17th Philippine Chemistry Congress
May 23-25, 2001
Cagayan de Oro City

ABSTRACTS: NATURAL PRODUCTS CHEMISTRY

TS1 NP1

Antimicrobial Compounds from Mentha cordifolia

Consolacion Y. Ragasa1, Rose Dumag1 and John A. Rideout2

1Chemistry Department, De La Salle University, Taft Avenue, Manila

2Chemistry Department, Central Queensland University, Rockhampton, Queensland, Australia

The dichloromethane extract of Mentha cordifolia afforded 5, 6, 4’-trihydroxy-7,8,3’-trimethoxyflavone (1), piperitenone epoxide (2), lutein and sitosterol. The structures of 1 and 2 were elucidated by extensive 1D and 2D NMR techniques. Antimicrobial tests were conducted on 1 and 2 by the agar cup method. Compound 1 was found active against B. subtilis, P. aeruginosa, C. albicans, T. mentagrophytes and A. niger, but inactive against E. coli and S. aureus. Compound 2 was active against P. aeruginosa, C. albicans, T. mentagrophytes and A. niger, but inactive against E. coli, S. aureus and B. subtilis.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TS1 NP2

COMPARISON OF ILANG-ILANG (TIAONG ACCESSION) OIL EXTRACTED BY SUPERCRITICAL FLUID EXTRACTION AND HYDRODISTILLATION

Hidelisa P. Henrnandez and Tzarina Marie F. Trio

Institute of Chemistry, University of the Philippines Los Baņos, Laguna

Essential oil from flowers of ilang-ilang (Cananga odorata {Lamark} Hook Fil et. Thomson) was extracted by supercritical fluid extraction (SFE) using carbon dioxide as solvent. The SFE oil was compared with the hydrodistilled oil in terms of yield, physical properties, gas and sensory analysis. The source of all the oils analyzed were accession plants maintained by the Plantation Crops Division, Dept. of Horticulture, UPLB.

The best SFE oil was obtained by extracting at 40oC and 1700 psi. The yield (wt/wt) of SFE and hydrodistillation were 0.43% and 0.90%, respectively. Both extracts were clear yellowish but the SFE oil was more viscous and had a finer floral scent.

Gas chromatography-mass spectrometry revealed that both extracts had 13 constituents but differed in terms of the relative proportion of the constituents. Tentative identification of the constituents and calculation of peak- area showed that the major constituents of the SFE oil in decreasing peak area were farnesol, b -gurjunene, benzyl acetate, trans-ocimene, methyl benzoate and benzyl benzoate. The major constituents of the hydrodistilled oil were benzyl acetate, p-methylanisole, trans-ocimene, methyl benzoate and farnesol.

Sensory evaluation of the two oils showed that at 1% level of significance the scent of the SFE oil was not significantly different from the scent of the fresh flowers. Moreover, the SFE oil had the most floral and sweet scent while the hydrodistilled oil had the most musky and medicinal odor.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TS1 NP3

Application of Thermal Desorption-Gas Chromatography for the Analysis of the Volatile Organic Compounds from Fresh and Dried Leaves of Vitex negundo and Psidium guajava

Fabian Dayrit Maria Cristina Dancel Denise Teotico

Department of Chemistry School of Science and Engineering, Ateneo de Manila University

Loyola Heights, Quezon City

Herbal medicine has maintained its popularity in a number of Asian countries. Because of the increasing demand for these herbal preparations, there is a need for analytical methods to determine the quality of these drugs. One way is through the analysis of volatile organic compounds (VOC’s). Analysis of VOC’s in natural products is traditionally involves steam distillation and collection via a Clevenger set up. An alternative method is dynamic headspace methods, specifically using Thermal Desorption.

