International Journal of

Pharmaceutical and Clinical Research

e-ISSN: 0975 1556

p-ISSN 2820-2643

Peer Review Journal

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Abstract about Conference [link to conference]
To facilitate the experts meeting and sharing knowledge, Faculty of Pharmacy Gadjah Mada University has organized The 4th International Conference on Pharmacy and Advanced Pharmaceutical Sciences (ICPAPS 2015) with the captivating theme : “Integrating Sosio-entrepreneurship in Marine Development for Sustainable Pharmacy” as a response to the increasing needs for marine pharmaceuticals.The conference was held in collaboration with the Indonesian Ministry of Coordinating Maritime Affairs, Nara Institute of Technology Japan, Leiden University and the Netherland, Mahidol University Thailand and Monash University Malaysia.

1. Immunomodulatory Effect of Massoia Bark Extract and the Cytotoxicity Activity Against Fibroblast and Vero Cells in Vitro
Permanasari P , Hertiani T , Yuswanto A 
Background: Bark of Massoia aromatica Becc (Lauraceae) has been widely used in Java, Bali and Sumatra as a fragrance ingredient, seasoning and traditional medicine. C-10 Massoia lactone is a major constituent of its essential oil which has been proven as the antibacterial principle and has the ability to increase the phagocytic index of mice macrophage. Objective: to find out the immunomodulatory effects of the Massoia aromatica bark extract against Candida albicans as well as to explore the potential toxicities against fibroblast and vero cells. Material and methods: The extract was obtained by maseration of the pulverized barks with ethanol 96%, followed by solvent evaporation. The Massoia lactone contain was analysed by TLC-Densitometry. The immunological assay was based on the effect on SD mice lymphocyte proliferation and macrophage phagocytosis activities against latex and Candida albicans. Samples concentration tested were in the range of 30 – 2.5 μg/mL and done in triplicate. Cytotoxicity assays were done on fibroblast and vero cells by using the MTT assay. ANOVA followed by Tukey-HSD was used for statistical analyses. Results: The ethanolic extract was separated into two layers, i.e. solid and oily phases. All samples exhibited no effect on lymphocyte cell proliferation which indicated by the stimulation index proliferation value of less than 2. However, all samples showed activation of the macrophage phagocytosis both on latex and Candida albicans in a concentration dependant manner. IC50s of oily and solid extracts on fibroblast were 46.13 and 53.03 μg/ml respectively, while on Vero cell were found at 43.59 and 153.05 μg/ml, respectively.

2. Leunca (Solanum Nigrum L.) Herb Sensitizes Doxorubicin Resistant MCF-7 Cells
Riris Istighfari Jenie, Rumiyati, Woro Anindito Sri Tunjung, Dyaningtyas Dewi Pamungkas Putri, Adam Hermawan, Edy Meiyanto
Chemotherapeutic agent resistance is becoming a heavy obstacle in cancer therapy as it increases the toxicity of the drug to normal cells and reduces the efficacy of the drug to cancer cells. The molecular mechanism of resistance may involve Pgp protein, a transmembrane protein which has been known to induce efflux of its substrates, including chemotherapeutic drugs, from the cells. One idea to overcome this problem is by combining the chemotherapeutic agent with other substance in order to sensitize cancer cells. Leunca (Solanum nigrum L.) is a potential source of natural anticancer agent. Leunca ethanolic extract (LHE) has cytotoxic activity in several cancer cell lines such as HepG2 and HT-29. The aims of this research were to investigate the effect of leunca ethanolic extract (LHE) in combination with a chemotherapeutic agent, doxorubicin, on MCF-7/doxorubicin resistant (MCF7/DOX) cell line viability and observing the expression level of Pgp protein after combination treatment. Cell viability assay of LHE and its combination with doxorubicin were conducted using MTT [3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide] assay. Pgp expression levels were observed using immunocytochemistry. LHE showed cytotoxic effect on MCF-7/DOX cell line with IC50 344 μg/ml. Combination of 0.375 (1/16 IC50) and 0.75 μM (1/8 IC50) doxorubicin with 21.5 (1/16 IC50) and 43 μg/ml (1/8 IC50) LHE showed strong synergistic effect (CI is 0.1-0.3). The immunocytochemistry results showed that the combination treatment reduced the expression level of Pgp while the doxorubicin treatment increased Pgp expression level. These results suggested that LHE is potential to be developed as co-chemotherapeutic agent on breast cancer cells.

3. Cytotoxic and Apoptotic Effects of Para-Hydroxy Meta Methoxy Chalcone (Phmmc) on T47d Breast Cancer Cells
Retno Arianingrum, Retno Sunarminingsih, Edy Meiyanto, and Sofia Mubarika
para-Hydroxy meta methoxy chalcone (pHmMC), a chalcone derivate of 3 – (4′-hydroxy-3′-methoxyphenyl)-1-phenyl-2-propene-1-on, was investigated on its cytotoxicity, apoptosis induction, and Bcl-2 expression against human breast cancer-T47D cell lines. The cytotoxic effect was analysed using MTT [3-(4,5 dimethyltiazol-2-yl)-2,5-diphenyltetrazoilium bromide] assay, the apoptotic effect was determined by Annexin-V flowcytometry method, and the expression of Bcl-2 was identified using immunocytochemistry method.  The research results showed that the pHmMC’s IC50 on T47D cells was 48 M, and it increased the apoptotic induction and decreased the Bcl-2 expression level.

4. Antiplasmodial Activity of Endophytic Fungi Isolated from Artemisia Annua, l.
Indah Purwantini, Wahyono, Mustofa, Ratna A. Susidarti, Eti N. Sholikhah,  Rani A.N. Hestiyani
Objective: This research will observed the kinetics of fungi’s growth and the ability of endophytic fungi IP-2 and IP-6 to inhibit the growth of Plasmodium falciparum strain FCR-3. Methods: IP-2 and IP-6 were fermented in Potato Dextrose Broth (PDB) medium for 14 days at room temperature and sampling was done every 24 hours to obtain biomass. Sspecific growth rate (µ) was estimated from biomass during logarithmic phase and compared for different microorganisms. The extract of medium fermentation was obtained by extracted using ethyl acetate and then evaporated untill sticky in its consistency. The extracts were subjected to antiplasmodial activity using continuous culture of P. falciparum method which developed by Trager and Jensen. Upon reaching the desired parasitaemia, the culture was synchronized refer to Lambros and Vanderberg method. Antiplasmodial assay was performed for 72 hours. At the end of incubation, parasitaemia was determined using Giemsa-stained thin blood smears and observed using microscope. The potency of antiplasmodial activity was expressed as IC50 which calculated by probit analysis. Results: Result showed that specific growth rate for IP-2 and IP-6 were 0.04 h-1 and 0.01 h-1. Both endophytic fungi had activity as an antiplasmodial with IC50 values 42.58±5.65 and 30.08±4.73 µg/mL respectively. Conclusion: IP-2 showed faster growth rate than IP-6. At 168 hours of fermentation, growth rate of IP-2 entered stationary phase while IP-6 216 hours. Endophytic fungi IP-2 and IP-6 demonstrated antiplasmodial activity with a moderate level according to Gessler criteria. Both of endophytic fungi have a great potential to be further investigated.

