The effects ofEurycoma longifolia Jack were studied on the sexual behaviour of male rats. Sexually normal male rats were treated twice daily with 500 mg kg(-1) of different fractions ofE. longifolia Jack for 10 days prior to test and were then observed for their copulatory behaviour with a receptive female in a copulation cage. Results showed that was a significant increase (p<0.05) in EL-1, EL-2, EL-3 but significant decrease (p<0.05) in both PEI-1 and PEI-2 in treated male rats as compared to the control male rats indicating thatE. longifolia Jack increased the sexual performance of the treated male rats by extending the duration of coitus and decreasing the refractory period between the different series of copulation. Hence, this preliminary work supports the folk use of this plant as having aphrodisiac property.
An analytical method using HPLC with UV detection was developed to investigate the quassinoid content of Eurycoma longifolia Jack (Simaroubaceae) collected from various sources. Eurycomanone (1), longilactone (2), 14,15beta-dihydroxyklaineanone (3), 15beta-acetyl-14-hydroxyklaineanone (4), 6alpha-hydroxyeurycomalactone (5), and eurycomalactone (7) were isolated as reference standards and together with the synthesized 1beta,12alpha,15beta-triacetyleurycomanone (6, internal standard), were identified by NMR, MS, UV and IR spectroscopies. Their coefficient of variation values for 0.50-35 microg ml(-1) concentrations of quassinoids and their retention times measured within- and between-day were small. The recoveries of the spiked quassinoids in E. longifolia samples and their detection limits at 8.5 times signal to noise ratio were 99.75-109.13% and 0.01 microg ml(-1), respectively. From the root samples analysed, 1 had the highest concentration, being about 16.8-39.6 fold higher than the other quassinoids 2, 3, 5, 7 but 145.3 fold higher than 4 which showed the lowest concentration.
The effects of various fractions of Eurycoma longifolia Jack were studied on the orientation activities of the inbred, adult middle-aged Sprague-Dawley rats, 9 months old and retired breeders towards the receptive females (anogenital sniffing, licking, mounting), the environment (climbing, raring, exploration), themselves (nongenital grooming, genital grooming) and mobility (restricted, unrestricted) after treating these subjects twice daily for 10 days. Results showed that subjects treated with 800 mg/kg of E. longifolia Jack increased orientation activities towards the receptive females (anogenital sniffing, licking and mounting), increased genital grooming towards themselves and restricted movements to a particular area of the cage but decreased interest in the external environment (climbing, raring, exploration) as compared with the controls during the investigation period. In conclusion, this study gives further evidences that different fractions of E. longifolia Jack modified the orientation activities of the middle-aged male rats.
The effect of increasing doses of various fractions of Eurycoma longifolia Jack extracts on libido was examined in middle-aged male rats. The results showed that a high dose (800 mg/kg) of all E. longifolia Jack extracts significantly increased mount frequency (MF) (P < 0.05) over that of untreated controls, but had no effect on the frequency of intromission or ejaculation. Methanol, chloroform, water, and butanol fractions exhibited MF of 2.5 +/- 0.1, 2.6 +/- 0.3, 2.5 +/- 0.1 and 2.6 +/- 0.2, respectively, in adult, middle-aged male rats, and retired breeders versus 2.3 +/- 0.1 in untreated controls. This translated to a minor increase in MF of 8.7%, 13.0%, 8.7%, and 13.0% for these fractions, respectively, during the 20-minute observation period. The results of this study show that E. longifolia Jack extracts can increase libido in middle-aged male rats.
Eurycoma longifolia Jack. is a treelet that grows in the forests of Southeast Asia and is widely used throughout the region because of its reported medicinal properties. Widespread harvesting of wild-grown trees has led to rapid thinning of natural populations, causing a potential decrease in genetic diversity among E. longifolia. Suitable genetic markers would be very useful for propagation and breeding programs to support conservation of this species, although no such markers currently exist. To meet this need, we have applied a genome complexity reduction strategy to identify a series of single nucleotide polymorphisms (SNPs) within the genomes of several E. longifolia accessions. We have found that the occurrence of these SNPs reflects the geographic origins of individual plants and can distinguish different natural populations. This work demonstrates the rapid development of molecular genetic markers in species for which little or no genomic sequence information is available. The SNP markers that we have developed in this study will also be useful for identifying genetic fingerprints that correlate with other properties of E. longifolia, such as high regenerability or the appearance of bioactive metabolites.