The VOC’s from fresh and dried leaves of Vitex negundo and Psidium guajava were analyzed using these two methods and their results compared. Identification of the components of the VOC’s was carried out by thermal desorption-GC-MS. The effect of changing the interface temperature in the TD set-up to obtain optimum results was also examined.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TS2 NP4

ISOLATION AND CHARACTERIZATION OF TIHE PRINCIPLE IN DRimys piPeRita Hook f. (SAPAL) THAT COULD LEAD TO THE SYMPTOMATIC RELIEF OF DIARRHEA

Louisa P. Pladio1 and Irene M. Villaseņor2

1Benguet State University, La Trinidad, Benguet

2Institute of Chemistry, University of the Philippines, Diliman

Drimys piperita Hook f. locally known as "Sapal" to the Igorots of the Mountain Province, is a plant popular for its medicinal use. The dried fruits and a decoction of the leaves are believed to counter diarrhea and stomach disorders. Drimys piperita Hook f. belongs to the Winteracea family. It is a shrub, sometimes scrambling or epiphytic, or treelet, that grows from 0.1 to 13.0 m high. The fruits are obovoid to ellipsoid, nitidous and black to orange-brown and dull. The leaves are scattered to pseudoverticillate, blades are elliptic or obovate to lanceolate.

Fresh young leaves of sapal were collected from Mt. Data, Bauko, Mountain Province. The leaves were air dried and then extracted with distilled ethanol. The extract was concentrated under reduced pressure using a rota-evaporator with bath temperature maintained at 40oC. The dark green resinous residue obtained was partitioned using solvents of varying polarity: hexane, ethylacetate, and methanol. The concentrated hexane fraction was subjected to repeated rapid column chromatography applying gradient elution technique.

The bioassay-guided extraction and isolation scheme afforded two isolates. The activity of the various fractions obtained was monitored using the Charcoal Tracing Method. In this assay, Swiss albino mice were used as test animals with loperamide (immodium tablet) and normal saline solution (nss) as the positive and negative control, respectively. The extracts and isolates from the leaves of Drimys piperita Hook. f. were found to effectively control the symptoms of non-infectious castor oil-induced diarrhea in Swiss albino mice. Isolate FAC gave 99.87% inhibition of the intestinal propulsion of the charcoal meal fed to the test animals at a dosage of 2.0 mg per 20 g mouse, while isolate FC61 gave an 84.79% inhibition at a dosage of 0.80 mg per 20 g mouse. These values were comparable to the 99.7% inhibition exhibited by loperamide.

Statistical analysis using student t-test showed that both FAC and FC61 isolates were comparable to loperamide. The calculated t-values, 0.6538 and 2.0364, were not significantly different at both 1% and 5% level of significance. When compared to the normal saline solution, the calculated t-values obtained for both isolates showed a highly significant difference at both levels of significance. These results showed that FAC and FC61 are positive towards the symptomatic relief of non-infectious castor oil-induced diarrhea.

Spectral analyses using IR, 1H NMR, 13C NMR, and Mass spectroscopy showed isolate FAC to be a symmetrical 29-carbon 2o alcohol. The white powder was identified as 15-nonacosanol, CH3(CH2)13CH(OH)-(CH2)13CH3. Based on available IR, 1H NMR and 13C NMR spectral analyses, isolate FC61 which is a yellow-green oily substance is possibly an aromatic ester with a long chain alcohol portion. A mass spectral analysis is needed to confirm the molecular formula and structure of this active compound.

 

 

 

 

TS2 NP5

CHEMICAL CONSTITUENTS OF SOME PHILIPPINE BASIDIOMYCETES FUNGI

Shirley Caballero1, Adam McCluskey2 and Anamy Ma. Paano

1Chemistry Department, De La Salle University, Taft Avenue, Manila

2Chemistry Department, University of New Castle, NSW, Australia

Crude alcoholic extracts were obtained from five selected mushroom species. Crude extracts, semi-pure and pure isolates obtained by chromatography were tested for their physiological properties.

A lanosterol derivative was identified from Payporus sp., mannitol from Agaricus arvensis, Chlorophyllum molybdites, and Lepiota cristata. An aminofuranone compound was also isolated from Lepiota cristata. This is the first time this compound was isolated from a natural source.