5. Optimization of Extraction Process and Dechlorophyllation of Ethanolic Extract of Andrographis Paniculata Ness
Puspitasari, A, Pramono, S, Martono, S, Widyarini, S
In the recent development, natural product industry has utilized extracts as raw material despite crude drugs. This phenomenon is due to the simplicity of extract to formulate and standardize. In order to enhance the effectiveness and efficiency, it is necessary to optimize extraction process which produces extract with high active compound content. Andrographis paniculata has been widely used in traditional medicine and natural product industry, therefore its extraction need to be optimized. Some researches in the field of kinetic and modeling of A. paniculata soxhlet extraction and other non-conventional techniques have been reported, but the application of these methods in industrial scope is complicated.   This research determined the optimum extraction conditions for A. paniculata using maceration method. In this report   the study began with determining the effect of particle size, extraction time and solvent composition toward andrographolide content. Further optimization of the process parameters were solute to solvent ratio, temperature and stirring speed. To purify the extract from chlorophyll, electro coagulation method was used with electrodes variation (Aluminum and Copper). Best  extraction  conditions  for  particle  size, extraction  time  and  solvent concentration  were  found  to  be  < 0.6  mm,  500  min  and  96% ethanol  respectively. The univariate analysis using SPSS 22.0 for windows shows that there is   significant effect of the factor solute/solvent ratio, stirring speed and temperature of maceration. Electro coagulation results show that aluminum electrode gave better extract purification than copper electrode.

6.Study of Radical Scavenger Activity, Total Phenol and Flavonoid Contents of Artocarpus Altilis leaves Extracts
Fakhrudin, N,Khairunnisa, S.Y., Azzahra, A, Ajiningtyas, R.J
Free radicals are ions, molecules or atoms with unpaired electrons that are highly unstable and reactive against other molecules. Free radicals contributed in the development of various human disorders. In this study, we investigated the possible new sources of natural radical scavengers which could provide antioxidant mechanisms to protect against the negative effects of free radicals. Radical scavenger activity of water, ethanol and ethyl acetate extracts of A. altilis extracts were evaluated by the radical DPPH scavenging test. All extracts demonstrated radical scavenger activity in which the ethyl acetate extract demonstrated the strongest activity as a radical DPPH scavenger. As A. altilis leaves contains various phenol and flavonoid compounds, the content of phenol and flavonoid compounds were determined by colorimetric techniques using the Folin-Ciocalteu assay and AlCl3 reagent, respectively. Our investigation revealed that the ethyl acetate extract showed the highest activity (IC50: 66.52 ± 0.70 µg/mL) compared to the ethanol (IC50: 88.08 ± 5.54 µg/mL) and water (IC50: 399.85 ± 9.48 µg/mL) extracts. The ethyl acetate extract also contains the highest amount of phenol contents (6.47 ± 0.11 % gallic acid equivalent) compared to the ethanol and water extracts (5.78 ± 0.14 % and 1.02 ± 0.15, respectively). Interestingly, the total flavonoid content of the ethyl acetate extract is the lowest (0.22 ± 0.02 % quercetin equivalent) among the others (0.33 ± 0.01 % and 0.50 ± 0.01 % for ethanol and water extract, respectively). We found a strong correlation between the radical scavenger activity with the total phenol content of the extracts but not with the total flavonoid contents. This study indicated that A. altilis leaves represents a promising new source of natural antioxidants

7. Combination Effect of Ethylacetate Extract of Plectranthus Amboinicus (Lour.) Spreng. With Doxorubicin Againts Hela Cell Lines
Poppy Anjelisa Z. Hasibuan, Rosidah
The aims of the study were to investigate the growth-inhibiting mediating effect of Plectranthus amboinicus (Lour.) Spreng. ethylacetate extract (PAE) in combination therapy with doxorubicin againts HeLa cell lines, to analyzed the apoptotic induction and expression of cyclin D1, Bcl2 and COX-2 (cyclooxigenase-2) proteins of HeLa cell lines after treatment of PAE. The cytotoxicity effects were determined by using MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The effect of apoptosis on HeLa cell lines were observed by flowcytometry assay in single dose of PAE. The expression of cyclin D1, Bcl2 and COX-2 proteins of HeLa cell lines after treatment of PAE were identified by using immunocytochemistry. The result showed that PAE had strong synergistic effect with doxorubicin againts HeLa cells based on Combination Index analysis. PAE 28.66 µg/mL increases apoptotic induction. The PAE showed suppression of cyclin D1, Bcl2 and COX-2 expressions on HeLa cell lines. The results concluded that PAE could be a potential co-chemoterapeutic agent with doxorubicin on cervix cancer cells.

8. A Rapid, Simple, and Validated Rp-Hplc Method for Quantitative Analysis of Furosemide in Human Plasma
Hudan Taufiq, Endang Lukitaningsih, Sudibyo Martono
Furosemide was a loop diuretic drug with extensive use for the treatment of various indications so many copy products had been manufactured. A copy product of furosemide should pass bioequivalence requirements which need a validated analytical method, but the previous methods still too complicated. For this reason, a simple and rapid RP-HPLC method was developed and validated for quantifying furosemide in human plasma. Chromatographic separation was performed under isocratic elution on Luna Phenomenex® C18 (250 x 4.6 mm, 5 µm) column. The mobile phase was comprised of methanol, acetonitrile, and phosphate buffer 20 mM pH 3.0 (10:30:60 v/v/v) and pumped at flow rate of 1.6 mL/min. The UV detector was operated at      233 nm. Protein precipitation was conducted using methanol and propranolol was used as an internal standard. This method was validated according to the FDA guidelines. Calibration curve were linear (r = 0.9997) in the ranges of 400-1400 ng/mL. The lower limit of quantification (LLOQ) was established at 400 ng/mL with errors and RSD at this concentration did not exceed 20% and 8% respectively. The limit of detection (LOD) was found at concentration of 100 ng/mL. The within-run and between-run precision did not exceed 10%, whereas the errors did not exceed 6% at concentration above LLOQ. So the method was proved reliable for quantifying furosemide in human plasma.

