Chemical Composition and Cytotoxic Activity of the Essential Oil and Oleoresins of In Vitro Micropropagated Ansellia africana Lindl: A Vulnerable Medicinal Orchid of Africa.

Orchids are rich treasure troves of various important phytomolecules. Among the various medicinal orchids, Ansellia africana stands out prominently in the preparing of various herbal medicines due to its high therapeutic importance. The nodal explants of A. africana were sampled from asymbiotically germinated seedlings on basal Murashige and Skoog (MS) medium and were micropropagated in MS medium supplemented with 3% sucrose and 10 µM meta topolin (mT) + 5 µM naphthalene acetic acid (NAA) +15 µM indole butyric acid (IBA) + 30 µM phloroglucinol (PG). In the present study, the essential oil was extracted by hydrodistillation and the oleoresins by the solvent extraction method from the micropropagated A. africana. The essential oil and the oleoresins were analysed by Gas Chromatography (GC) and GC/MS (Mass spectrometry). A total of 84 compounds were identified. The most predominant components among them were linoleic acid (18.42%), l-ascorbyl 2,6-dipalmitate (11.50%), linolenic acid (10.98%) and p-cresol (9.99%) in the essential oil; and eicosane (26.34%), n-butyl acetate (21.13%), heptadecane (16.48%) and 2-pentanone, 4-hydroxy-4-methyl (11.13%) were detected in the acetone extract; heptadecane (9.40%), heneicosane (9.45%), eicosane (6.40%), n-butyl acetate (14.34%) and styrene (22.20%) were identified and quantified in the ethyl acetate extract. The cytotoxic activity of essential oil and oleoresins of micropropagated A. africana was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide) assay on Vero cells compared to the standard drug doxorubicin chloride. The present research contains primary information about the therapeutic utility of the essential oil and oleoresins of A. africana with a promising future research potential of qualitative and quantitative improvement through synchronised use of biotechnological techniques.

Efficient Separation and Analysis of Triacylglycerols: Quantitation of ß-Palmitate (OPO) in Oils and Infant Formulas.

A high-efficiency, convenient, and reliable method for the separation of structurally similar triacylglycerols is detailed and applied in the quantitative analysis of 1,3-dioleoyl-2-palmitoylglycerol (OPO) in infant formulas and OPO oils. OPO is an important lipid component in "humanized" infant formula. A fast preparative isolation of an OPO-containing fraction from the crude complex mixture, by nonaqueous reversed phase HPLC, followed by Ag(+)-HPLC with detection at 205 nm allowed fine separation and detection of the desired fraction. OPO was quantitated independently of its regioisomer 1,2-dioleoyl-3-palmitoylglycerol (OOP) and isomers of stearoyl-linoleoyl-palmitoyl glycerol that might be present in infant formulas. For samples with low OPO content, an evaporative light-scattering detector (ELSD) was more preferable than UV detection, with a calculated LOD of 0.1 µg of OPO injected and LOQ of 0.3 µg. The method, which showed high reproducibility (RSD < 5%), was suitable for both high OPO content oils and low OPO products such as unenriched infant formula. A number of possible interference issues were considered and dealt with.

Androgenic effect of honeybee drone milk in castrated rats: roles of methyl palmitate and methyl oleate.

ETHNOPHARMACOLOGICAL RELEVANCE: Numerous honeybee (Apis mellifera) products have been used in traditional medicine to treat infertility and to increase vitality in both men and women. Drone milk (DM) is a relatively little-known honeybee product with a putative sexual hormone effect. The oestrogenic effect of a fraction of DM has recently been reported in rats. However, no information is available on the androgenic effects of DM. The purpose of the present study was to determine the androgen-like effect of DM in male rats and to identify effective compounds. MATERIALS AND METHODS: A modified Hershberger assay was used to investigate the androgenic effect of crude DM, and the plasma level of testosterone was measured. The prostatic mRNA and protein expression of Spot14-like androgen-inducible protein (SLAP) were also examined with real-time PCR and Western blot techniques. GC-MS and NMR spectroscopic investigations were performed to identify the active components gained by bioactivity-guided fractionation. RESULTS: The crude DM increased the relative weights of the androgen-dependent organs and the plasma testosterone level in castrated rats and these actions were flutamide-sensitive. DM increased the tissue mRNA and protein level of SLAP, providing further evidence of its androgen-like character. After bioactivity-guided fractionation, two fatty acid esters, methyl palmitate (MP) and methyl oleate (MO), were identified as active compounds. MP alone showed an androgenic effect, whereas MO increased the weight of androgen-sensitive tissues and the plasma testosterone level only in combination. CONCLUSION: The experimental data of DM and its active compounds (MO and MP) show androgenic activity confirming the traditional usage of DM. DM or MP or/and MO treatments may project a natural mode for the therapy of male infertility.