Mannitol was, derivatized with different ketones namely acetone, cyclohexanone, benzophenone, and dibenzylacetone to explore the use of mannitol to synthesize potential anti-cancer compounds. The structure of other mushroom isolates will also be discussed.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TS2 NP6

Biological Activities of Sterols and Triterpenes from the Leaves of Coleus blumei Benth.

Rofe-Amor P. Obena and Amelia P. Guevara

Institute of Chemistry, University of the Philippines, Diliman

Mixtures of sterols (FB1b3) and of triterpenes (FC1F1a) were isolated from the leaves of Coleus blumei Benth, locally known as mayana, a known folklore medicine for pain. The isolates exhibited analgesic, anti-inflammatory and antimicrobial activities. Using a bioassay-guided fractionation, this was the first study conducted to evaluate the analgesic activity of mayana extract and the antifungal activity of the sterols. Also, this was the first report on the detection of campesterol, a -amyrin and b -amyrin from mayana.

GC-MS analysis showed that FB1b3 was a mixture of 6 components, of which the three major components are sterols, namely, stigmasterol, b -sitosterol, and campesterol. The sterol mixture (0.01% yield from 1 Kg dried leaves) obtained from the active column fraction FB inhibited the acetic acid-induced pain by 39.0% at a dosage of 0.05 mg/g mouse. Analgesic experiments by oral administration and intraperitoneal injection were performed to compare the effect of modes of administration and to determine the effect of dosage on analgesic activity. Intraperitoneal injection at a dosage of 0.05 mg/g mouse was found to be a more effective mode of administration. FB1b3 inhibited the carageenan-induced inflammation by 63.3% at a dosage of 0.1 mg/g mouse. It was also observed that at 1 m g/m L FB1b3 inhibited weakly the growth of both C. albicans and T. mentagrophytes as shown by the antimicrobial index of 0.2 but was inactive towards E. coli and S. aureus.

FC1F1a was identified to be a mixture of the pentacyclic triterpenes a -amyrin and b -amyrin with molecular mass of 426 g/mol and relative peak areas of 66.7% and 33.3%, respectively. The triterpene mixture (2.46 x 10-3% yield from 1 Kg dried leaves), also obtained from the active column FC fraction, inhibited the acetic acid-induced pain by 43.3% at a dosage of 0.05 mg/g mouse. It also inhibited the carageenan-induced inflammation by 30.8% at a dosage of 0.1 mg/g mouse but did not inhibit the microbial growth of E. coli, S. aureus, C. albicans and T. mentagrophytes at the concentration of 1 m g/ m L.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TS2 NP7

A BIOACTIVE LECTIN FROM GOLDE SNAIL (Pomacea SP.)

Flornia E. Merca1, Marivic S. Lacsamana1, Ruby A. Ynalez1 and Sonia Y. Jacinto2

1Institute of Chemistry, College of Arts and Sciences

University of the Philippines, Los Baņos, Laguna

2Institute of Biology, University of the Philippines, Diliman

A lectin specific for sialic acid was isolated and purified from the visceral region of golden snail (Pomacea sp.) by conventional protein isolation techniques and by affinity chromatography.

The lectin was found to be both non-blood type and non-blood group specific. It is a glycoprotein consisting of four different subunits with estimated molecular masses of 120 kD, 115 kD, 105 kDand 100 kD. Amino acid analysis of the lectin showed high amounts of glutamic acid, leucine and aspartic acid.

Biological assays conducted on the purified lectin showed that it has mitogenic activity and is cytotoxic towards human breast cancer cells.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TS4 NP8

 

BIOACTIVITY STUDIES ON TRITERPENES ISOLATED FROM CARMONA RETUSA (VAHL) MASAM. LEAVES

Irene M. Villaseņor, Arlyn P. Canlas, Karen M. Faustino, Katherine G. Plana

Institute of Chemistry, University of the Philippines, Diliman

Carmona retusa (Vahl) Masam. is a member of the Boraginaceae family and is locally known as tsaang gubat. Several biological activities are ascribed to the plant. It is one of the medicinal plants with a dosage formulation. Its pediatric tablet is recommended as an anti-colic and has undergone clinical trial phases 1 and 2.

