ORIGINAL ARTICLE Nutraceutical properties of Thai "Yor", Morinda citrifolia and "Noni" juice extract Pranee Nandhasri1, Kammal Kumar Pawa2, Jamnong Kaewtubtim2, Chareonchai Jeamchanya3, Chalerm Jansom4 and Chisanucha Sattaponpun4 Abstract Nandhasri, P., Kumar Pawa K., Kaewtubtim J., Jeamchanya C., Jansom C. and Sattaponpun C. Nutraceutical properties of Thai "Yor", Morinda citrifolia and "Noni" juice extract Songklanakarin J. Sci. Technol., 2005, 27(Suppl. 2) : 579-586 Morinda citrifolia Linn. is known in Thailand as "Yor" and commonly called "Noni" in Hawaii, Tahiti and USA. This paper reports the results of chemical analysis of Thai "Yor" (M. citrifolia) juice extract. The juice from "Yor" fruit, was found to be high in vitamin C, vitamin B-complex contents (HPLC method), and was also high in anti-oxidants, anthraquinones, flavonoids, saponins and scopoletin (TLC method). Sodium, potassium, calcium, iron and selenium contents of the juice (AAS method) are also reported. All the results of Thai "yor" juice are comparable to those of the commercially distributed 'Noni' juice. The Thai "Yor" juice was free of microorganisms and could safely be consumed in the recommended amount of 30 ml/day. The results of this study only apply to the Thai "Yor" juice preparation. The commercial "Noni" juice may be processed differently and may contain other ingredients. Key words : Nutraceutical and functional food, Morinda citrifolia, Noni juice extract, HPLC, TLC 1M.A.(Chemistry), Asst. Prof., 2M.D., Asst. Prof., 3M.D., 4B.Sc.(Chemistry), Research Center, Faculty of Medicine, Thammasat University, Rangsit Campus, Pathum Thani, 12120 Thailand. Corresponding e-mail: [email protected] Received, 6 August 2004 Accepted, 24 November 2004 Songklanakarin J. Sci. Technol. 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À√◊Õ "¬Õ" ¡’™◊ËÕ‡√’¬°«à“ "‚ππ‘" „πÀ¡Ÿà‡°“–Œ“«“¬ ·≈–µ“Œ‘µ‘ °“√«‘®—¬π’È√“¬ß“π º≈°“√«‘‡§√“–Àå∑“߇§¡’„ππÈ” °—¥º≈¬Õ¥â«¬«‘∏’ HPLC æ∫«à“¡’«‘µ“¡‘π´’·≈–«‘µ“¡‘π∫’√«¡„πª√‘¡“≥ Ÿß ¡’ “√°≈ÿà¡ µâ“πÕπÿ¡Ÿ≈Õ‘ √– ·Õπ∑√“§«‘‚ππ ø≈“‚«πÕ¬ ´“‚ªπ‘π ·≈– ‚§‚æ‡≈∑‘π ´÷Ëß«‘‡§√“–Àå¥â«¬«‘∏’ TLC ·≈–æ∫«à“¡’ ‚´‡¥’¬¡ ‚ª·µ ‡´’¬¡ ·§≈‡´’¬¡ ‡À≈Á° ·≈–‡´‡≈‡π’¬¡ ‡¡◊ËÕ«‘‡§√“–Àå¥â«¬«‘∏’ AAS º≈°“√«‘‡§√“–ÀåπÈ” °—¥¬Õ‰∑¬ ‡∑’¬∫‰¥â°—∫πÈ” °—¥∑’˺≈‘µ‡ªìπ°“√§â“ "‚ππ‘" πÈ”¬Õ¢Õ߉∑¬π’Ȫ√“»®“° “√®ÿ≈‘π∑√’¬å∑’ˇªìπæ‘… ·≈–ª≈Õ¥¿—¬µàÕ°“√ ∫√‘‚¿§ ´÷Ëß·π–π”„Àâ¥◊Ë¡«—π≈– 30 ¡≈. º≈°“√»÷°…“π’ȇªìπº≈°“√√“¬ß“π‡©æ“–πÈ”¬Õ∑’ˇµ√’¬¡¢÷Èπ‡Õßπ’ȇ∑à“π—Èπ πÈ”¬Õ ∑’˺≈‘µ‡ªìπº≈‘µ¿—≥±å∑“ß°“√§â“Õ“®¡’°√–∫«π°“√º≈‘µ∑’Ë·µ°µà“ßÕÕ°‰ª ·≈–Õ“®¡’°“√‡µ‘¡ à«πº ¡Õ◊Ëπ¥â«¬ Àπ૬«‘®—¬ ¡ÿπ‰æ√·≈–Õ“À“√ ‚§√ß°“√®—¥µ—Èß»Ÿπ¬å«‘®—¬ §≥–·æ∑¬»“ µ√å ¡À“«‘∑¬“≈—¬∏√√¡»“ µ√å Õ”‡¿Õ§≈ÕßÀ≈«ß ®—ßÀ«—¥ ª∑ÿ¡∏“π’ 12120 Nutraceutical and functional food (NFF) that has helped relieved people (n ≥ 10,000) from products are increasingly becoming health products the suffering of about 22 conditions, such as of choice for the next decade. Nutraceutical (or arthritis, heart disease, diabetes, headache and part of the food) offers medical or health benefits muscle pain, high blood pressure, cancer, etc. to the consumer by providing a means for the (Solomon, 1999). maintenance of health and well being and protect- Recently 'Noni' juice extract has been ion from diseases. A functional food provides the