted: 18-02-2016 Hyophorbe verschaffeltii (Arecaceae) Mohamed R Elgindi A) Department of Mohamed R Elgindi, Abd El-Nassar B Singab, Shaza H Aly, Ibrahim I Pharmacognosy, Faculty of Mahmoud Pharmacy, Egyptian Russian University, Cairo, Egypt. B) Department of Abstract Pharmacognosy, Faculty of The investigation was carried out for isolation and characterization of the possible phytochemical Pharmacy, Helwan University, compounds of leaves of Hyophorbe verschaffeltii and determination of its antioxidant activity. The air Cairo, Egypt. dried leaves of Hyophorbe verschaffeltii were extracted with 70% methanol. The chromatographic investigation for aqueous fraction lead to isolation of five compounds by Column chromatography, thin Abd El-Nassar B Singab layer chromatography (TLC), Preparative thin layer chromatography (PTLC) and paper chromatography. Department of Pharmacognosy, The isolated compounds were identified by spectroscopic techniques as 1H-NMR and 13C-NMR. The Faculty of Pharmacy, Ain Shams 70% methanolic extract was assayed for its antioxidant activity in vivo by CCl4-induced hepatic injury University, Cairo, Egypt. technique and levels of serum liver enzymes Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) were determined, also Oxidative Damage Markers as superoxide dismutase Shaza H Aly (SOD) and malondialdehyde (MDA) in liver tissue were studied. Hyophorbe verschaffeltii (Arecaceae) Department of Pharmacognosy, Faculty of Pharmacy, Badr afforded aqueous fraction from which five compounds Quercetin (compound H-1), Quercetin 7, 3', 4' University in Cairo, Cairo, trimethoxy (compound H-4), Luteolin (compound H-5), Cannigenin (compound H-2) and Brisbagenin Egypt. (compound H-3) were identified for the first time. The H. verschaffeltii extract (200mg/kg) showed a remarkable hepatoprotective and antioxidant activity against CCl4-induced hepatotoxicity as judged from Ibrahim I Mahmoud the serum marker enzymes and antioxidant levels in liver tissues.CCl4-induced a significant rise in ALT, Department of Pharmacognosy, AST, MDA and reduction in SOD level. Treatment of rats with H. verschaffeltii extract (200mg/kg) Faculty of Pharmacy, AlAhram significantly (P<0.001) decrease serum liver enzymes ALT and AST against CCl4-treated rats. Also the Canadian University, Cairo, H. verschaffeltii extract showed significant (P<0.001) elevation of SOD level by 124.7 % as compared to Egypt. control group and the CCl4 intoxicated group treated with H.verschaffeltii showed significant and efficient decline in the level of MDA (P<0.001) compared to CCl4 group by 40.25%. Hyophorbe verschaffeltii have the free radicle scavenging activity that controls the CC4-induced oxidative stress in liver tissue and capable of boosting the intracellular antioxidant capabilities. Keywords: Hyophorbe verschaffeltii, Arecaceae, antioxidant, spirostane, SOD, ALT, Quercetin. Introduction Arecaceae or Palm family (palmeae), in the order Arecales, it is among the famous plant families which include genera that embrace phenolic-rich species, it is a monophyletic group including 183 genera and 2364 species [1, 2]. The palm family (Arecaceae) has a long history of providing man with useful materials for his daily life [3]. Chemically; the family has been neglected despite of its economic importance, probably because of difficulty of collecting fresh material and getting it authenticated. Most work has been carried out on economically important plants such as Phoenix dactylifera, Cocosnucifera and other palms cultivated for their oils [4]. Hyophorbe is a genus of about five species of flowering plants in the Arecaceae family, it contains the following species: Hyophorbe amaricaulis, Hyophorbe indica, [2] Hyophorbe lagenicaulis, Hyophorbe vaughanii and Hyophorbe verschaffeltii . Hyophorbe verschaffeltii is a member of the palm family (Arecaceae, sub-family Arecoideae). This palm is endemic to the Mascarene Islands, which are located to the east of Madagascar in the Indian Ocean [5]. From literature reviews, there is no data reported about neither the phytochemical constituents Correspondence Mohamed R Elgindi nor antioxidant activity of Hyophorbe verschaffeltti H. Wendl. The aim of this study is A) Department of investigation for the chemical constituents and in vivo investigation of antioxidant activity of Pharmacognosy, Faculty of the palm leaves extract. Pharmacy, Egyptian Russian University, Cairo, Egypt. Materials and method