The major constituent of C. retusa (Vahl) Masam. leaves is an intractable mixture ol triterpenes, namely a -amyrin, b -amyrin, and baurenol. This paper will report on the biological activities of these triterpenoidal mixture including their analgesic, anti-diabetic, anti-inflammatory, anti-microbial, anti-spasmodic, and anti-mutagenic activities.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TS4 NP9

LECTINS FROM TWO PHILIPPINE MEDICINAL PLANTS

Marivic S. Lacsamana, Ana Christina L. Opina, Grace Jones D. Kalaw and Flornia E. Merca

Institute of Chemistry, College of Arts and Sciences

University of the Philippines Los Baņos, Laguna

Fresh leaves and exudates therefrom of some plants have been used as popular remedies for a variety of unrelated human afflictions. Among these is the leaf extract of aloe plants which has been used in folk medicine as a topical treatment for skin and other cutaneous wounds and burns. Previous work on aloe leaf extracts reported the isolation and purification of lectins in these extracts. In vitro studies on these lectins showed their possible participation in the wound healing process. The Philippines abounds in plants whose leaf extracts were found to have the same medicinal action as the aloe leaf extract. It is possible that the leaves of these plants also contain lectins.

This paper reports on the identification, purification and partial characterization of two novel lectins from the leaves of two locally available medicinal plants namely. Pithecellobium dulce (Roxb.) Benth and Schefflera odorata (Blanco) Merr. & Rolfe.

The results obtained show the potential of the leaves of these two medicinal plants as readily available and cheap sources of lectins. Further studies on the biological function of these leaf lectins can shed light on their possible contribution in the reported wound healing properties of the two plants.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP1

SCREENING, ISOLATION AND PURIFICATION OF TWO LECTINS FROM MATURE LEAVES OF LARGE-LEAFED MAHOGANY SWIETENIA MACROPHYLLA KING

Abigail Joy D. Rodelas, Marivic S. Lacsamana and Florinia E. Merca

Institute of Chemistry, College of Arts and Sciences

University of the Philippines Los Baņos , Laguna

The leaves of twelve Meliaceae species were screened for the presence of lectins. Eight of these species namely, Azadirachta indica (neem), Dysoxylum cumingianum (tara-tara), Melia azedarach (chinaberrry/paraiso), Melia dubia (bagalunga), Sandoricum koeyjape (santol), Swietenia macrophylla King (large-leafed mahogany), Swietenia mahogani (small-leafed mahogany) and Toona calantas (kalantas) tested positive for lectin activity. The strongest lectin activity was observed in the crude leaf extract of S. macrophylla King.

The lectins in the mature leaves of S. macrophylla King were extracted using 0.02 M phosphate buffer containing 0.15 M NaCl, pH 7.2. Purification of the lectins was dope through sequential ammonium sulfate fractionation and gel filtration using Sephadex G-150.

The two purified lectins from the mature leaves of S. macrophylla King namely, Lectin 1 and Lectin 2, are both non-blood type specific because they agglutinated all human blood types (A, B, O and AB). SDS-PAGE gave two protein bands for Lectin 1 with estimated molecular weights of 210, and 200 kD. For Lectin 2, only one protein hand was observed with a molecular weight of approximately 202 kD.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP2

MONITORING OF THE BIOACTIVE CONSTITUENTS IN IPOMOEA MURICATA (JACQ.) CONVOLVULACEAE SEEDS COLLECTED FROM 1985-2000 BY HPTLC

  1.  
  2. G. Chan, X. L. Liu, D. S. Nepomuceno and M. C. Ysrael

College of Pharmacy, University of Santo Tomas, Espaņa, Manila

The antibacterial antifungal and analgesic in I. muricata seeds have been previously identified. However, these constituents may disappear through years of storage and may also vary from one batch collection to another. The HPTLC was viewed as a convenient means in establishing a reference fingerprint of I. muricata seeds against which raw materials for its preparation as crude drug can be evaluated. The appropriate solvent systems to effectively separate as many components as possible were determined. With indolidizidine alkaloids, prenylpropanoids and muricatin as marker compounds, the TLC profiles of the different batch collection of seeds were compared. Quantitative determination of the marker compounds was made by densitometry.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP3

