INTRODUCTION:

“To take medicine only when you are sick is like digging a well only when you are thirsty — is it not already too late?” (Chi Po, c 2500 BC). This proverb suggests that prevention is more important than treatment [1, 2]. Plants have always been usable sources of drugs, and many currently available drugs are directly or indirectly derived from plants. Many of the oral agents that are presently in use for the treatment of “life style diseases” suffer from implication in a number of serious and adverse effects [3]. Therefore, it is important to investigate the biologically active components of plants with hypoglycemic actions which include flavonoids, alkaloids, glycosides, polysaccharides, and peptidoglycan [4, 5].

The “functional food” industry has produced and marketed foods enriched with bioactive compounds, but there are no universally accepted criteria for judging efficacy of the compounds or enriched foods. The lack of understanding on bioactive compounds and their health benefits should not serve to reduce research interest but should instead encourage plant and nutritional scientists to work together to develop strategies for improvement of health through food [6].

An edible and medicinal tropic plant—Morinda citrifolia L (Noni)

The ancestors of Polynesians are believed to have brought many plants with them, as food and medicine, when they migrated from Southeast Asia 2000 years ago[7]. Of the 12 most common medicinal plants they brought, Noni was the second most popular plant used in herbal remedies to treat various common diseases and to maintain overall good health[8]. It bears botanical name Morinda citrifolia possesses several chemical and biological properties. Morinda citrifolia, commonly known as Great Morinda, Indian mulberry, Nunaakai (Tamil Nadu, India), dog dumpling (Barbados), Mengkudu (Indonesia and Malaysia), Kumudu (Balinese), Pace (Javanese), Beach mulberry, Cheese fruit[9] or Noni (from Hawaiian) is a tree in the coffee family, Rubiaceae. Morinda citrifolia native range extends through Southeast Asia (Malaysia) and Australia, and the species is now cultivated throughout the tropics and widely naturalized.[10] It has been reported to have a broad range of health benefits for cancer, infection, arthritis, diabetes, asthma, hypertension, and pain[11]. The Polynesians utilized the whole Noni plant in their medicinal remedies and dye for some of their traditional clothes. The roots, stems, bark, leaves, flowers, and fruits of the Noni plant are all involved in various combinations in almost 40 known and recorded herbal remedies [12]. Additionally, the roots were used to produce a yellow or red dye for tapa cloths and fala (mats), while the fruit was eaten for health and food (figure 3 A, B, C, and D).

There are numerous Polynesian stories of heroes and heroines that used Noni to survive from famine. There is one tale of Kamapua’a, the ancient god, who loved Pele, the volcano goddess. He taunted Pele with a chant, “I have seen the woman gathering Noni /scratching Noni/pounding Noni.”

Supposedly, the chant referred to Pele’s eyes becoming red, and she became so angry that she plunged into battle with him. A Tongan myth tells of the God Maui being restored to life by having Noni leaves placed on his body [13].

Isabel Abbott, a former botanical chemist at the University of Hawaii, stated that, “People are crazy about this plant. They use it for diabetes, high blood pressure, cancer, and many other illnesses” [14]. Bushnell reported that Noni was a traditional remedy used to treat broken bones, deep cuts, bruises, sores, and wounds [15]. Morton gave numerous references for medicinal uses of Noni [16]. Joseph Betz, a research chemist in the FDA’s Division of Natural Products, Center for Food Safety and Applied Nutrition, stated that “Morinda citrifolia has been tested for a number of biological activities in animal and anti-microbial studies.” He reports that the dried fruit has smooth muscle stimulatory activity and histaminergic effects [17].

Nutrients and Phytochemicals of Mengkudu [18–37]

Micheal Tierra is a well respected herbalist who practices Western, European and Chinese traditional medicine. In his book “the way of herbs” he lists the healing properties that are contained in the biochemical constituents of Noni. This list of Noni’s properties clearly illustrates that it is “pro” everything good and “anti” everything bad. The partial lists include antimicrobial, anti-inflammatory, antiarthritic, antihypertensive, antiulcer, anticancer, and antioxidants effects.