The effects of Eurycoma longifolia Jack were studied on the sexual qualities of middle aged male rats after dosing them with 0.5 g/kg of various fractions of E. longifolia whilst the control group received 3 ml/kg of normal saline daily for 12 weeks. Results showed than E. longifolia Jack enhanced the sexual qualities of the middle aged male rats by decreasing their hesitation time as compared to controls with various fractions of E. longifolia Jack produced 865-916 (91-96), 860-914 (92-98), 850-904 (93-99), 854-890 (95-99), 844-880 (94-98), 840-875 (94-98), 830-870 (94-98), 825-860 (94-98), 820-850 (96-99), 800-840 (93-98), 750-795 (94-99) and 650-754 sec (82-95%) in contrast to controls which produced 950 (100), 934 (100), 910 (100), 900 (100), 895 (100), 890 (100), 885 (100), 880 (100), 855 (100), 860 (100), 800 (100) and 790 sec (100%) throughout the investigation period. Besides these, there was a transient increase in the % of the male rats responding to the right choice after chronic administration of 0.5 g/kg E. longifolia Jack, with more than 50% of the male rats scored right choice after 2 weeks post-treatment and the effect was more prominent at the dose of the observation period. However, there was no sexual enhancement of the middle aged male rats which consumed normal saline since only 45-55% of the male rats responded to right choice throughout the investigation period. Hence, this study shows that E. longifolia Jack enhanced the sexual qualities of the middle aged male rats, further supports the folkuse of E. longifolia Jack as an aphrodisiac.
Eurycoma longifolia Jack was investigated for sexual motivation activity in adult, middle-aged male mice and in retired breeders, using the modified open field and the modified runway choice methods. Each mouse received 500 mg/kg of one of 4 fractions of E. longifolia Jack, viz. chloroform, methanol, butanol, and water, whereas the mice in the control and yohimbine groups received 3 ml/kg of normal saline and 30 mg/kg of yohimbine daily respectively for 10 d. The results show a transient increase in the percentage of male mice responding to the right choice after chronic consumption of the fractions with 50 percent of the adult middle-aged male mice treated with E. longifolia Jack and yohimbine scoring the right choice after 8 and 5 days post-treatment respectively. In conclusion, this study has shown that E. longifolia Jack continues to enhance sexual motivation in adult, middle-aged male mice and in retired breeders.
The roots of Eurycoma longifolia Jack have been used as traditional medicine to treat malaria. A systematic bioactivity-guided fractionation of this plant was conducted involving the determination of the effect of its various extracts and their chemical constituents on the lactate dehydrogenase activity of in vitro chloroquine-resistant Gombak A isolate and chloroquine-sensitive D10 strain of Plasmodium falciparum parasites. Their antiplasmodial activity was also compared with their known in vitro cytotoxicity against KB cells. Four quassinoids, eurycomanone (1), 13,21-dihydroeurycomanone (3), 13 alpha(21)-epoxyeurycomanone (4), eurycomalactone (6) and an alkaloid, 9-methoxycanthin-6-one (7), displayed higher antiplasmodial activity against Gombak A isolate but were less active against the D10 strain when compared with chloroquine. Amongst the compounds tested, 1 and 3 showed higher selectivity indices obtained for the cytotoxicity to antiplasmodial activity ratio than 14,15 beta-dihydroxyklaineanone (2), eurycomanol (5), 6 and 7.