PARTIAL CHARACTERIZATION OF AN ANTITUMOR COMPONENT OF BATAW

Cesar V. Ortinero, May Arcelie V. Tirante, Romina B. Dizon, Sharon E. Lazaro

Department of Chemistry, Central Luzon State University, Science City of Muņoz

A potential antitumor component of bataw (Dolichos lablab Linn) seed was isolated following a brine shrimp lethality guided fractionation and isolation scheme. The potential of the isolate was evaluated using the procedure developed by McLaughlin et al. (1993). Partial characterization of the isolate was done using IR spectroscopy.

Results of the antitumor prescreening test showed that the LC50 of the isolate against brine shrimp is 8.02 m g/mL. Since the LC50 of the isolate is less than 30 m g/mL, it is considered a potential antitumor agent. The US National Cancer Institute protocol set 30 m g/mL as the highest LC50 value a substance must have to be considered as having antitumor property.

The IR spectrum of the isolate revealed the presence of -OH, C-H, C=C and (cis) HC=CH groups in the molecule.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP4

ANTIHYPERTENSIVE TETRANORTRITERPENES AND OTHER METABOLITES FPOM SWIETENIA MACROPHYLLA KING

Arlene Sy 1, Pilar Lim-Navarro2 and Anamy Ma. Paano1

1Chemistry Department, De Lasalle University, Taft Ave. Manila

2Department of Medical Affairs, United Laboratories, USA

The hypertensive properties of various crude extracts of kernels and seed coats of Mahoqany macrophylla were previously reported. We now report the structure elucidation of the pure isolates obtained from chloroform extracts of the cotyledon and of the seed coats of mahogany seeds.

The structures of the pure isolates obtained were elucidated using literature data, melting point of the compound and the mass spectral fragmentation pattern of the compound. The structures obtained from MS data were confirmed from their 1H NMR and 13C NMR data. Four tetranortriterpenoids were identified from both the kernel and the seed coat samples, swietenine being common to both.

The antihypertensive activity of some of the pure and semi-pure isolates were also determined using spontaneously hypertensive rats (SHR) as test animals. A polygraph was used to monitor the rats’ blood pressure. Two pure isolates from the seed coat namely, swietenolide and swietenine showed significant antihypertensive activity at different doses. Swietenolide was also found to be non-toxic at doses tested.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP5

Antimicrobial Compounds from Anacardium occidentale

 

Consolacion Y. Ragasa1, Rosemarie Dumag1 and John A. Rideout2

1Chemistry Department, De La Salle University, Taft Avenue, Manila

2Chemistry Department, Central Queensland University, Rockhampton, Queensland, Australia

Anacardium occidentale afforded a mixture of 1 and 2 in an approximately 2:3 ratio and another mixture of 3 and 4 in an approximately 2:3 ratio. Their structures were elucidated by extensive 1D and 2D NMR and mass spectrometry. The mixture of 1 and 2 indicated slight activity against P. aeruginosa, C. albicans, and T. mentagrophytes, moderate activity against B. subtilis and high activity against A. niger. The mixture of 3 and 4 indicated slight activity against E. coli and P. aeruginosa and moderate activity against S. aureus, B. subtilis, C albicans, T. mentagrophytes and A. niger.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP6

STUDIES ON BIOCTIVE COMPOUNDS FROM PHILIPPINE SPONGES: ISOLATION AND CHARACTERIZATION OF ANTIMICROBIAL METABOLITES

 

Evamarie P. Capareda, Vladimir M. Esposo and Oliver B. Balagtas

Division of Organic Chemistry and Natural Products, Institute of Chemistry

University of the Philippines, Los Baņos, Laguna

Bioassay guided chromatographic separation was employed to obtain antimicrobial isolates from the extracts of two sponges collected in Palawan.

From the dichloromethane extract of the sponge Monanchora clathrata, a light green oil was obtained which displayed activity against E. coli and S. aureus in the agar plate disc diffusion assay. IR and UV analysis of the isolate suggested the presence of ester and olefinic groups.