9. In Silico and in Vivo Qualitative Relationships of Para-Aminophenol Analogues
Hari Purnomo, Umar Anggara Jenie, Agung Endro Nugroho, Harno Dwi Pranowo
HP2009 or 1,3 bis(p-hydroxyphenyl)urea and MH2011 or 1- (4-hydroxynaphtalene-1 -yl)-3-(p-hydroxyphenyl)urea are para-aminophenol analogues modified based on the paracetamol structure. In our previous study with acetic acid-induced writhing reflex model, these compounds exhibited analgesic effects with ED50 values of 58 and 10 mg/kgBW, respectively. The potencies were more potent than this of paracetamol (91 mg/kgBW). In the silico study, the docking scores against 6COX.PDB were -67.4556; -85.1618; and -95.396, respectively. In the study, we investigated in silico and in vivo qualitative relationships of these compounds. In the study, there was a qualitative relationship between in silico with in vivo studies. The more stable bonds were due to the increasing number of drug receptor amino acid that plays a role in the bonding. Paracetamol was fastened by 8 amino acids, whereas HP2009 and MH2011 were bound by 10 and 12 amino acids, respectively. In silico study (molecular docking PLANTS) can be used to predict the potential of a drug to other drugs in the same action mechanism.

10. Docking, Synthesis and Cytotoxicity Test on Human Breast Cancer Cell Line (t47d) of N-(Allylcarbamothioyl)Benzamide
Tri Widiandani, Lusiana Arifianti, Siswandono
We investigated a modification of the chemical structure in order to enhance the cytotoxicity test on human breast cancer cell lines. In this study, new compound had been found by reacting allylthiourea with benzoyl chloride. The molecular docking of this compound on EGFR (1M17.pdb) using MVD v5.5 showed that the Rerank Scores were lower than 5-fluorouracyl (5FU). It can be predicted that the compound has a higher biological activity. The synthesized of new compound N-(allylcarbamothioyl)benzamide (BATU) was performed through nucleophilic substitution mechanism of the Schotten Baumann method by using triethylamine as a base. The structure of the newly synthesized compound was confirmed by TLC, IR, 1H NMR, 13C NMR. The in vitro study of cytotoxicity test was evaluated on human breast cancer cell lines (T47D) using MTT assay. The result showed that this compound demonstrated more potent compared to 5-fluorouracil as the commercial anticancer drug, with respective IC50 were 56.5 μg/mL (BATU) and 132.4 μg/mL (5FU). It can be concluded that the synthesized compound can be further developed as a potential anticancer drug.

11. A Novel Reversed Phase High Performance Liquid Chromatography Method To Accurately Determine Low Concentrations Of Curcumin In Rat Plasma
Setyaningsih D, Murti YB, Martono S, Hinrichs W.L.J., Hertiani T, Fudholi A
Due to its lipophilic nature, curcumin levels in plasma are very low after oral administration, and therefore hard to detect. A number of chromatographic methods, including LC/MS have been developed. Although the LC/MS method is sensitive, the matrix effect can be difficult to handle. Furthermore, LC/MS equipment is relatively expensive compared to the conventional RP-HPLC. Therefore, the aim of this study was to develop a sensitive and reliable method for the determination of curcumin concentration in plasma using an RP-HPLC. Curcuma longa extract was used which contains three curcuminoids. The method started by selection of mobile phase to optimally separate the curcuminoids. The best mobile phase composition was used to analyze the plasma samples. Rat plasma was spiked with curcumin and processed for protein precipitation followed by liquid-liquid extraction of curcuminoids and an internal standard, emodin. Chromatogaphic separation of curcuminoids and emodin was achieved using a Knauer C-18 column (250 x 4.6 mm; particle size: 5µm) and a gradient program of mobile phase of three solvents, methanol-acetonitrile-1% acetic acid. A gradient elution was applied with increasing the ratio of the volume percentages of acetonitrile to acetic acid from 50/45 to 53/42 during 5 minute and thereafter elution was isocratic for 15 minute. The methanol concentration was kept constant at 5 vol-% during the whole run. The method was validated according to the FDA guidelines.The method was selective, with an excellent resolution (Rs value > 2.5). The peak shape of both curcumin and emodin were symmetric with a tailing factor of 0.9-1.1. The method linearity (correlation coefficient of 0.999) was demonstrated at 6 to 200 ng/mL. The intra- and inter-day precision was 5.90-8.50% and 5.37-11.26%, respectively; the intra- and inter-day accuracy was 92.47-103.61% and 96.17-105.70%, respectively. In conclusion, the RP-HPLC method meets the validation requirements as described in the FDA guidelines and is applicable to accurately quantify curcumin concentrations as low as 6 ng/mL in rat plasma samples.

12. 6-Allyl-3-(4-Methoxybenzyl-8-Methoxy-3,4-Dihydro-2h-Benzo[E][1,3]Oxazine and 4-Allyl-2-Methoxy-6-(4-Methoxybenzyl)Aminomethylphenol: Synthesis and Cytotoxicity Test on Mcf-7 Cells
Marcellino Rudyanto, Tri Widiandani, Achmad Syahrani
The aim of the present study is to synthesize new benzoxazine and aminomethyl derivatives of eugenol, and to examine the cytotoxicity of the derivatives on MCF-7 cells. 6-Alyl-3-(4-methoxybenzyl-8-methoxy-3,4-dihydro-2H-benzo[e]1,3 oxazine was prepared by Mannich Reaction between eugenol, formaldehyde, and 4-methoxybenzylamine in methanol. 4-Allyl-2-methoxy-6-(4-methoxybenzyl)aminomethylphenol was prepared by hydrolysis of the benzoxazine with hydrochloric acid in 1-propanol followed by treatment with ammonia. The obtained compounds were subjected to cytotoxicity study using MTT assay and calculate the percent of cell viability. As the results, 6-allyl-3-(4-methoxybenzyl-8-methoxy-3,4-dihydro-2H-benzo[e]1,3 oxazine was obtained in 94% yield, while 4-allyl-2-methoxy-6-(4-methoxybenzyl)aminomethylphenol was obtained in 87% yield. The obtained LC50 values of the tested compounds on MCF-7 cancer cell line are respectively 22.5±0.20  and 24.6±0.92 µg/ml.