Eurycoma longifolia Jack commonly known as Tongkat Ali in Malaysia, has been used in Malaysia to increase male virility and sexual prowess. The objective of this study is to evaluate sexual arousal in sexually sluggish old male rats, 24 months old and retired breeders, receiving 200, 400, or 800 mg/kg of various fractions of E. longifolia Jack, twice daily, for 10 days. Control rats received 3 ml/kg of normal saline. The aphrodisiac effect was monitored by the act of yawning and stretching because yawning, either alone or associated with stretching, is considered an ancestral vestige surviving throughout evolution that promotes sexual arousal. The results showed that 800 mg/kg of E. longifolia Jack increased yawning by 50% and stretching by 16.7% in sexually sluggish old male rats, by 676-719% and 31-336%, respectively, in sexually active male rats, and by 22-44% and 75-100%, respectively, in middle aged, 9 months old and retired breeders. We conclude that the results of this study support the folk use of this plant as an aphrodisiac.
The DCA (Drug Control Authority), Malaysia, has implemented the phase 3 registration of traditional medicines on 1 January 1992, with special emphasis on the quality, efficacy, and safety (including the presence of heavy metals) in all pharmaceutical dosage forms of traditional medicine preparations. As such, a total of 100 products in various pharmaceutical dosage forms of a herbal preparation, containing Tongkat Ali, were analyzed for mercury content using cold vapor atomic absorption spectrophotometer. Results showed that 36% of the above products possessed 0.52 to 5.30 ppm of mercury and, therefore, do not comply with the quality requirement for traditional medicines in Malaysia. Out of these 36 products, 5 products that possessed 1.05 to 4.41 ppm of mercury were in fact have already registered with the DCA, Malaysia. However, the rest of the products that contain 0.52 to 5.30 ppm of mercury still have not registered with the DCA, Malaysia. Although this study showed that only 64% of the products complied with the quality requirement for traditional medicines in Malaysia pertaining to mercury, they cannot be assumed safe from mercury contamination because of batch-to-batch inconsistency.
A validated HPLC analysis of eurycomanone (1), a bioactive quassinoid, in rat plasma following oral and intravenous administration of Eurycoma longifolia Jack extract was developed for pharmacokinetic and bioavailability studies. Relatively high plasma eurycomanone concentrations were detected after an intravenous injection of 10 mg/kg extract F2 containing 1.96 mg/kg of the quassinoid. However, it declined rapidly to zero after 8 h. Its mean elimination rate constant (k(e)), biological half-life (t(1/2)), volume of distribution (V(d)) and clearance (CL) were 0.88 +/- 0.19 h (-1), 1.00 +/- 0.26 h, 0.68 +/- 0.30 L/kg and 0.39 +/- 0.08 L/h/kg, respectively. Following oral administration of eurycomanone, its Cmax and Tmax values were detected as 0.33 +/- 0.03 microg/mL and 4.40 +/- 0.98 h, respectively. The plasma concentration of the quassinoid after oral administration was much lower than after intravenous application in spite of the oral dose being 5 times higher. The results indicate that eurycomanone is poorly bioavailable when given orally. A comparison of the AUC (0-->infinity) obtained orally to that obtained after an intravenous administration (normalized for dose differences) revealed that the absolute bioavailability of the compound was low with 10.5 %. Furthermore, the compound appeared to be well distributed in the extravascular fluids because of its relatively high V(d) value. The poor oral bioavailability was not attributed to instability problems because eurycomanone has been shown to be stable under different pH conditions. Thus, its poor oral bioavailability may be due to poor membrane permeability in view of its low P value and/or high first-pass metabolism.
Among the quassinoids isolated from Eurycoma longifolia Jack, eurycomanone was identified as the most potent and toxic inhibitor of the chloroquine-resistant Gombak A isolate of Plasmodium falciparum. Several diacylated derivatives of eurycomanone, 1,15-di-O-isovaleryleurycomanone, 1,15-di-O-(3,3-dimethylacryloyl)- eurycomanone and 1,15-di-O-benzoyleurycomanone were synthesized by direct acylation with the respective acid chlorides. The monoacylated 15-O-isovaleryleurycomanone was synthesized by selective protection of the other hydroxy groups of eurycomanone with trimethylsilyl trifluoromethanesulphonate to enable the exclusive acylation of its C-15 hydroxy group. This was followed by the removal of the protecting groups with citric acid. The diacylated eurycomanones exhibited lower antiplasmodial activity against the Gombak A isolates and lower toxicity in the brine shrimp assay when compared to eurycomanone. In contrast, the monoacylated derivative displayed comparable antiplasmodial potency to eurycomanone, but its toxicity was reduced. Thus, preliminary studies of the synthesized acylated eurycomanones have shown that acylation only at the C-15 hydroxy group may be worthy of further antimalarial investigation.