A rusty orange Verongid sponge collected in Palawan yielded a dark yellow isolate which exhibited marked activity against S. aureus and B. subtilis, with weaker activity against E. coli, P. vulgaris and C. albicans. Spectroscopic data (IR, 1H NMR, UV) indicated the presence of OH and OR groups and conjugated unsaturation.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP7

RECOVERY AND CHARACTERIZATION OF A WATER-SOLUBLE GUM FROM SEED OF PARADISE FLOWER (CAESALPINIA PULCHERRIMA LINN.)

Rodel T. Botio

Tarlac State University, Tarlac City

Gum from the seeds of Caesalpinia pulcherrima Linn. Was recovered in various techniques namely: manual separation, fast roasting and acid decoating techniques. The average breakdown composition of pods and seeds were determined. The average physical breakdown composition of a pod of Caesalpinia pulcherrima Linn. Were: 54.0% (3.62 g) cover and 46% (2.99 g) for seeds. The average physical breakdown composition of seeds were :25.75% (0.77 g) seed coat, 35.45% (1.06 g) gum; and 38.80% (1.16 g) endosperm.

Physicochemical and chemical identification showed 88.48% Gum content and the presence of galactomannans and galactoglucomannans sugars. The Chemical characteristics of gum from the seeds of Paradise flower exhibited similar properties of commercial galactomannans containing gum.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP8

Development of Quality Assurance Methods for Medicinal Plants Using Thin Layer Chromatography Analysis

Fabian M. Dayrit and Marites J. Pasuelo

Department of Chemistry, Ateneo de Manila University, Loyola Heights, Quezon

This study demonstrates the use of thin layer chromatography (TLC) for monitoring product quality. The method makes use of Blumea balsamifera Linn. (sambong), as an example. The Retention factor (Rf) values and density of the spots were measured by scanning the plate with a flatbed scanner and quantified using the software QuantiScanŌ of Biosoft. By calculating the relative retention time (rRf) and relative peak areas (rPA) of the compounds, a template of the chromatographic profile was developed. This template can be used as a standard against which other sambong samples can be compared.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP9

Studies on the Existence of Dynophysis sp. and Gambierdiscus sp. in Philippine Coastal Waters

M. C. G. Floresca, F. F. Fajardo, J. L. V. Nicdao, K. P. Yturzaeta,

J. P. Co and J. T. Pakingan

Department of Chemistry, De La Salle University, Taft Avenue, Manila

Paralytic shellfish poisoning (PSP) has been reported in the Philippines and extensively in other parts of Asia. Currently in the Philippines, however, there are still very little or no document about PSP and its causal organisms. The present study aims to determine the presence of some of the causal organisms of PSP in the Philippines. This was done by examination of the cell morphology of the suspected causal organisms and by isolation of the toxins associated with these organisms. Samples for analysis were gathered from suspected red tide areas in the country. They were shellfish from Masbate, shellfish and fish from Romblon and water samples from Pangasinan. Extraction of the toxins was done in the following manner. Fish viscera and shellfish meat were ground using an osterizer. These samples were then cooked in acetone at 70oC then later cooled down to room temperature. A portion of the acetone layer was used in mouse bioassay while the remaining portion was purified by mixing it with diethyl ether:ethanol-water (1:3) solution and separated using a separatory funnel. The resulting acetone layer was later purified by freezing it at -20oC. The frozen (solid) layer was discarded and the liquid portion was used for vacuum liquid chromatography (VLC) with 9:1 chloroform:methanol as the mobile phase. Finally, after VLC, high performance liquid chromatography coupled with a mass spectrometer is used to elucidate the composition of the toxin.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP10

Use of Solid Phase Extraction (SPE) Techniques as Diagnostic Tools in the Chemical Characterization of Factors in Tripneustes gratiLla (Echinodermata: Echinoidea) Larval Settlement: Initial Results