A number of foremost components have been identified in the Noni plant such as scopoletin, octanoic acid, potassium, vitamin C, terpenoids, alkaloids, anthraquinones (such as nordamnacanthal, morindone, rubiadin, and rubiadin-1-methyl ether, anthraquinone glycoside), β-sitosterol, carotene, vitamin A, flavone glycosides, linoleic acid, alizarin, amino acids, acubin, L-asperuloside, caproic acid, caprylic acid, ursolic acid, rutin, and a putative proxeronine. These constituents and their classes are listed in Table (1) and references therein. The recently reported biological effects of Mengkudu are tabulated along with respective references in Table 2.

Scopoletin [38, 39]

Recently, Duncan demonstrated that scopoletin, a health promoter coumarin in Noni, inhibits the activity of E coli, commonly associated with recent outbreaks resulting in hundreds of serious infections and even death. Noni also helps stomach ulcer through inhibition of the bacteria H. pylori.

Screening test for coumarin [40]

About 3 g of the test sample was dissolved in distilled water in the flask. The flask was first covered with a piece of filter paper moistened with 1 M sodium hydroxide solution and firmly with a piece to aluminum foil on the top. The solution was then warmed on the water bath for a few minutes. Finally the filter paper was examined under the UV-light. The appearance of fluorescence after a short time indicated the presence of coumarin or related volatile compounds in the test sample.

Isolation [41–43]

The fruits of Mengkudu (8 kg) were extracted four times with methanol (MeOH) at room temperature (r.t.). The solvent was removed under reduced pressure to yield 533.0 g of a thick syrupy extract. The extract was fractionated with H₂0 and ethyl acetate (EtOAc) to yield an EtOAc phase and the main aqueous phase. The EtOAc phase was dried (with anhydrous Na2SO4), and the solvent was evaporated under vacuum. The residue (33.5 g) from the EtOAc extract was then fractionated with n-hexane to produce n-hexane soluble and -insoluble fractions. The latter fraction was subsequently treated with Et20 to yield Et₂0-soluble and Et₂0-insoluble fractions. The Et₂0-insoluble fraction was further divided into EtOAc-soluble and EtOAc-insoluble fractions. The EtOAc-soluble fraction was subjected to vacuum liquid chromatography (VLC; silica gel PF2~, n hexane, n-hexane-EtOAc, CHCI₃, CHCI₃-MeOH, MeOH) to yield 13 fractions. Of these, Fr. 5 (2 g) was further separated by column chromatography (CC; silica gel PF2S₄, CHCI₃, CHCI₃-MeOH, MeOH). As a result, 18 fractions were obtained and combined on the basis of TLC to ultimately yield six fractions (A'-F'). Fr. C' (200 mg) was again subjected to CC (n-hexane, n-hexane-EtOAc, CHCI₃, CHCI₃-MeOH and MeOH in order of increasing polarity). From this separation, a total of nine fractions (C1'-C9') were obtained by combining various fractions based on TLC results. Fr. D' (100 mg) from the original CC was again subjected to CC (n-hexane, n-hexane-EtOAc, CHCI₃, CHCI₃-MeOH, and MeOH) resulting in seven fractions (DI'-D7'). Of these, Fr. D4' was purified by prep. TLC (CHCI3-MeOH; 9:1) to furnish scopoletin compound as white needles (8 mg).

Synthesis of scopoletin [44 – 46]

Scopoletin (6-methoxy-7-hydroxycoumarin) was synthesized starting from commercial isovanillin by Crosby (1961) with 30% overall yield. In 1962, Crosby and Berthold developed a one-pot synthesis of scopoletin with 73% yield. In 2002, Demytteneare et al. reported the synthesis of 6-methoxy benzopyran-7-ol via scopoletin as an intermediate. Last but not least method of synthesis is as follows: 2, 4-dihydroxy-5-methoxybenzaldehyde was obtained in a one-step reaction from 2, 4, 5-trimethoxybenzaldehyde by reaction with aluminium (III) chloride in dichloromethane, followed by acid hydrolysis. Treatment with malonic acid in pyridine for 24 h at room temperature (r.t) using phenylamine as catalysts afforded 7-hydroxy-6-methoxy-2- oxo-2H-chromene-3-carboxylic acid in 86% yield. Then heating 7-hydroxy-6-methoxy-2-oxo-2H-chromene-3-carboxylic acid in a pyridine/ethylene glycol mixture (1:1.1) to reflux for 3 h gave scopoletin (figure 3 E).