13. Development and Validation of Hplc Method for Determination of Curcuminoids in Indonesian Herbal Drinks
Febri Annuryanti, Asri Darmawati, Juniar Moechtar
A high performance liquid chromatographic (HPLC) method for simultaneous determination of curcumin (C), demethoxycurcumin (DMC), and bis-demethoxycurcumin (BDMC) has been developed. Reversed phase C-18 column (Licrosphere) was used as a separation column with a mobile phase consisted of acetonitrile:1% trifluoroacetic acid (50:50) and flow rate of 1.0 ml/min. Elution was monitored at 422 nm. The method showed sensitivity, selectivity, precision and reproducibility at the selected HPLC conditions. Linear range was found from 60 to 200 µg / ml, with Vxo value less than 2%. The percent recoveries of curcuminoids were 110.61 ± 6.51, 107.69 ± 5.91 and 102.08 ± 4.89 % of C, DMC and BDMC, respectively. Further, the method was used for determination of curcuminoids in herbal drinks containing curcuminoids.

14. Effect of Mobile Phase Composition, Organic Modifier, and Flow Rate on Selectivity and Retention of Stevioside and Rebaudioside A on Isocratic RP-HPLC Analysis
Yohanes Martono, Sugeng Riyanto, Abdul Rohman, Sudibyo Martono
Stevioside and rebaudioside A are diterpene glycoside compounds from Stevia rebaudiana Bert. plant. These compounds are used as non-caloric natural sweetener and antidiabetic agent. Stevioside and rebaudioside A have close polarity; therefore it is a challenge to separate these compounds using isocratic RP-HPLC.  The goal of this research was to study the effect of mobile phase compositions, organic modifier, and flow rate on selectivity and retention of stevioside and rebaudiosideA on isocratic RP-HPLC analysis. The developed method was applied to determine selectivity of respected compounds in sample matrix of Stevia rebaudiana leaves. The mixture of water-metanol-acetonitrile with certain composition was optimized. Trifluoroacetic acid (TFA) as organic modifier was added. Flow rate of mobile phase was varied between 0.6 – 1.0 mL/min. The result showed that mobile phase mixture of water : methanol (90 : 10, v/v, pH = 3.0) : acetonitrile : TFA ( 65 : 35 : 0.01, v/v/v) and flow rate 0.6 mL/min could resolve stevioside and rebaudioside A compounds on isoractic RP-HPLC. The resolution (R) of these compounds was 2.36. Mobile phase compositions in small variation did not influence the retention significantly, however, flow rate of mobile phase influenced both selectivity and retention.  The proposed method was selective, efficient, and simple to analyse stevioside and rebaudioside A in complex sample matrix of Stevia rebaudiana leaves.

15. The Stereospecific Analysis of Canarium Indicum Oil-Fatty Acid Based in Triglycerides Using High-Performance Liquid Chromatography
Rahman H, Sitompul JP, Anggadiredja K, Lee HW, Gusdinar T
Canarium seeds produce vegetable oil which comprises of triglycerides with fatty acids composition. The purpose of this study was to analyze the composition and stereospecific distribution of fatty acid in triglyceride from canarium oil extracted by mechanical pressing. The total fatty acids at sn-1, sn-2 and sn-3 positions were determined after saponification, while sn-1 and sn-3 were determined after enzymatic lipolysis reaction and followed by saponification. The analysis of fatty acids was performed by high-performance liquid chromatography (HPLC). The major composition of fatty acids in canarium oil was oleic acid (51.99%), linoleic acid (32.97%), and palmitic acid (9.82%), while the total unsaturated fatty acids at sn-2 position was 63.36%. The result of this study provides a preliminary data for assessing the nutritional value of canarium oil in food applications.

16. Profile of Mycolic Acid Cleavage Products of Isoniazid Resistant Mycobacterium tuberculosis Isolate By Gas Chromatography-Mass Spectrometry
Asri Darmawati, Deby Kusumaningrum, Isnaeni, Muhamad Zainuddin
Mycolic acid cleavage products (MACPs) profile of Mycobacterium tuberculosis (MTB) which is resistant to isoniazid (INH) has been studied. This study describes the profile of MAs based on their cleavage products (MACPs) after esterification process, using BF3 in methanol as a catalyst, by gas chromatography-mass spectrometry detector (GC-MSD). The MACPs profile of INH resistant MTB was compared to MACPs of H37Rv strain and MACPs of INH sensitive MTB isolate. Objective of this study was to obtain the characteristic profile of cellular fatty acid and MACPs of INH-resistant MTB isolate using GC-MS. Samples were cultured in Middlebrook 7H10 medium for more than 21 days. MAs and cellular fatty acids were extracted with chloroform prior to esterification for 1 hour at (90-95)o C using methanolic-BF3 as a catalyst. MACPs were extracted with heptane before injected into GC-MSD. This study obtained that the profile of six characteristic peaks of cellular fatty acids and MACPs of samples of INH-resistant MTB isolate was not significantly different to the INH-sensitive MTB isolate. But, the profile of H37Rv strain was significantly different compared to both INH-resistant MTB and INH-sensitive MTB isolate.

17. In Vitro Ace Inhibitory Assay of Extracts Containing Flavonoid-Phenolic Compounds of Edible Plants
Rumiyati, Sudarsono, Susanto B.O., Mayasari S.K., Wijaya R.N
Inhibition of Angiotensin Converting Enzyme (ACE) plays an important role to treat hypertension. Flavonoid compounds have been known to have  ACE inhibitor activity. The purpose of the present study was to evaluate ACE inhibitior activity of flavonoid-phenolic fraction from extracts of three edible plants using in vitro assay. The investigated samples were Vigna unguiculata, Allium fistulosum and Manihot utilissima. Flavonoid-phenolic compounds in the extracts obtained from the samples were analyzed using Thin Layer Chromatography and Densitometer techniques. Total phenolic contents were determined by Folin-Ciocalteu colorimetric method. ACE inhibitory assay method was based on hydrolysis of substrate HHL (Hippuryl-L-Histidyl-L-Leucine) by ACE into HA (Hippuric Acid) product. The amount of HA (Hippuric Acid) represented ACE activity and it was measured  using spectrophotometer at 228 nm. Then, ACE inhibitor activity was determined by % ACE inhibitor value. The result of the present study showed that each fraction from Vigna unguiculata, Allium fistulosum and Manihot utilissima extracts contained flavonoid-phenolic compounds with total phenolic contents (per 100 g fresh weight) were 21.99± 0.64 mg GAE, 66.29±1.79 mg GAE and 113.12±1.29 mg GAE, respectively. ACE Inhibitor activity of flavonoid-phenolic fraction from Vigna unguiculata, Allium fistulosum and Manihot utilissima extracts at 100 μg/mL concentration were 23.77±9.19%, 86.75±12.44% and 105.59±12.26%, respectively. From this study, flavonoid-phenolic fractions from Allium fistulosum and Manihot utilissima extracts might be potential to develop as antihypertensive agent.