M. N. E. Montaņo, S. J. P. Caņete and M. A. J. Meņes

Marine Science Institute, University of the Philippines, Diliman

Studies have shown that the sea urchin larvae completely settled and metamorphosized in Sargassum (brown seaweed) conditioned seawater. This implies that the settling factors are most likely metabolites from the seaweed. This study illustrates the use of Solid Phase Extraction (SPE) techniques, coupled with a bioassay, as a diagnostic tool to assist in the characterization and in the design of strategies to isolate the chemical factors involved in the induction of larval settlement and metamorphosis in Tripneustes gratilla (Echnordermata: Echinoidea). Through SPE, the size, charge and adsorptivity of the bioactive molecules can be obtained. Characterization and isolation of the settling factors are important in the development of hatchery techniques in the mariculture of sea urchins.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP11

CORRELATION BETWEEN THE MOUSE ACUTE ORAL LD50 AND THE BRINE SHRIMP LC50 OF SOME COMMON MEDICINAL PLANTS

Lunesa C, Pinzon, Rey Y. Capangpangan and Jefferson C. Abian

Department of Chemistry Mindanao State University- Iligan Institute of Technology, Iligan City

Thirteen (13) common medicinal plants were studied for their brine shrimp acute median lethal concentration (LC50 ) after 24 hours. These plants had been previously reported to have antibacterial or antifungal activities and with mouse acute oral median lethal dose (LD50) values varying from 38 to more than 300 g/kg body weight. The results showed that the LC50 values of the plants were in the range between 4 and 35 ppm and that, at 10 ppm concentration, the % mortality of brine shrimps was between 34 and 68%.

The study showed that all natural and synthetic materials with LC50 between 4 and 35 ppm could also be as toxic and/or bioactive as the sampled plants. It also showed that the Brine Shrimp Toxicity Test (BSTT) at 10 ppm which required only 0.25 mg of sample and 24 hours could be a reliable, fast, inexpensive, and practical substitute for the more commonly employed antibacterial and antifungal bioassays as guides in the fractionation of natural product., With BSTT as a fractionation monitor, other bioactive substances (anti- viral, anti-malarial, anti-tumor, anti-cancer, anti-AIDS, pesticidals, relaxants, depressants, etc.) might not be eliminated early in the isolation of natural products.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP12

THE INHIBITORY EFFECT OF THE AQUEOUS DECOCTION OF THE LEAVES OF Plantago major Linn (LANTING) ON THE BONE MARROW GENOTOXICITY OF TETRACYCLINE AND CHLORAMPHENICOL

Eden C. Agdon, Marie Christy M. Caringal and Jacelyn R. Castillo

College of Science, Pablo Borbon Memorial Institute of Technology, Batangas City

This study dealt mainly on the analysis of the antigenotoxic activity of Plantago major Linn (Lanting) against two genotoxins – tetracycline and chloramphenicol.

Lanting leaves were extracted by decoction. The antigenotoxic potential of lanting leaves was investigated using the Micronucleus test. Utilizing this method, two positive control groups (one in the presence of tetracycline alone and one in the presence of chloramphenicol alone) and one negative control group (distilled water) were considered as basis of comparison with the treated groups.

The incidence of micronucleated polychromatic and normochromatic erythrocytes were counted per 1,000 PCE and NCE, respectively, under a Carl-Zeiss photomicroscope using high magnification at oil immersion. The functional groups present in the aqueous decoction of lanting leaves were analyzed using Infrared Spectroscopy.

The mean incidence of MPCE obtained from group treated with tetracycline/chloramphenicol (positive control groups) is very much higher than the mean incidence of MPCE from group treated with distilled water (negative control). The mean incidence of MPCE obtained from the groups treated with tetracycline/chloramphenicol plus aqueous decoction of lanting leaves (treated groups) is relatively higher than the mean incidence of MPCE from the group treated with distilled water (negative control).

The mean incidence of MNCE obtained from group treated with tetracycline/chloramphenicol (positive control groups) is very much higher than the mean incidence of MNCE from group treated with distilled water (negative control). The mean incidence of MNCE obtained from the groups treated with tetracycline/chloramphenicol plus aqueous decoction of lanting leaves (treated groups) is relatively higher than the mean incidence of MNCE from the group treated with distilled water (negative control).