Cytotoxic steroids from the trunk of Berberis koreana.

A new steroid, itesmol 3-O-palmitate (1), along with two known steroids were isolated from the trunk of Berberis koreana. The structure of 1 was determined on the basis of spectroscopic analyses involving 2D NMR and chemical reactions. Compound 1 exhibited potential antiproliferative activity against A549, SK-OV-3, SK-MEL-2, and HCT-15 cell lines (respective IC(50) values of 7.41, 9.20, 4.53, and 12.91 µM).

Elimination of glycidyl palmitate in diolein by treatment with activated bleaching earth.

In this study, activated bleaching earth (ABE) was used to eliminate glycidyl esters from both triacyl- and diacylglycerol oils. To investigate the mechanism, glycerol dioleate containing glycidyl palmitate (GP) was treated with ABE and the fate of the GP was monitored by analyzing the feed, treated, and ABE-absorbed oils using a gas-liquid chromatograph equipped with a flame-ionized detector. GP was completely removed from both the treated and absorbed oils. This indicates that this treatment is useful for GE removal from diacylglycerol oil, although it was not achieved by absorption of GE on ABE but rather by modification of GP. The results of composition analysis demonstrate that GP is transformed to glycerol monopalmitate, glycerol palmitate oleate, and glycerol dipalmitate at a recovery rate of 99.1 ± 1.3 %. An increase in glycerol monooleate and trace amounts of free glycerol and fatty acids were also observed after treatment. The transformation is proposed to involve a ring-opening reaction of GP with water contained in the ABE and in the bulk oil followed by an interesterification reaction among the resultant monopalmitate and the glycerol dioleate of the bulk oil. All the generated compounds were simple acylglycerols and glycerol. Therefore, ABE treatment could be useful for GE removal during the manufacture of edible oils.

Chemical composition and antioxidant activity of the essential oil of endemic Viola tianshanica.

The composition and in vitro antioxidant activities of the essential oil and methanol extract of the aerial parts of Viola tianshanica were evaluated in this research. GC-MS analysis of the essential oil resulted in the identification of 15 constituents, representing 89.67% of the oil. The major compounds detected in the essential oil were dibutyl phthalate (15.19%), hexadecanoate methyl (8.65%), n-hexadecanoic acid (3.07%) and 2,3-pentanedione (2.62%). Essential oil and methanol extract were tested for their antioxidant activities using 1,1-diphenyl-2-picryl-hydrazyl free radical scavenging and ß-carotene linoleic acid assay. In addition, the total phenol of essential oil, polar subfraction and non-polar subfraction were determined.

Triterpenoids from Viburnum betulifolium.

Two new triterpenoids, ursa-12-sene-3ß,11ß-diol 3-O-palmitate (1) and ursa-12-sene-1ß,3ß,11α-triol 3-O-palmitate (2), were isolated from the 70% aqueous acetone extract of the aerial parts of Viburnum betulifolium, together with the artificial diene derivative of 2, ursa-12-dien-1ß,3ß-diol 3-O-palmitate (2a). Their structures were characterized by various spectroscopic methods, including 1D NMR, 2D NMR, and HR-ESI-MS.

Methyl palmitate, an acaricidal compound occurring in green walnut husks.

Walnut, Juglans regia L., is known for its insecticidal activities to a range of herbivores and microbes. Isolation and identification of bioactive compounds from walnut is a potential approach for the development of new pesticides. Laboratory experiments were carried out to investigate the acaricidal activities of green husk extracts of walnut. Bioassay-guided fractionation of petroleum-ether extracts of walnut led to the identification of a common plant-borne fatty acid ester, methyl palmitate (MP), which produced strong acaricidal activity (62.8% mortality) at 1 mg/ml at 24 h. The structure of MP was characterized with infrared spectrum and NMR, and the identification of MP confirmed using an authentic standard on high-performance liquid chromatography. Based on a slide dip bioassay, 10 mg/ml MP provided 97.9% mortality against adults of Tetranychus cinnabarinus (Boisduval) (Acari: Tetranychidae), whereas mortality against eggs was much lower (57.2%).

[Studies on chemical constituents from Grewia biloba].

OBJECTIVE: To investigate the chemical constituents of Grewia biloba and find its bioactive compounds. METHODS: Compounds were isolated by silica gel, MCI, Sephadex LH-20 and Rp-18 column chromatography, and purified by recrystallization. Their structures were elucidated by spectral analysis and other methods. RESULTS: Six compounds were isolated and identified as friedelin (1) epi-friedelan-3-ol (2), heneicosanoic acid (3), beta-sitosterol (4), Propyl palmitate (5), eatechin (6), respectively. CONCLUSION: Compounds 1-3, 5, 6 are isolated from Grewia genus for the first time and compounds 4 is isolated from Grewia biloba for the first time.