18. Use of Thin Layer Chromatography and FTIR Spectroscopy Along with Multivariate Calibration for Analysis of Individual Curcuminoid in Turmeric (Curcuma longa Linn) Powder
Abdul Rohman, Hatifah Prapti Lestari, Ratna Wulandari Muhammad Khairiskam
Fourier transform infrared (FTIR) spectroscopy in combination with multivariate calibration of partial least square (PLS) for quantitative analysis of individual curcuminoid, namely curcumin (C), demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC) in the powder of Curcuma longa Linn. C. longa Linn was compiled from several market in district of Java region has been developed. Thin layer chromatography (TLC) was used as reference methods, and the level of curcuminoids obtained by TLC method was used as actual values. During analysis using FTIR spectroscopy, some optimization in terms of selecting the best wavenumbers regions capable of providing the high coefficient of determination (R2) and low values of errors was taken. After optimization, curcumin was predicted by FTIR spectroscopy combined with partial least of square using 40 different wavenumbers and the coefficient of determination (R2) for the relationship between actual values of curcumin determined using TLC and FTIR predicted values using absorbancies at these forty wavenumbers is 0.8716. These wavenumbers were also used for prediction of BDMC having R2 of 0.9158. Finally, the R2 for the relationship between actual values of BDMC and FTIR predicted values are 0.9023. These results indicated that FTIR spectroscopy combined with PLS is an alternative technique for determination of C, DMC and BDMC in turmeric powder. The developed method (FTIR spectroscopy) is rapid, no sample preparation and not involving excessive solvents and reagens.

19. An Experimental Design Of SNEDDS Template Loaded With Bovine Serum Albumin and Optimization Using D-Optimal
Winarti, L, Martien, R, Suwaldi, Hakim, L
Self Nanoemulsifying Drug Delivery System (SNEDDS) is not only limited to develop Biopharmaceutics Classification System (BCS) class II drugs, but also BCS class III and IV. Protein is classified into BCS class III because of its good solubility and its poor permeability. It needs suitable composition to load hydrophilic substance into lipophilic carrier such as SNEDDS. The objective of the study was to optimize SNEDDS template for Bovine Serum Albumin (BSA) protein using D-optimal mixture design. In this study, BSA was used and loaded into a SNEDDS template. Mixture of D-optimal design was used for optimization. The optimized template contained minimum amount of surfactant and maximum amount of oil that shows enhanced transmittance and emulsification rate. Sixteen formulas consisting of oil phase (X1, Mygliol 812), surfactant (X2, Chremophore EL 40), and co-surfactant (X3, Span 20) were evaluated for transmittance and emulsification time. The optimum SNEDDS template was then loaded with 1mg/ml BSA. Characterization results of the optimum template showed 55,4 nm of the size, -0,9 mV of zeta potential, spheric morphology, 34,24±0,53 second of emulsifying time, 94,20±0,82% of transmittance, and 87±0,4 0C of cloud point. The template was also resistant to 100, 200 and 1000 times dilution using water as medium, Simulated Gastric Fluid (SGF), and Simulated Intestinal Fluid (SIF).

20. Application of Simplex Lattice Design for the Optimization of the Piroxicam Nanosupensions Formulation using Evaporative Antisolvent Technique
Tuti Sri Suhesti, Achmad Fudholi, Ronny Martien
The aim of this study is to develop a polymeric drug delivery system for a nonsteroid anti inflammatory. Nanosuspensions of piroxicam were prepared by evaporative antisolvent precipitation methode (bottom-up technique) proved to be effective in improving the oral bioavailability as a result of enhanced solubility and bioavailability. To achieve this objective, the formulations were prepared by using experimental design (Simplex Lattice) to study the effects of  formulation, variables as Chitosan (X1), Polivinylpirolidon PVP K30 (X2) and Sodium Tripoliposphat STPP (X3) in combination or singly on response of particle size (Zav), polidispersity index (DPI), zeta potensial (ZP) and entrapment efficiency (%EE) of nanosuspensions. The physical characteristics of nanosuspensions piroxicam were evaluated by using particle size analyzer and a UV–visible spectrophotometer. The results of optimized formulations showed that the interaction among Chitosan, PVP K30 and STPP  was the most dominant factor to influence particle size distribution, zeta potential, polydispersity index and % entrapment efficiency. The nanoprecipitation method was used to prepare biodegradable nanosuspensions of reproducible sizes in the range (100-300 nm) with spherical shape by addressing the effects of processing parameters. Based on superimpose counterplot, it is obtained the optimum area with proportion of (X1):27,5-43,4% ; (X2): 20,9-46,9%; (X3): 30,6-40,2%. The use of an experiments design allows us to study the influence of different factors of formulation and manufacturing process on the formulation process, and to reduce the number of experiments.

21. Evaluation of Anti-Fertility Effect of Aqueous Extract of Costus speciosus (Koen.) J.E. Smith Rhizome in Mice
Sari I P, Nurrochmad A, Rahayu S
Aim: to determine the antifertility effect of the rhizome Costus speciosus (Koen.) J.E.Smith aqueous extract (CSRE) in Balb/C mice. Material and methods: the dried rhizome of Costus speciosus was subjected to extraction by maceration method using water as a solvent. Dose selection for acute oral toxicity was made on the basis of acute oral toxicity study (2000mg/kg) as per OECD guidelines. Anti-fertility dose selection was made on the basis of local community members’ experience of its uses as an anti-fertility substance. The mice were divided into groups in which they were orally administered with the doses of aqueous extract (275, 550, 1100 mg/kg). Results: LD50 CSRE was more than 5000 mg/kg. Oral administration of CSRE for 14 days resulted in a significant reduction in spermatozoa level and quality. Testes histology after CSRE treatments were similar with normal group. Conclusion: The results suggest that CSRE inhibits the spermatozoa quantity and quality on male mice.