Biological functions of scopoletin [47 – 50]

Scopoletin is one of the main coumarin constituents that occur in the fruits of Mengkudu. It possesses a wide range of biological activities such as anti-inflammatory, hypouricemic, and antioxidant activities. Reports have indicated that several coumarin-type compounds block angiogenesis by inhibiting the endothelial cell growth. Previous data also demonstrated that scopoletin and its derivatives have anti-tumor and anti-angiogenic activities. These findings pave the way for future studies on the anti-angiogenic potential and related mechanisms of scopoletin derivatives.

Purity assessment and Quantification of Scopoletin [51– 56]

Chemical marker standard scopoletin was isolated directly from the freeze-dried Noni fruit and leaf powder. Identity and purity (>98%) were confirmed by NMR, MS, and HPLC. The marker compound was dissolved in methanol (MeOH) to a concentration of 1 mg/mL. Eight raw Noni fruit samples (F1-F8) were collected from different locations including French Polynesia (Tahiti, Moorea, and Motu Fareone), Tonga, Dominican Republic, Okinawa, Thailand, and Hawaii. The fruit samples were stored below 0ºC before use. Fruits were thawed, mashed and extracted (2 g) twice with 125 mL methanol (MeOH), aided by sonication for 30 min. The solvent was removed under vacuum in a rotary evaporator. The MeOH extracts w ere then redissolved in 10 mL of MeOH. Commercial Noni fruit juice products (J1-J4), originating from Tahiti, Dominican Republic, Hawaii, and Costa Rica, produced by different manufacturers, were purchased at local markets or via the internet. Prior to analysis, all samples were filtered through a 0.45 m nylon membrane filter and then purified by Solid-Phase Extraction (SPE) with Waters OASISS® extraction cartridges. SPE cartridges were first equilibrated with water, followed by methanol. The samples were then loaded onto the cartridge and washed with 5% MeOH, followed by 100% MeOH. The MeOH eluate was retained for TLC analysis. Four different Noni fruit powder capsule products were also purchase. The producers of the capsules are located in French Polynesia (C1), Hainan, South China Sea (C2), Hawaii (C3), and Indonesia (C4). One gram of the capsule contents was extracted with 5 mL MeOH, aided by sonication for 10 min. The MeOH extracts were filtered, and the solvent removed by evaporation under vacuum at 50ºC. The extract was then redissolved in 1 mL of MeOH.

Analysis of scopoletin content of the samples was also performed by HPLC, according to a previously reported method. Chemical standards of scopoletin was accurately weighed and then dissolved in an appropriate volume of MeOH/MeCN to produce corresponding stock standard solutions. Working standard solutions for calibration curve was prepared by diluting the stock solutions with MeOH at different concentrations. All stock and working solutions were maintained at 0ºC in a refrigerator. Samples were extracted and dissolved in MeOH. Chromatographic separation was performed on a Waters 2690 separations module coupled with 996 a photodiode array (PDA) detector, and equipped with a C18 column. The mobile phase system was composed of three solvents: A; MeCN, B; MeOH and C; 0.1 % TFA in H2O (v/v). The mobile phase was programmed consecutively in linear gradients as follows: 0 min, 10% A, 10% B, and 80% C; 15 min, 20% A, 20% B, and 60% C; 26 min, 40% A, 40% B, and 20% C; 28–39 min, 50% A, 50% B, and 0% C; and 40-45 min, 10% A, 10% B, and 80% C. The elution was run at a flow rate of 1.0 mL/min. The UV spectra were quantified at 365 nm.

Table 3: TLC method parameters for chemical marker scopoletin of Noni fruit

Chemical marker

Mobile phase

characteristic

Visualization method

Spot

Scopoletin

Dichloromethane:

methanol

(19: 1, v:v)

UV lamp, 365 nm

Fluorescent light blue spot

Fig 1 .Scopoletin analysis of commercial Noni juice (J1-J4) and Noni capsules (C1-C4) by TLC compared with scopoletin (S).

Fig. 2: Scopoletin analysis of Noni fruit F1-F8 by TLC compared with scopoletin (S).

Figure 3 A. illustrates the Mengkudu tree, B. Fruit with syncarpous flower, C. Ripe fruit and D. Fruit with seed and E. Chemical structure of scopoletin.