The methanol, n-butanol, chloroform and water extracts obtained from the root of Eurycoma longifolia Jack were assayed using methylene blue assay to evaluate its cytotoxic effect against KB, DU-145, RD, MCF-7, CaOV-3, MDBK cell lines. The results showed that all the root extracts except the water extract of E. longifolia produced significant cytotoxic effect on these cell lines. However, no significant cytotoxic effect was detected on MDBK (kidney) normal cell line. 9-methoxycanthin-6-one, an alkaloid, was detected in each extract with different intensities by reversed-phase high performance liquid chromatography.
In the present study we examined the effect of E. longifolia methanol extract (TA164) on the GSH levels of P. falciparum infected erythrocytes and uninfected erythrocytes. Our study on parasite growth shows the IC50 and IC75 values of TA164 to be 0.17 g/ml and 6 g/ml respectively while for BSO was 25.5 g/ml and 46.5 g/ml respectively. About 95% to 100% growth inhibition of P. falciparum infected erythrocyte was observed when treated with TA164 and BSO at 16 g/ml and 64 g/ml respectively. The study on GSH contents indicated that non-infected erythrocytes treated with 6 g/ml (IC75 values) of TA164 at 24 hours incubation showed less GSH content as compared to non-treated erythrocytes. A similar observation was seen on treated trophozoite infected erythrocyte (10% parasitemia) when treated with 6 g/ml at 3 hours incubation. Analysis of the GSH contents of parasite compartments treated with TA164 at the same concentration (6 g/ml) for 3 hours incubation indicated a reduction of GSH contents. At the same concentration, TA164 did not affect the GSH contents of enriched trophozoite infected erythrocytes (60-70% parasitemia). TA164 did affect the GSH content of non-infected erythrocyte at 24 hours (accept IC50 value) as well as the parasite compartments (trophozoite infected erythrocyte and parasite itself) but fails to affect the GSH content of enriched trophozoite infected erythrocyte.
Extracts of the plant Eurycoma longifolia have been shown to possess cytotoxic, antimalarial, anti-ulcer, antipyretic and plant growth inhibition activities. The present study investigated the effects of extracts and their chromatographic fractions from the root of E. longifolia on the growth of a human breast cancer cell line, MCF-7. Our data indicated that E. longifolia extracts and fractions exert a direct antiproliferative activity on MCF-7. The bioassay-guided root fractionation resulted in the isolation of three active fractions, F5, F6 and F7, which displayed IC50 values of (6.17+/-0.38) microg/ml, (4.40+/-0.42) microg/ml and (20.00+/-0.08) microg/ml, respectively. The resultant from F7 purification, F16, exhibited a higher cytotoxic activity towards MCF-7, (IC50=15.23+/-0.66 microg/ml) and a certain degree of selectivity against a normal breast cell line, MCF-10A (IC50=66.31-0.47 microg/ml). F16 significantly increased apoptosis in MCF-7 cells, as evaluated by the Tdt-mediated dUTP nick end labelling assay and nuclear morphology. Western blotting revealed down-regulation of the anti-apoptotic Bcl-2 protein expression. F16, however, did not affect the expression of the pro-apoptotic protein, Bax. These results, therefore, suggest that F16 has antiproliferative effects on MCF-7 cells by inducing apoptosis through the modulation of Bcl-2 protein levels.