22. Blood Glucose Level and Lipid Profile of Streptozotozin-Induced Diabetic Rats Treated With Sargassum polystum Extract
Amir Husni, Fajar Panji Anggara, Alim Isnansetyo, Agung Endro Nugroho
Diabetes mellitus (DM) is a metabolic disorder with symptoms of chronic hyperglycemia and disturbances in metabolism of carbohydrates, fats and proteins due to insulin deficiency, insulin resistance or inability of β-cells of pancreas to produce insulin. This study was aimed to evaluate the effects of ethanolic extract from Sargassum polycystum on glucose level and lipid profile in streptozotocin-induced diabetic rats. S. polycystum was extracted with ethanol 96%, then the extract was tested for in vivo experiment and analyzed using GC-MS. Ethanolic extract of S. polycystum at 150 mg/kg and 450 mg/kg was administered orally to streptozotocin induced diabetic rats, once daily for 15 days. The results obtained were then compared with normal controls (non-diabetes+normal saline 0.9% p.o), negative control (diabetes +CMC-Na 0.5% p.o) and positive control (diabetes+glibenclamide 5 mg/kg p.o). The glucose levels, lipis profile and body weight were measured on normal condition (baseline), on 0, 5th, 10th, 15th day, and pancreatic histopathological study was done on 15th day. Result showed that S. polycystum extract with 150 mg/kg had the highest capability on prevention of DM indicated by the increase in body weight, decreased  of  blood glucose values, increased of cholesterol values, decreased of triglyceride values, increased of HDL-c and LDL-c. Pancreatic necrosis was found in all streptozotocin-induced rats. S. polycystum extract with 450 mg/kg concentrations showed highest reduction of  β-cells of the pancreas induced by streptozotocin is better than  S. polycystum extract with 150 mg/kg. S. polycystum extract contains terpenoids and phenolic such as camphor, elemol, β-elemene, α-curcumene, germacrone and phenol which might be potential as oral antidiabetes therapy.

23. The Effect of Tuna Fish Oil (Thunnus albacares) on the Total Cholesterol, LDL Cholesterol, HDL Cholesterol and the Triacylglycerol Level on Hypercholesterolemia Rats (Rattus norvegicus)
Tangka J, Banne Y, Dumanauw J M, Rumagit B I
Setting a daily diet is the first step in tackling the risk of CHD, which is due to the increasing levels of cholesterol and fat. This can be done by consuming fish that is a natural source of ω-3 fatty acids, EPA and DHA. One of the products processed from the extraction of fish, especially fatty fish is tuna fish oil that almost 40% of the total fat content is ω-3 PUFAs are EPA and DHA. This study aimed to determine the effect of Tuna fish oil (Thunnus albacares) against the total cholesterol, LDL cholesterol, HDL cholesterol and triacylglycerol levels in the blood of hypercholesterolemia rats (Rattus norvegicus). The fish oil was extracted from fish waste Tuna with a reflux method using chloroform,  and the levels of EPA and DHA were determined using a gas liquid chromatography. The samples were 40 male rats. Five rats were used as a normal control group, and 35 rats were subsequently given a high-cholesterol diet for 6 weeks to achieve a state of hypercholesterolemia. Five rats from the experiment group were taken as data for hypercholesterolemia. Of 35 rats, 30 were divided into 3 groups of 10 rats: 1) a control posttest group (P0), receiving the treatment of a high-cholesterol diet, 2) the first treatment group (P1), which was given a high-cholesterol diet + 1 dose (0.5 ml) of Tuna fish oil and 3) the second treatment group (P2), which was given a high-cholesterol diet + 2 doses (1 ml) of Tuna fish oil. The treatment was given for four weeks and the data were then analyzed using descriptive statistics and ANOVA test, followed by LSD test. The results showed that the Tuna fish oil supplementation can reduce the total cholesterol, the LDL cholesterol and the triacylglycerol levels significantly with p <0.05. It can also increase the HDL cholesterol levels significantly with p <0.05.

24. Antiatherosclerosis Effect of Purified Andrographis paniculata Extract
Warditiani N K, Widjaja I NK, Larasanty L P F, Noviyanti N W R, Gitarini N M, Juniari N P M, Siahaan T F, Nugroho A E, Ramona Y , Wirasuta I M A G , Pramono S
The antiatherosclerosis effect of purified Andrographis paniculata extract (PAE) was studied on atherosclerotic Wistar male albino rats. A fat-rich diet containing 10% yolk, 15% fat of pig, 1% calcium, 20,000 IU vitamin D3 was administered to albino male wistar rats for 60 days to induce atherosclerosis. The two groups atherosclerosis rats were treated orally with PAE for 30 days at doses  of 24 and 70 mg/kg body weight, respectively. Its lipid level in blood was measured at before and the last of the PAE treatments. At the end of the PAE treatments, the rats were sacrificed and the morphology of their aorta was observed under a microscope. A TLC-densitometer was used to analyze the andrographolide content in the PAE. The results showed that andrographolide was found in the PAE as it produced Rf value of 0.5 which was similar to andrographolide refference standard.  The fat-rich diet supplemented with vitamin D3 and calcium induced dyslipidemia and atherosclerosis in rats. This was indicated by accumulation of fat plaques or calcification on the wall of their arteries.  The PAE can become an antidyslipidemia by lowering total cholesterol, triglyceride and LDL-C and also increasing HDL-C in blood (p< 0.05). The PAE dose of 70 mg/kg body weight, removed lipid plaques significantly until the same level observed in normal conditions). The andrographolide content of the PAE might be responsible for the removal of lipid plaques on the wall of rat arteries.

25. Evaluation of Antihyperlipidemic Activity and Total Flavonoid Content of Artocarpus altilis Leaves Extracts
Fajaryanti N, Nurrochmad A, Fakhrudin N
Hyperlipidemic is a lipid metabolism disorder characterized by high levels of blood cholesterol, triglycerides and LDL and low HDL levels. Hyperlipidemic is one of the most important risk factor of atherosclerosis leading to cardiovascular diseases. Breadfruit (Artocarpus altilis (Parkinson) Fosberg.) is an Indonesian native plant traditionally used as a cholesterol-lowering remedy. Breadfruit leaf water extract contains various flavonoid compounds, especially prenylated and geranylated flavonoids. The aim of this study was to investigate the antihyperlipidemic effect of water, ethanol and ethyl acetate extracts of breadfruit leaf by measuring total cholesterol, triglycerides, LDL and HDL levels in male rats induced by a high-fat diet and propylthiouracil (PTU). We also examined whether the antihyperlipidemic effect correlats with the level of flavonoid. The antihiperlipidemic effect of the extracts was tested in white male Wistar rats fed with a high-fat (4 mL/200 g) and 0.01% PTU daily for 40 days consecutively. As a positive control, simvastatin (1.26 mg/kg BW) was used. The blood levels of cholesterol, triglycerides, LDL and HDL was determined using ELISA. The total flavonoid content of water, ethanol and ethyl acetate extracts was measured using spectrophotometry after addition AlCl3. We found that the water, ethanol and ethyl acetate extract of breadfruit leaves exhibits antihyperlipidemic effect by lowering total cholesterol, triglycerides, and LDL levels and increasing the level of HDL. Compare to the water and ethanol extract, the ethyl acetate extract showed the highest antihyperlipidemic activity. The decreasing of triglycerides and LDL levels and increasing of HDL level but not the decreasing of cholesterol levels upon the treatment of ethyl acetate extracts have a positive correlation with flavonoid content.