commercially processed and distributed inter- body with the required amounts of vitamins, fats, nationally as a dietary supplement. Relatively protein, carbohydrates and many other compounds unheard of 5 years ago, M. citrifolia has now that are needed for its survival (DeFelice, 2002; exploded into nearly a billion-dollar industry. Kalra, 2003). The objectives of this paper are (1) to Morinda citrifolia Linn. an Indian mulberry determine the nutraceutical components of Thai or "Noni" in English, and"Yor" in Thai, is the plant "Yor" (M. citrifolia) juice extract, which was that can be used as a raw material for nutraceutical prepared using a Thai traditional method, and (2) and functional food products. In Thailand, "Yor" to compare the nutraceutical composition of Thai leaves cooked with curry give a wonderful and Yor Noni juice extract with the U.S. commercial nutritious dish. "Yor" fruits can be prepared as a product. Thai traditional salad "Somtumyor". Dried fruits or leaves powder boiled with water produces a tea, Materials and Methods which is used to relief blood pressure, muscle pains and vomiting. Yor fruits were obtained from several There are more than 120 nutraceutical provinces in Thailand, i.e. Phra Nakhon Si compounds identified in Noni (Solomon, 1999). Akutthaya, Nakhon Pathom, Nonthaburi, Pathum Noni or Yor juice extract which is obtained from Thani, Nakhon Ratchasima, Phrae, Saraburi and fermented Noni fruits is the most effective product Surat Thani. The fresh fruits were cleaned by Songklanakarin J. Sci. Technol. Nutraceutical properties of Thai "Yor" Vol. 27 (Suppl. 2), 2005 : Thai Herbs 581 Nandhasri, P., et al. washing with water. The bioactive Yor juice (BYJ) method. (Wagner, 1993 and Wagner, 2000). The extract was prepared by mixing the clean and mobile phase system, spraying reagents, UV and mature fruits with red sugar at the proportion of visible detection reults are summarized in Table 1. 3:1 in a glass or stainless steel reactor. The mixture The microplates for TLC determination were made was often stirred during the first period of 2 weeks by dipping a sandwich of micro slide in a mixture then left in the reactor until the foul smell of Yor of Silica gel 60 G and water at the ratio of 1:1. dis-appeared, which took about 3-6 months. The Sodium, potassium, calcium, iron and BYJ extract was separated from the residue by selenium contents were analyzed using Atomic filtering through cotton cloth. Absorption Spectrophotometric (AAS) method The BYJ extracts were analyzed with High (AOAC, 1995). The quantities of fat, protein, Performance Liquid Chromatography, (HPLC), to carbohydrate and calories were analyzed using obtain the contents of vitamins C and B-complex. AOAC methods (AOAC, 1995; Pendl, 1998). The HPLC system was composed of Spherisorb S5 microorganisms were analyzed by Bam's method ODS2, 25 cm Cartridge, 4.6 mm diameter column (U.S. FDA., 1998). and UV-detector at 254 nm with the isocratic mobile phase of 0.05 M sodium acetate, (pH 5.2) Results, Discussion and Conclusion acetonitrile and 0.005 M hexanesulfonic acid sodium salt monohydrate (HSA) at the ratio of The results of vitamins analysis in of bio- 60:28:12 (Nandhasri,1986). active Yor juice (BYJ) extract are comparable to The characteristic components of BYJ those of commercial Noni juice (NJ). The vitamin extract i.e. anti-oxidants, alkaloids, anthraquinones, content in bio-active Yor juice (BYJ) extract was coumarins, essential oils, flavonoids, saponins, in the range of 38.54-97.64 mg/100 g. It is scopoletin and sugars were obtained by TLC interesting to note that the vitamin content of BYJ Figure 1. The comparative results of vitamins content: vitamin C, vitamin B1, vitamin B2, vitamin B3 and vitamin B12 in Morinda citrifolia L. of Bio-active Yor Juice (BYJ) extract and Noni juice product from abroad (NJ). Vol. 27(Suppl.2),2005:ThaiHerbs Songklanakarin J.Sci.Technol. Table 1. Thin Layer Chromatography Methods (Wagner, 1993 and Wagner, 2000). Summarized mobile phase system, spraying reagent, and colour result of UV and visible detection. No. Test Extraction Mobile phase Spraying reagent Detection Result 1 Alkaloids 10 ml methanol+1ml 10% Toluene/Ethyl acetate/ Dragendroff reagent Visible orange ammonia Diethylamine (70:20:10) 2 Anthraqui- 10 ml methanol Ethyl acetate/Methanol/Water KOH reagent Visible and orange/red nones (100:13.5:10) UV 366 nm 3 Antioxidants 10 ml dichloromethane Ethyl acetate/Toluene (50:50) DPPH reagent Visible white spot on purple background 4 Coumarins 10 ml dichloromethane Toluene / Ethyl acetate 1. None Visible and light blue/ (93: 7) UV 366 nm purple/green 2. KOH reagent UV 366 nm Light green 5 Essential oils 10 ml dichloromethane Toluene / Ethyl acetate Anisaldehyde - Heat 1000C pink/purple/red 93: 7) Sulfuric acid reagent Visible /orange/green 582 /black 6 Flavonoids 10 ml methanol Ethyl acetate/Formic Natural products- UV 366 nm orange/yellow acid/Acetic acid/Water Poly ethyleneglycol /green/blue (100:11:11:27) reagent 7 Glycosides 10 ml 50%methanol Ethyl acetate/Methanol/ Kedde reagent Visible pink/purple +10%lead acetate and Water Thai "Yor" of Nutraceutical properties extract by chloroform (81:11:8) 8 Phenolic 10 ml dichloromethane Toluene / Ethyl acetate Ferric chloride Visible Blue-black/ compounds (93: 7) reagent Green-black 9 Saponins 10 ml methanol and Chloroform/Methanol/ Vanillin-Sulfuric Visible blue extract by Water acid reagent 1 ml water + 3ml butanol (64:50:10) 10 Scopoletin 5 ml methanol+ 5 ml Ethyl acetate/Methanol/ - UV 366 nm light blue Nandhasri, P., dichloromethane Water (100:6:4) 11 Sugars 10 ml water Acetonitrile/Water 10 % Sulfuric acid Heat 1000C gray (4:1) Visible et al. Songklanakarin J. Sci. Technol. Nutraceutical properties of Thai "Yor" Vol. 27 (Suppl. 2), 2005 : Thai Herbs 583 Nandhasri, P., et al. Figure 2. The TLC chromatograms show the characteristic of organic substances of the BYJ extract products: anti-oxidant, saponins, scopoletin and sugar. Figure 3. The TLC chromatograms show the characteristic of organic substances of the BYJ extract products: alkaloids, anthraquinones, essential oils and Flavonoids. of Thai Yor was higher than that of Noni juice ure 2). Essentials oils were found to be moder- (NJ) from the U.S. (Figure 1). ately high, while anthraquinones, alkaloids and The characteristics of the organic sub- flavonoids were low (Figure 3).