B) Department of Pharmacognosy, Faculty of Plant material Pharmacy, Helwan University, Fresh plant material leaves of Hyophorbe verschaffeltti H. Wendl. (Arecaceae) were collected Cairo, Egypt. from El-Zohreya Botanical garden, Cairo, Egypt. The plant was kindly identified by ~ 39 ~ Journal of Pharmacognosy and Phytochemistry Agricultural Engineer Teresa Labib, El Orman Botanical fluorescence under UV-light changed to orange when sprayed Garden. The fresh plant leaves were completely dried in shade with Naturstroff reagent using precoated silica gel TLC. 1H place at room temperature and then powdered by electric mill. NMR (CD3OD, 500MHz): δ ppm 7.8 (1H,d, J =2.1Hz,H-2'), The dried powders were kept in a dark place until subjected to 7.36 (1H,dd, J =2.1, 8.5Hz,H-6'), 6.8 (1H,d,J =8.2Hz, H-5'), the extraction process. 6.42 (1H,d,J =1.8 Hz, H-6), 6.18 (1H,d, J =1.8 Hz, H-8). 13C NMR: δ ppm 178.0 (C-4), 165.2 (C-7), 162.5 (C-5), 158.7(C- Extraction and isolation 9), 148.8 (C-4'), 147.5 (C-2), 145.0 (C-3'), 137.2 (C-3), 124.5 The air dried powdered leaves of Hyophorbe Verschaffeltii (3 (C-1'), 120.8 (C-6'), 116.0 (C-5'), 115.8 (C-2'), 104.4 (C-10), kg) were extracted by maceration with 70% methanol 99.1 (C-6), 94.5 (C-8). (methanol: H2O, 70:30) with occasional stirring at room temperature for 3 days. The process was repeated two times till Compound (H-2) exhaustion. The combined methanol extracts were Crystallized from methanol, TLC investigation using concentrated under reduced pressure at 40º to yield (420g). precoated silica gel G TLC plates and solvent BAW a reddish The residue was suspended in water and diluted with acetone. brown color with spraying reagent 10% H2SO4 with Rf=0.40. 1 The aqueous fraction after filtration was evaporated until H NMR spectral data (500MHz, CD3OD): δ ppm 4.35 dryness to give (41g). About 25 g of the aqueous fraction (1H,m,H-16), 4.01 (1H,m,H-3), 3.78 (1H,m,H-1), 3.48 residue was chromatographed using silica gel column (2H,m,H-26), 0.92 (3H,d,J =7 Hz, H-21), 0.87 (3H,d,J =7 Hz, chromatography (100cm x 2.5cm) using gradient elution H-27), 0.85 (3H,s,H-19), 0.75(3H,s,H-18). 13C NMR: δ ppm system of increasing polarity using chloroform and methanol 109.8 (C-22), 79.9 (C-16), 72.5 (C-1), 65.7 (C-26), 64.6 (C-3), to yield six main fractions (I-VI). Fraction II was eluted with 62.2 (C-17), 55.9 (C-14), 54.7 (C-9), 41 (C-13), 39.5 (C-2), chloroform-methanol (90:10) revealed the presence of yellow 37.3 (C-5), 36.2 (C-4), 35.3 (C-8), 31.8 (C-15 & C-23), 30.8 fluorescent spot with Rf = 0.95 using silica gel GF254 plates and (C-25), 29.5 (C-7), 28.5 (C-24), 27 (C-6), 23.5 (C-11), 14.1 BAW then subjected to Co-chromatograms using Whatmann (C-21), 16.6 (C-27). No.1 MM paper chromatography using 2 solvent systems: BAW and 15% AcOH, these were examined under UV light Compound (H-3) before and after exposure to ammonia vapors then purified Crystallized from methanol, TLC investigation using over preparative TLC and concentrated under reduced pressure precoated silica gel G TLC plates and solvent BAW a reddish to yield compound H-1 (40 mg). Fraction IV eluted with brown color with spraying reagent 10% H2SO4 with Rf =0.415. 1 chloroform-methanol (70:30) revealed the presence of major H NMR spectral data (500MHz, CD3OD): δ ppm 4.18 (1H, green color spot with Rf = 0.737 after spraying with 10% v/v m,H-16), 3.99 (1H,dd, J =11.3, 4.6 Hz,H-3), 3.73 (1H,dd, J sulphuric acid followed by heating at 100º for 10 minutes, =11.3, 4.4 Hz, H-1), 3.63 (1H, dd, J =10.5, 3.5Hz, H-26ax.), using silica gel GF254 plates and BAW. These fractions were 3.36 (1H, dd, J =10.5, 10.5Hz, H-26eq.), 0.96 (3H,d, J =7.0 combined together and concentrated under reduced pressure to Hz, H-21), 0.88 (3H,s, H-19), 0.85 (3H,d, J =5.6 Hz, H-27). afford a residue of group IV. This residue was subjected for 13C NMR: δ ppm 108.9 (C-22), 81.8 (C-16), 77 (C-1), 67.8 (C- further purification by another silica gel sub-column 3), 66.2 (C-26), 62.9 (C-17), 59.15 (C-14), 54.8 (C-9), 44.1 chromatography (66g, 75x2 cm) using (75:25 CHCl3: MeOH) (C-2), 42.9 (C-5), 42.5 (C-20), 42.1 (C-10), 41.1 (C-12), 40.5 solvent system then gradient elution till 100% MeOH, Sub- (C-13), 39(C-4), 36.3 (C-25), 36 (C-8), 33.3 (C-15), 32.3 (C- fraction IV-I was purified over preparative TLC and 23), 32.2 (C-7), 29.9 (C-24), 29.7 (C-6), 24.5 (C-11), 18 (C- concentrated to yield compound H-2(25mg), and sub-fraction 27), 16.8 (C-18), 13.1 (C-21), 6.8 (C-19).