26. Cardioprotective Effect of Ethylacetate Extract of Poguntano (Picria fel-terrae Lour.) Against Doxorubicin-Induced Cardiotoxicity in Rats
Yosua Sihotang, Jansen Silalahi, Sumadio Hadisahputra, Poppy Anjelisa, Denny Satria
This study was carried out to investigate phytochemical screening of ethylacetate extract of Picria fel-terrae Lour. leaves (EEP) and cardioprotective effect against doxorubicin-induced cardiomyopathy in rats. Dry EEP was prepared from dry grounded leaves by cold maceration, and extract was given at dosage of 300 mg/kg bw, then it’s cardioprotective effects against doxorubicin (DOX) induced cardiotoxicity was evaluated. DOX was administered intraperitoneally to rats at dosage of 20 mg/bw once a day for two days. Cardioprotective effect was evaluated by measuring biomarkers troponin T (cTnT), CK-MB levels. Histopathology of rat’s heart tissue was also examined. Result of phytochemical screening of extract was found to contain flavonoids, tannins, glycosides, and saponins. Levels of cTnT and CK-MB in DOX group are significantly different from that in control group, EEP, EEP + DOX, vitamin E + DOX, and rutine + DOX (p <0.05). DOX significantly raised cTnT and CK-MB levels (p<0,05) but decreased after administration of vitamin E, rutin, and EEP. Histopathological analysis of rat’s heart tissue showed to resulted in myocytolysis with congestion of blood vessels, pyknosis, cytoplasmic vacuolization and fragmentation. Concomitant treatment with vitamin E, rutin, and EEP revealed normal muscle fiber. These results suggest that EEP has cardioprotective effect.

27. Investigation of Anti-inflammatory Activities of Curcuma aeruginosa Roxb. in Experimental Animals
Triastuti, A, Wibowo, A, Astuti, P
Curcuma aeruginosa has been used empirically for asthma, scurvy, pain, and inflammation of uterus. The present study evaluated the anti-inflammatory activity of C. aeruginosa ethanolic extracts (CAE) in experimental animals. The topical anti-inflammatory activity was analyzed using croton oil-induced ear edema. Topical application of CAE (80 mg/ear, 160 mg/ear) reduced ear edema and pro-inflammatory cells (histological analysis) in the tissue and its activity was similar to hydrocortisone 1% (p>0.05). In the thioglycollate-induced peritonitis model, CAE at doses of 200 mg/kgBW and 400 mg/kgBW significantly inhibited leukocytes migration in balb/c mice although the activity was lower than indomethacin (10 mg/kgBW). These consistently results support the notion that C. aeruginosa can be used as potent anti-inflammatory agent.

28. Validation of an Ion-Pair High Performance Liquid  Chromatography for The Determination of Pseudoephedrine HCl, Guaifenesin  and Dexchlorpheniramine Maleat in Cough and Cold Medicines
Maria Yolanda, Sudibyo Martono, Abdul Rohman
An ior-pair HPLC method has been developed and validated for simultaneous determination of pseudoephedrine HCl (PSEU), guaifenesin (GUA) and dexchlorpheniramine maleat (DEX) in cough and cold syrup. This method is based on the interaction of analits and ion-pairing agent and was achieved on the Phenomenex C18 column (250 x 4.6 mm, 5 µm) using an isocratic mobile phase consisting of methanol:acetonitrile : 10mM Na-pentansulphonate at pH 4,0±0,1 (55:5:40 v/v/v). The analysis was performed at a flow rate of 1 mL/min, using UV detection at 218 nm. The result showed good linearity over the range of 96 – 144 µg/mL for PSEU, 320 – 480 µg/mL for GUA and 6.4 – 9.6 µg/mL for DEX with correlation coefficients more than 0.999. The mean value of recovery were 100.415% for PSEU, 99.898% for GUA and 100.822 for DEX. The validated method was successfully applied to determine these compounds in two commercial cough and cold syrup without being interrupted by the presence of other excipients. Small variations in method condition did not provide a significant difference to the concentration indicated that the method is robust.

29. Potential Bioactivity of Artificially Fragmented Soft Coral Sinularia sp. and Lobophytum sp. Transplantation
Hefni Effendi
The study aimed to investigate the bioactivity of artificially fragmented Sinularia sp. and Lobophytum sp. transplanted at 3 m and 10 m seawater depth.  Artificial fragmentation and transplantation was performed at Pramuka Island, Kepulauan Seribu.  Lobophytum sp. raw extract showed higher bioactivity toward S. aureus and E. coli than that of Sinularia sp.  Bioactivity of Lobophytum sp. and Sinularia sp. at depth of 10 m indicated higher than that of 3 m depth.  Bioactivity of Lobophytum towards Artemia salina was classified as very toxic, meanwhile bioactivity of Sinularia sp was toxic.

30. Comparison of Electrolyte Disturbance of Using Intravenous Aminophylline Versus Nebulization Salbutamol for Exacerbation Asthma in Surabaya, Indonesia
Amelia Lorensia, Zullies Ikawati, Tri Murti Andayani, Daniel Maranatha, Mariana Wahjudi
Background: Uncontrolled asthma symptoms will exacerbate asthma. Aminofilin is now rarely used as asthma medication abroad because it shows major side effects, unlike in Indonesia, which is still widely used with relatively rare side effects events. Aminophylline have relatively more affordable price compared to salbutamol, the first-line option in the management of asthma exacerbations requirements. Both of these drugs have a risk of causing electrolyte disturbances, which could endanger the patients. Considering the individual drug side effects, so it important to study the safety of the medicine to ascertain whether there are differences in the incidence of electrolyte disturbances by both drugs. Objective: The main objective of this study was to determine differences in the incidence of electrolyte disturbances in patients with asthma exacerbations prescribed with aminophylline and salbutamol at a hospital in Surabaya. Method: This study design is a cross-sectional study of inpatient adult asthma exacerbations in emergency room (ER), using two groups, namely the group receiving intravenous aminophylline therapy (n = 22), and the group receiving nebulized salbutamol therapy (n = 21). Result: There is no significant difference between in intravenous aminophylline anda nebulized salbutamol group. And there is no significant difference in sodium (p>0,866) and potassium (p>0,470) level in blood as a respon to the asthma exacerbations treatment by intravenous aminophylline compared  to nebulized salbutamol. Conclusion: Although there was no significant difference in the incidence of electrolyte disturbances in both drugs, but close monitoring is still needed to prevent any side effects incident.