Results of the HPLC analyses indicate a wide range in the concentrations of marker compounds among the various sources of Noni fruit and leaves, and resulting commercial products. The analyses confirmed the presence of scopoletin in all Noni fruit based samples. Scopoletin was present in the range of 0.7-6.9 mg/g. Noni fruit powder capsules contained 0.1-0.4 mg scopoletin/g. Scopoletin occurred in commercial Noni juice products at 3.7-21.2 µg/mL.

In the scopoletin analyses, fluorescent light blue spots were visible to the unaided eye on the developed TLC plates, when viewed under long wave UV light, 365 nm (Table 3). The scopoletin standard produced the most intensely fluorescent light blue spot. Every Noni fruit, juice, and capsule also produced the same spot with a retention factor (Rf) of 0.5, but with differing intensities (Fig. 1 and 2). Scopoletin was also detected by this TLC method at the lowest concentration in any sample, which was 3.7 g/mL found in sample J3 (Fig. 1). At this low concentration, a faintly fluorescent light blue spot was still visible.

The TLC analysis results were confirmed by the result of the HPLC analyses. The sensitivity of the scopoletin TLC method is apparently very good, since as little as 3.7 g/mL produced a fluorescent blue spot that was discernable to the unaided eye. Quantities lower than these are possible in some commercial products where Noni ingredients are blended with others. However, detection in these products can be enhanced by concentration of the extracts produced during sample preparation.

The utility of the TLC methods for the identification of Noni fruit ingredients, as well as Noni based commercial products, has been demonstrated. These methods do not require expensive instrumentation or specialized laboratories. Therefore, the methods are readily transferable to analysts working under a variety of circumstances. The marker compounds utilized in these methods are characteristic of Noni fruit from all regions where this plant is cultivated.

Pharmacokinetic study of scopoletin [57]

The pharmacokinetics of Noni was studied in female SD rats after oral administration at a dose of 1 mL Noni puree per 100 g body weight. A known major component (scopoletin) in Noni was chosen as a marker and monitored in the plasma and in different organs over time by HPLC analysis in the cooperation with Rand D, Department of Morinda, Inc. The pharmacokinetics of scopoletin in Noni puree was calculated as follows: the plasma concentration reached a peak at 2 h after oral administration of Noni. The peak level of scopoletin decreased to 50 % in 4 h. Only 12 % and 2 % of the scopoletin was left in the plasma at 12 and 24 h, respectively. Absorption was rapid, with 50 % peak concentration reached in only 30 min. In order to maintain a higher blood level of scopoletin, TNJ should be taken every 2 to 4 h. For overall health maintenance, TNJ should be taken in one-ounce servings every 12 h. The results demonstrate that the frequency of drinking TNJ is more important than the amount. The concentration of scopoletin in various organs indicates that Noni is absorbed into different tissues approximately one hour after administration. The peak concentration in different tissues occurred at about 3 h, with a rapid decline. Interestingly, the scopoletin level in breast tissue was relatively higher than any other extra-GI tract tissue.

Scopoletin a brief insight at molecular level

Scopoletin besides its molecular circumcision’s needs to be defined as if to be used for further research and biomedical and therapeutic use at anti proliferative, anti-diabetic and anti-microbial events at molecular and cellular levels. Recently, it has been found that the scopoletin significantly inhibited phorbol myristate acetate (PMA)/ionomycin-induced interleukin-4 (IL-4), IL-5, and IL-10 production in EL-4 T cells and it also significantly enhanced the inhibitory action of PMA/ionomycin on interferon-γ (IFN-γ) expression [58]. Cheng and colleagues also showed that in EL-4 T cells, PMA/ionomycin treatment markedly increased the expression of nuclear factor of activated T cells (NFAT) and GATA-3; in contrast, scopoletin significantly down-regulated expressions of these transcription factors. It has been suggested that this down regulation depended on protein kinase C (PKC) attenuation. This leads us to suggest that the anti-allergic properties of scopoletin might be mediated by the down regulation of cytokine expression in Th 2 cells. Nevertheless, Moon and colleagues [59] studied the effect of scopoletin in human mast cell line (HMC-1) and they reported significant dose-dependent level of scopoletin inhibits the way, in which phorbol 12-myristate 13-acetate (PMA) plus A23187 induces the production of inflammatory cytokines such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-8 (P<0.05). The maximal rates at which scopoletin (0.2 mM) inhibited the production of TNF-alpha, IL-6, and IL -8 were 41.6%+/-4.2%, 71.9%+/-2.5%, and 43.0%+/-5.7%, respectively. In activated HMC-1 cells, the expression level of nuclear factor (NF)-kappaB/Rel A protein was found to be increased in the nucleus whereas the level of NF-kappaB/Rel A in nucleus was decreased by treatment with scopoletin. Scopoletin decreased PMA plus A23187-induced luciferase activity. Scopoletin also inhibits IkappaBalpha phosphorylation and degradation in cytoplasm. It was reported that the scopoletin has a potential regulatory effect on inflammatory reactions that are mediated by mast cells [59].