The DCA (Drug Control Authority), Malaysia has implemented the phase three registration of traditional medicines on 1 January 1992. As such, a total of 100 products in various pharmaceutical dosage forms of a herbal preparation found in Malaysia, containing tongkat Ali hitam, either single or combined preparations, were analyzed for the presence of a heavy toxic metal, mercury, using atomic absorption spectrophotometer, after performing a simple random sampling to enable each sample an equal chance of being selected in an unbiased manner. Results showed that 26% of these products possessed 0.53-2.35 ppm of mercury, and therefore, do not comply with the quality requirement for traditional medicines in Malaysia. The quality requirement for traditional medicines in Malaysia is not exceeding 0.5 ppm for mercury. Out of these 26 products, four products have already registered with the DCA, Malaysia whilst the rest, however, have not registered with the DCA, Malaysia.
Eurycoma longifolia, locally known as 'Tongkat Ali' is a popular local medicinal plant that possess a lot of medicinal properties as claimed traditionally, especially in the treatment of malaria. The claims have been proven scientifically on isolated compounds from the plant. The present study is to investigate the anti malaria properties of Eurycoma longifolia standardized extract (root) (TA164) alone and in combination with artemisinin in vivo. Combination treatment of the standardized extract (TA164) with artemisinin suppressed P. yoelii infection in the experimental mice. The 4 day suppressive test showed that TA164 suppressed the parasitemia of P. yoelii-infected mice as dose dependent manner (10, 30 and 60 mg/kg BW) by oral and subcutaneous treatment. By oral administration, combination of TA164 at 10, 30 and 60 mg/kg BW each with artemisinin respectively showed a significant increase in the parasitemia suppression to 63, 67 and 80 percent as compared to artemisinin single treatment (31%). Using subcutaneous administration, at 10 mg/kg BW of TA164 in combination with 1.7 mg/kg BW of artemisinin gave a suppression of 80% of infection. This study showed that combination treatment of TA164 with artemisinin gives a promising potential anti malaria candidate using both oral and subcutaneous route, the later being the most potent.
Objective: To evaluate methanolic, ethanolic, acetone and aqueous extracts from different parts of Eurycoma longifolia (E. longifolia) (leave, stem, and root) for antibacterial activity against Gram-positive and Gram-negative bacteria and to utilize the leaves and stem parts rather than the root, which is already used for male sexual enhancement in Malaysia.
Methods: The study took place in the Laboratory of Molecular Biology of Biotechnology Engineering Department, Malaysia between January 2005 and June 2006. Methanolic, ethanolic, acetone and aqueous extracts of leaves, stems and roots of E. longifolia were investigated for their antibacterial properties using Agar-well diffusion method.
Results: The alcoholic and acetone extracts of the leaves and stem extracts were active on both Gram-positive and Gram-negative bacteria except against 2 strains of Gram-negative bacteria (Escherichia coli and Salmonella typhi). The root extracts had no antibacterial activity against Gram-positive and Gram-negative bacteria tested. Aqueous leaves extract showed antibacterial activity against Staphylococcus aureus and Serratia marscesens.
Conclusion: The alcoholic and acetone extracts from leaves and stems of E. longifolia contain potent antibacterial agent(s). This plant can serve as a potential source of antibacterial compounds.
Three popular medicinal plants regarded as improving human sexual function in some parts of Southeast Asia were analysed for their mutagenic properties using modified Ames test (fluctuation test). Extract of one of the plants, Tacca integrifolia Ker-Gawl., was found to be mutagenic using Salmonella typhimurium strains TA98 and TA100. Extract of T. integrifolia, Eurycoma longifolia Jack and Helmintostachys zeylanica (L.) Hook were cytotoxic to human cell lines, Hep2 and HFL1, with IC50 ranging from 11 mug/ml to 55 mug/ml. Extract of E. longifolia was the most cytotoxic with IC50 of 11 mug/ml and 13 mug/ml on Hep2 and HFL1 cell lines respectively. Combined extract of T. integrifolia and H. zeylanica was more cytotoxic than single extract on both Hep2 and HFL1 cell lines while combined extract of E. longifolia and H. zeylanica was more cytotoxic than single extract on Hep2 cell lines. Under the conditions of this study it can be concluded that T. integrifolia is mutagenic and the combined extracts of the medicinal plants was highly cytotoxic.