A new fatty acid ester of triterpenoid from Celastrus rosthornianus with anti-tumor activitives.

The methanol extract of the stalks of Celastrus rosthornianus Loes. afforded a new fatty acid ester of triterpene: 1beta,3beta-dihydroxy-olean-9(11),12-dienyl-3-palmitate (1), and three known compounds beta-amyrin palmitate (2), 3beta-hydroxy-11-oxo-olean-12-enyl-3-palmitate (3) and lupeol palmitate (4). Their structures were established by HRMS, 1D and 2D NMR experiments, as well as, by chemical degradation. The new compound showed cytotoxicity against human cervical squamous carcinoma (Hela) cells.

[Analysis of constituents of essential oil from the skin of water caltrop].

OBJECTIVE: To analyze the constituents of essential oil from the skin of water caltrop. METHOD: Water steam distillation and GC-MS were used. RESULT: 58 componds were separated respectively. 56 componds being identified which were 96. 5% of the totle essential oil. CONCLUSION: Diethyl phthalate, acetamide, N-acetyl-N, N'-1,2-ethanediylbis-, isopropyl palmitate, hexadecanoic acid, Z-11 and octadecanoic acid are the main component of essential oil from the skin of water caltrop.

Antioxidants and a new germacrane sesquiteroene from Carissa spinarum.

A new germacrane derivative, care none (1) was isolated from the stems of a well-known medicinal plant, Carissa spinarum L. together with a new ester, 3'-(4''-methoxyphenyl)-3-oxo-ropionyl hexadecanoate (2). Their structures were established using 1D, 2D NMR spectroscopic and synthetic methods. The chloroform extract of the plant displayed strong antioxidant (DPPH) activity. Several other known compounds were also isolated for the first time from this active extract. The antioxidant activity of major lignans was studied.

A potent anti-complementary acylated sterol glucoside from Orostachys japonicus.

In order to isolate substances that inhibit the hemolytic activity of human serum against erythrocytes, we have evaluated whole plants of the Orostachys japonicus species with regard to its anti-complement activity, and have identified its active principles following activity-guided isolation. A methanol extract of the O. japonicus, as well as its n-hexane soluble fraction, exhibited significant anti-complement activity on the complement system, which was expressed as total hemolytic activity. A bioassay-guided chromatographic separation of the constituents resulted in the isolation of three known compounds 1-3 from the active n-hexane fraction. The structure of these compounds were analyzed, and they were identified as hydroxyhopanone (1), beta-sitosteryl-3-O-beta-D-glucopyranosyl-6'-O-palmitate (2), and beta-sitosteryl-3-O-beta-D-glucopyranoside (3), respectively. Of these compounds, compound 2 exhibited potent anti-complement activity (IC50= 1.0 +/- 0.1 microM) on the classical pathway of the complement, as compared to tiliroside (IC50= 76.5 +/- 1.1 microM), which was used as a positive control. However, compounds 1 and 3 exhibited no activity in this system.

Two new triterpene esters from the twigs of Brachylaena ramiflora from the Madagascar rainforest.

Bioassay-guided fractionation of a CH(2)Cl(2)/MeOH extract of the small twigs of Brachylaena ramiflora var. ramiflora resulted in the isolation of the two new triterpene esters 1 and 2 and five known triterpenoids, alpha-amyrin palmitate (3), beta-amyrin palmitate (4), beta-amyrin acetate (5), lupeyl acetate (6), and lupeol (7). The structures of the two new compounds were established as kairatenyl palmitate (1) and hopenyl palmitate (2) on the basis of 1D and 2D NMR spectroscopic data interpretation and chemical conversions. All the isolated compounds showed weak cytotoxicity against the A2780 human ovarian cancer cell line.

[Chemical constituents of Aster albscens Hand.-Mazz].

Four compounds, stigmasterol, D-hurulon-14-en-3-ol, stigma-delta7.22-dien-3beta-palmitate and friedelin were isolated from the root of Aster albscens for the first time. Their structures were elucidated by spectral methods.

3 beta-O-palmityl longispinogenin from Trichocereus chilensis.

From the aerial parts of Trichocereus chilensis a new triterpenetriol fatty acid ester was isolated. Its structure was shown to be 3 beta-O-palmityl longispinogenin (olean-12-ene-3 beta,16 beta,28-triol-3-palmitate) [1] by chemical and spectroscopic methods. Compound 1 is inactive in brine shrimp lethality and cytotoxicity bioassays.