Beside anti-inflammatory effect, scopoletin has been reported to have a hypotensive effect. Ojewole and colleagues [60] showed that scopoletin inhibits the indirect electrical stimulation-evoked contractions of the cat nictitating membrane and also the contractions of isolated perfused central ear artery of rabbit, induced by electrical stimulation or intraluminal noradrenaline administration. This coumarin, like papaverine, reduces the amplitude and frequency of the spontaneous, myogenic, rhythmic contractions, and exogenous noradrenaline-evoked contractions of the rat isolated portal vein. Scopoletin also inhibits the spontaneous, myogenic, pendular, rhythmic contractions of the rabbit isolated duodenum and attenuates the indirect electrical stimulation-provoked or exogenous noradrenaline-induced relaxations of the muscle preparation. It also depresses the electrical stimulation-evoked contractions of the chick isolated oesophagus. Scopoletin, on its own accord, relaxes all the smooth muscles examined and inhibits the spasmogenic activities of a wide variety of agonists on guinea-pig isolated ileum to approximately the same extent. It is therefore speculated that scopoletin probably produces hypotension in laboratory animals through (a) its smooth muscle relaxant activity; by which means it presumably dilates blood vessels; and (b) by acting as a non-specific spasmolytic agent (like papaverine) [60].

To cap it all, based on the literature, exceptionally pertaining to traditional literature and studies carried out in a scientific manner, scopoletin from Morinda citrifolia can be considered to be secondary metabolite (coumarin) with great potential for resolving life style oriented health problems. This is in light of the fact that some of the major diseases that plague mankind revolve around the inability of the system to respond to oxidative stress and the subsequent decrease in the antioxidant status that creates more deleterious effects in the system. It is learnt through literature that scopoletin have much more ability to restore the concentration of the antioxidant status when compared to crude fruit extract. Hence, it is hypothesized that the scopoletin is exhibiting commendable clinical efficacy. It can be recommended that scopoletin extracts are potent candidates for further clinical studies and can possibly end the elusive search for good therapeutic agents.

ACKNOWLEDGEMENT:

This review is part of the FRGS project entitled ‘Extraction, bioactivity guided fractionation and characterization of bioactive principles from Morinda citrifolia (Mengkudu) fruits for the treatment of diabetes’.

REFERENCES:

1. Veith I. Translated “Yellow Emperor’s Classic of Internal Medicine” (2500 BC); 2002.

2. Du B, You S. Present situation in preventing and treating liver fibrosis with TCM drugs. J Tradit Chin Med. 2001 Jun; 21(2):147-52.

3. Zhang BB, Moller DE (2000) New approaches in the treatment of type 2 diabetes. Curr Opin Chem Biol 4:461–467.

4. Grover JK, Vats V, Yadav SP (2002) Effect of feeding aqueous extract of Pterocarpus marsupium on glycogen content of tissues and the key enzymes of carbohydrate metabolism. Mol Cell Biochem 241:53–59

5. Mao CP, Xie ML, Gu ZL (2002) Effects of konjac extract on insulin sensitivity in high fat diet rats. Acta Pharmacol Sin 23:855–859

6. Abhang R Y. Jogiekar P P and Kulkarni P H, Preliminary study on the effect of T.cordifolia ,on mitosis, Ancient Sci, (1991)1.27.

7. Plants by Common Name — James Cook University

8. Nelson, SC (2006-04-01). "Species Profiles for Pacific Island Agroforestry: Morinda citrifolia (Noni)". http://traditionaltree.org.