The percentage reduction in the number of MPCE in the positive control involving chloramphenicol alone yielded a greater percentage reduction than the group treated with tetracycline alone. The percentage reduction in the number of MNCE in the presence of tetracycline alone (positive control) gave a higher percentage reduction than in the treatment of chloramphenicol alone (positive control). The antigenotoxic effect exhibited by the aqueous decoction of lanting leaves on mice with tetracycline/chloramphenicol, differ significantly from that of the distilled water (negative control), and from that of the tetracycline/chloramphenicol (positive control groups) in terms of micronucleated polychromatic and micronucleated normochromatic erythrocytes. Thus, this indicates high antigenotoxicity effect exhibited by the aqueous decoction of lanting leaves. The functional groups that are inferred to be present in the aqueous decoction of lanting leaves are the following: at frequency 1640.64 cm-1, C=O; at 2958.65 cm-1, CH2; at 3411.95 cm-1 , O-H; and at 1088.45 cm-1, C-O.

 

NP13

FINGERPRINTING OF PANDANUS AMARYLLIFOLIUS ALKALOIDS: A PRELIMINARY BIOGENETIC STUDY

Daisy Lopez1 and Maribel Nonato2

1Graduate School, University of Santo Tomas, Espaņa, Manila

2College of Science and Research Center for the Natural Sciences

University of Santo Tomas Espaņa, Manila

Mature leaves of Marikina grown Pandanus amaryllifolius Roxb. collected quarterly throughout the year 1999 yield crude alkaloid fractions of similar TLC profile. Alkaloids were detected in the dichloremethane and n-butanol fractions obtained after extraction with solvents of different polarity. The dichloromethane fraction from each collection, after series of chromatographic separation, yields the alkaloids identified as Pandamarilatone-1, Pandanamine and Pandamarilactonine - A, - B and --C. Pandanamine is predicted biogenetically to be the possible intermediate of the other alkaloids and those reported in literature. The isolation and the identification of the alkaloids and their relationship with each other will be reported.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP14

A BIOACTIVE COMPOUND FROM EQUISETUM RAMOSSISSIMUM DESF. SPP.

DEBILE (ROXB) HAUKE

Ofelia DLC Giron1 and Iqbal Choudhary2

1UP College of Baguio, Baguio City

2HEJ Research Institute of Chemistry, University of Karachi, Pakistan

 

The utilization of plants for medicinal purposes is prevalent among communities in the Cordillera. One of the medicinal herbs found to be effective against kidney diroders froma survery of residents in Busol, La Trinidad and Baguio City is Equisetum ramossissimum Desf. Spp. Debile (Roxb) Hauke. The herb belongs to the family Equisateceae and is popularly known as ‘puputtod’ among the Ibalois and Kankanaeys. ‘Puputtod’ is a rough textured herb 30 cm to about 1 m tall that is commonly encountered along irrigation canals and embankments of rice terraces. It is rather common and widespread in the Philippines.

Samples of ‘puputtod’ were collected in Busol watershed and air-dried for 48 h. The plant materials were separately extracted with petroleum ether and chloroform and the resulting crude organic extracts were purified by column chromatography with silica gel and/or flash chromatography with varying gradients of polar to non-polar solvents.

The crude petroleum ether (PE) and Chloroform (CHCl3) extracts and column chromatography (CC) fractions from subsequent purification of the two extracts were investigated for their antifungal activity and toxicity to brine shrimps. The in vitro fungicidal bioassay involved four (4) human pathogens, three (3) animal pathogens and one (1) plant pathogen. The results indicate 50% inhibition for the human pathogen Aspergillus niger and the animal pathogen Microsporum canis with respect to the PE extract. The brine shrimp bioassay was aimed at screening both extracts and CC fractions for substances that are toxic to zoological systems. It was observed that the PE extract and almost all of the CC fractions from the PE extract exhibited high toxicity to Artemia salina.

Further purification of the PE extract afforded a white solid. Preliminary structural studies on the white material suggest a long chain a ,w -dicarboxylic acid.