9. Tabrah FL, Eveleth BM. Evaluation of the effectiveness of ancient Hawaiian medicine. HawaiiMed J 1966; 25: 223-30.

10. Krauss B. Plants in Hawaiian culture. Honolulu: University of Hawaii Press; 1993. p103, p252.

11. WhistlerW. Tongan herbalmedicine. Isle Botanica,Honolulu, Hawaii, 1992. p 89-90.

12. Bruggnecate JT. Native plants can heal your wounds. Honolulu Star-Bulletin Local News 1992 Feb 2.

13. NealM. Gardens ofHawaii. Honolulu, Hawaii: Bishop Museum Press; 1965. p 804.

14. Abbott IA. The geographic origin of the plants most commonly used for medicine by Hawaiians. J Ethnopharmacol 1985; 14: 213-22.

15. Bushnell OA, Fukuda M, Makinodian T. The antibacterial properties of some plants found in Hawaii. Pacific Science 1950; 4: 167-83.

16. Morton JF. The ocean-going Noni, or Indian mulberry (Morinda citrifolia, Rubiaceae) and some of its ‘colorful’ relatives. Economic Botany 1992; 46: 241-56.

17. Pride Publishing,Noni: Polynesia’s natural pharmacy. 1997. p 13.

18. Wang MY, West BJ, Jensen CJ, Nowicki D, Su C, Palu AK, Anderson G (2002) Morinda citrifolia (Noni): a literature review and recent advances in Noni research. Acta Pharmacol Sin 23:1127-1141.

19. Hirazumi A,Furusawa E. 1999.Animmunomodulatory polysaccharide-rich substance from the fruit juice of Morinda citrifolia (Noni) with antitumour activity. Phytother Res 13: 380–7.

20. Wang, Y. Jin, N. Nakatani, N. Zhu, K. Csiszar, C. Boyd, R. T. Rosen, G. Ghai and C. Ho. 2000. Novel glycosides from Noni (Morinda citrifolia). J. Nat. Prod. 63: 1182-1183.

21. Sang, S., X. Cheng, N. Zhu, M. Wang, J. Jhoo, R. Stark, V. Badmaev, G. Ghai, R. T. Rosen and C. Ho. 2001. Iridoid glycosides from the leaves of Morinda citrifolia. J. Nat. Prod. 64: 799-800.

22. Farine JP, Legal L, Moreteau B, Le Quere JL. Volatile components of ripe fruits of Mor inda citr ifolia and their effects on Drosophila. Phytochemistry 1996; 41: 433-8.

23. Dyas, A., R. Wise and J. Pijck, 1983. Reproducibility study of pharmacokinetics of amikacin, gentamicin and tobramycin: A three way cross over study. Journal of Antimicrobial Chemotherapy, 12, 371−376.

24. Makishima, K.; Ohashi, T.; Inoue, H.; Koyama, K.; Matsuoka, M.; Murakami, T.; Oda, M.; Ogawara, Y. et al. (1981). "Discovery of two new burst sources in the globular clusters Terzan 1 and Terzan 5". The Astrophysical Journal 247: L23–25.

25. Wang, Z., Wilson, G.F., Griffith, L.C. (2002). Calcium/calmodulin-dependent protein kinase II phosphorylates and regulates the Drosophila Eag potassium channel. J. Biol. Chem. 277(27): 24022--24029.

26. Duncan, D. K., Peterson, R. C., Thorburn, J. A., and Pinsonneault, M. H. 1998, ApJ, 499, 2

27. Hirazumi, A., Furusawa, E., Chou, S.C., Hokama, Y., 1994. Anticancer activity of Morinda citrifolia (Noni) on intraperitoneally implanted Lewis lung carcinoma in syngeneic mice. In: Proceedings of the Western Pharmacology Society 37, pp. 145–146.

28. Furusawa K, Tajima F, Umezu Y, Ueta M, Ide M, Mizushima T and Ogata H (2003). Activation of natural killer cell functions in recreational athletes with paraplegia during a wheelchair half-marathon race. Arch Phys Med Rehabil 84:706-711.

29. Hiramatsu, O., Goto, M., Yada, T., Kimura, A., Tachibana, H., Ogasawara, Y., Tsujioka, K. and Kajiya, F. (1994). Diameters of subendocardial arterioles and venules during prolonged diastole in canine left ventricles. Circulation Research 75, 393—399.

30. Hiwasa, T., Arase, Y., Chen, Z., Kita, K., Umezawa, K., Ito, H. and Suzuki, N. 1999. Stimulation of ultraviolet-induced apoptosis of human fibroblast UV-1 cells by tyrosine kinase inhibitors. FEBS Letters 444: 173-176.

31. Liu, G, J. A. Curry, J. A. Haggerty, and Y. Fu, Retrieval and characterization of cloud liquid water path using airborne passive microwave data during INDOEX, J. Geophys. Res., 106, 28,719– 28,730, 2001.

32. Sang Y, Cui D, Wang X (2001) Phospholipase D and phosphatidic acidmediated generation of superoxide in Arabidopsis. Plant Physiol 126: 1449–1458

33. M.S. Hounzangbe-Adotea, V. Paolinib, I. Fourastec, K. Moutairoua, H. Hoste .In vitro effects of four tropical plants on three life-cycle stages of the parasitic nematode, Haemonchus contortus. Research in Veterinary Science Volume 78, Issue 2, April 2005, Pages 155–160.

34. Y. V. Li, C. J. Hough, J. M. Sarvey, Do We Need Zinc to Think? Sci. STKE 2003, pe19 (2003)

35. Kamiya, K. et al. Chemical constituents of Morinda citrifolia fruits inhibit copper-induced low-density lipoprotein oxidation. Journal of Agricultural and Food Chemistry, v. 52, p. 5843–5848, 2004.

36. Hornick, C. A. et al. Inhibition of angiogenic initiation and disruption of newly established human vascular networks by juice from Morinda citrifolia (Noni). Angiogenesis, v. 6, p. 143–149, 2003.

37. Hirazumi, A., Furusawa, E., Chou, S.C., Hokama, Y., 1996. Immunomodulation contributes to the anticancer activity of Morinda citrifolia (Noni) fruit juice. In: Proceedings of the Western Pharmacology Society 39, pp. 7–9.

38. Duncan SH, Flint HJ, Stewart CS. Inhibitory activity of gut bacteria against Es cherichia coli 0157 mediated by dietary plant metabolites. FEMS Microbiol Lett 1998; 164: 283-58.

39. Mahattanadula, S., Ridtitid, W., Nima, S., Phdoongsombutc, N., Ratanasuwond, P., and Kasiwonga, S. (2011). Effects of Morinda citrifolia aqueous fruit extract and its biomarker scopoletin on reflux esophagitis and gastric ulcer in rats. Journal of Ethnopharmacology, 134, 243–250.

40. Rizk, A.M., 1982. Constituents of plants growing in Qatar. Fitoterapia, 52: 35-42.

41. Iwagawa, T. and Hase, T., A coumarin acetylglucoside from Vibumum suspen-sum. Phytochemist~ 23,467-468 (1984).

42. Mulder-Krieger, T., Verpoorte, R., Water, A., Gessel, M. V., Oeveren, B. C. J. A. V., and Svendsen, A. B., Identification of the alkaloids and anthraquinones in Cinchona ledgeriana callus cultures. Planta Med., 46, 19-24 (1982).

43. Hill, R. A., Krebs, H. C., Verpoorte, R., and Wijnsma, R., Progress in the Chemistry of Organic Natural Products, Springer- Verlag Wien New York, vol. 49, pp. 79-149 (1986).

44. Braymer, H. D., M. R. Shetlar and S. H. Wender. 1960. An improved synthesis of scopoletin. Biochim. Biophys. Acta. 44: 163-164.

45. Crosby, D. G. 1961. Improved synthesis of scopoletin. J. Org. Chem. 26: 1215-1217.

46. Crosby, D. G. and R. V. Berthold. 1962. New syntheses in the coumarin series. J. Org. Chem. 27: 3083-3085.

47. Moon, P. D.; Lee, B. H.; Jeong, H. J.; An, H. J.; Park, S. J.; Kim, H. R.; Ko, S. G.; Um, J. Y.; Hong, S. H.; Kim, H. M. Eur. J. Pharmacol. 2007, 555, 218.

48. Nam, N. H.; Kim, Y.; You, Y. J.; Hong, D. H.; Kim, H. M.; Ahn, B. Z. Bioorg. Med. Chem. Lett. 2002, 12, 2345.

49. Lee, S.; Sivakumar, K.; Shin, W. S.; Xie, F.; Wang, Q. Bioorg. Med. Chem. Lett. 2006, 16, 4596.

50. Zhou, J.; Wang, L.; Wei, L.; Zheng, Y.; Zhang, H.; Wang, Y.; Cao, P.; Niu, A.; Wang, J.; Dai, Y. Lett. Drug Des. Disc. 2012, 9, 397.

51. Deng, S., B. West, A. Palu and J. Jensen, Determination and comparative analysis of major iridoids in different parts and cultivation sources of Morinda citrifolia. Phytochem. Anal., 2011; 22:26–30.

52. Deng, S., B.J. West and C.J. Jensen, Simultaneous characterization and quantitation of flavonol glycosides and aglycones in Noni leaves using a validated HPLC-UV/MS method. Food Chem., 2007; 111(2): 526-29.

53. Deng, S., B.J. West and J. Jensen,. A quantitative comparison of phytochemical components in global Noni fruits and their commercial products. Food Chem., 2010; 122(1): 267-70.

54. Harborne, J.B., 1998. Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. Chapman and Hall, London.

55. Inouye, H., Y. Takeda, H. Nishimura, A. Kanomi, T. Okuda and C. Puff, 1988. Studies on monoterpene glucosides and related natural products. Part 61.Chemotaxonomic studies of Rubiaceous plants containing iridoid glycosides. Phytochemistry, 27: 2591-8.

56. Kamiya, K., Y. Tanaka, H. Endang, M. Umar and T. Satake, 2005. New anthraquinone and iridoid from the fruits of Morinda citrifolia. Chem. Pharm. Bull., 53: 1597-1599.

57. WANG Mian-Ying2, Brett J WEST3, C Jarakae JENSEN3, Diane NOWICKI, SU Chen3, Afa K PALU3, Gary ANDERSON, Morinda citrifolia (Noni): A literature review and recent advances in Noni research, Acta Pharmacol Sin., 2002; 23 (12): 1127-41.

58. Cheng AS, Cheng YH, Chang TL. Scopoletin attenuates allergy by inhibiting Th2 cytokines production in EL-4 T cells. Food Funct. 2012 Aug;3(8):886-90.

59. Moon PD, Lee BH, Jeong HJ, An HJ, Park SJ, Kim HR, Ko SG, Um JY, Hong SH, Kim HM. Use of scopoletin to inhibit the production of inflammatory cytokines through inhibition of the IkappaB/NF-kappaB signal cascade in the human mast cell line HMC-1. Eur J Pharmacol. 2007 Jan 26;555(2-3):218-25.

60. Ojewole JA, Adesina SK. Mechanism of the Hypotensive Effect of Scopoletin Isolated from the Fruit of Tetrapleura tetraptera. Planta Med. 1983 Sep;49(9):46-50.

Received on 02.07.2013 Modified on 25.07.2013

Research J. Pharm. and Tech. 6(9): September 2013; Page 978-984

Classes	Compounds	Occurrence
Anthraquinones	Morindine, rubiadine Rubiadine 1- methylether	Roots and fruit
Glycosides	Glycoside of coumarin,flavone and anthraquinone	Fruit
Essential oils	Volatile oil	Ripe fruit
Coumarin	Scopoletin	Fruit
Flavonol	Vomifoliol	Ripe fruit
Monoterpenes	Iridoid	Leaves
Sterol	Campesterol, Stigmasterol, Sitosterol,Isofucosterol, Sitosteryl palmitate, Isofucosteryl palmitate	Cell suspension culture of M. citrifolia
Vitamins	Vitamin C 24- 258 mg/100 g dried fruit	Dried fruit
Anthraquinones	Rubiadin lucidin, morindone, lucidin-3- prineresal, morindone-6- β –primeveroside, seven new quinones	Cell suspension culture of M. citrifolia

Biological Effects	References
Antibacterial activity	25
A health promoter that inhibits the activity of E. coli; also helps in stomach ulcer treatment through inhibition of the H. pylori bacteria.	26
Suppression of cytopathic effect of HIV infected MT-4 cells, without inhibiting cell growth.	25
Mycobacterium tuberculosis killer in in vitro study	25
Anticancer activity	27, 28
Enhancement of the therapeutic effect of anticancer drugs such as Taxol.	25
Inhibition of the Ras (oncogene) function.	29
Inhibition tyrosine kinases activity	30
Inhibition of cell transformation in mouse epidermal JB6 cell line.	31, 32
Anathematic effect	33
Analgesic effect	34
Hypotensive effect	25
Antioxidant activity Antiangiogenic effect in human placental veins	35, 36
Immunomodulation	37