31. Evaluation on Pharmacotherapy Learning Strategies Based on Reflective Pedagogy Paradigm in Pharmacist Education Program Sanata Dharma University
Yosef Wijoyo, Gandes Retno Rahayu, Iwan Dwiprahasto
Background: Reflective Pedagogigal Paradigm offers possibility to strengthen capability of student to apply their competence in real setting when they graduate. Aim: The purpose of this study is to determine: (1) the effect on students’ competence and communication skills after the application of learning strategies based on RPP; and (2) the correlation between the ability of the student reflection and the students’ competence and communication skills. Method: The study population included students in the faculty of Pharmacy, University of Sanata Dharma with interest in Community Clinical Pharmacy(CCP). The study design used was quasi-experimental non equivalent (pre-test post-test) control group design. The data was obtained from MCQs-test and role play observation for control and intervention groups and student journal reflection and diary for intervention group only. We analyzed the data quantitatively. Results: The results showed that (1) during the pre test the competence and communication skills in intervention group was 6.06 ± 0.98; while in control group was 6.24 ± 0.60 (p = 0.261), during the post test the competence and communication skills in intervention group was 7.34 ± 0.63; while in control group was 6.81 ± 0.66 (p=0.0004) (2) abilities of students’ reflection in intervention group were increasing in the last 4 weeks over 6 weeks treatment,  and (3) correlation between student reflection and students’ competence and communication skills was 0.308 (r table = 0.339). The study also found that intervention group showed capability to play roles as care giver, communicator, life-long learner and decision maker, as part of the core character building in becoming seven stars pharmacists. Conclusion: The implementation of RPP-learning strategies significantly increases the students’ competence and communication skill and ability to write reflection. There is no correlation between student reflection and students’ competence and communication skills. Application of RPP also stimulates students to build core characters qualities of seven stars pharmacist.

32. Formulation and Characterization Insulin Nanoparticle Using Low Molecular Weight Chitosan and Pectin Polymers with Ionic Gelation Method
Martien R, Sa’adah N, Saifullah T N S
Applications insulin orally as an alternative to insulin injections, encounter obstacles such as degradation by protease enzymes and insulin poor permeability in the gastrointestinal tract. One solution offered is insulin formulation into the nanoparticles form. In this study, nanoparticle formulation of insulin used polymers combination of low molecular weight chitosan and pectin. The aim of this study was to obtain the optimum formula of insulin nanoparticles that can be used as an alternative therapy for patient with DM. Preparation of insulin nanoparticles was carried out by ionic gelation method utilizing polyelectrolyte interaction between (-NH3+) of chitosan and (-COO) of pectin to form nanoparticles that are compact and stable charge. Formula optimization was performed using Factorial Design 22 with Design Expert® 7.1.5 software. Concentration and pH of the pectin were used as factors, while the entrapment efficiency, particle size and polydispersity index were used as responses. The optimum formula was further evaluated like zeta potential, particle morphology, profile spectra of FT-IR and in vitro release study. The obtained optimum formula consist of chitosan of 0.05% and 0.4% pectin (pH 5.0) with the mean entrapment efficiency of 63,59% ± 2,17, particle size of 228,3 nm ± 26,3, polydispersity index of 0,354 ± 0,042, zeta potential 49,40 mV ± 11,59, particle round and dark, ionic gelation was confirmed by FT-IR and the release profile following the kinetics Korsmeyer-Peppas models with Fickian diffusion release mechanism on media HCl buffer pH 1,2 (n = 0,363) and (n = 0,318) on PBS buffer pH 6,8 were analyzed by DDSolver program.

33. The Appropriateness of NSAIDs Utilization in a Community Pharmacy at Medan, Indonesia
Tanjung, H R, Sarriff, A, Harahap, U
Non Steroidal Anti-Inflammatory Drugs (NSAIDs) were including the most often drug involved in The Drug Therapy Problems incident. The high use of NSAIDs viewed as an important public health problem around the world. This was an exploratory and descriptive study aimed to identify the appropriateness of prescribed NSAIDs utilization and its impact on therapeutic outcome. The appropriateness of NSAIDs utilization was measured using the Medication Appropriatenes Index (MAI). This study was conducted in a community pharmacy setting at Medan, Indonesia. The duration of the study was from July 2009 to October 2010. It was shown that from 135 patients who participated, the average of MAI score was 1.45±1.23 and 21.6% has 3 or more MAI score. The statistical analysis result showed that MAI score has a significant correlation with the patient’s therapeutic outcome (p<0.05). It concluded that the appropriateness of prescribed NSAIDs utilization in the study was high. However, this high appropriateness did not appear to have strong correlations with therapeutic outcome (r = -0.238, p< 0.05) due to the therapeutic outcome of the patient’s that used NSAIDs mostly related to with effectiveness and the presence of adverse drug reaction that did not address in MAI tool.

34. Polyketide Isolation from Anona muricata Linn Leaves Enhance  p53 Expression on Raji Cell Line
Artanti A N, Astirin O P, Prayitno A
Objective: One of the compounds found effication as an anticancer agent on nasopharing cancer is acetogenin, a polyketide compound that is abundant in Annona muricata L. leaves. This study has been done to examine polyketide isolation from Annona muricata  L. which has potency to induce apoptosis by p53 expresion on raji cell line. Methods: Cytotoxic effect was tested by MTT assay and expression of p53 was evaluated by immunihistochemistry on raji cell line while the identification of polyketide compound from Annona muricara L. leaves was evaluated by FTIR and UV-Vis. Result: Data analysis showed that polyketide isolate from Annona muricata L. leaves could enhance p53 pathway significantly on raji cell line.

35. Tuberculosis-Related to Knowledge, Adverse Drug Reactions, Clinical Outcome, Adherence in Tuberculosis Patients and Pharmacist Role, A Preliminary Survey for Pharmacist Intervention Model Development
Nanang Munif Yasin, Djoko Wahyono, Bambang Sigit Riyanto, Ika Puspita Sari
Tuberculosis (TB) is considered as a major problem in Indonesia. Many studies have been conducted in past in tuberculosis study, however study focus on role of pharmacist in TB is still limited. The objectives of this study were to explore and to observe baseline data in TB patients and the role of the pharmacist in TB management. A prospective, observational study was conducted in 21 primary health care and 4 hospitals in Yogyakarta. The data was collected through questionnaires, medical records, and interviewed with patients and pharmacists. Collecting the data were analysed descriptively. The analysis was carried out on 199 patients with inclusion criteria and it was found that 59,8% were male, 40,7% secondary educated, and 46,7% smokers. Tuberculosis-related to knowledge score was intermediate in 54,3% patients, especially in the side effect, TB resistant and TB transmission. Out of 199 patients 51,8% patients had an experience on adverse drug reactions. The most frequent ADR was gastrointestinal intolerance in 20,1% patients. The clinical outcome of the treatment included sputum culture conversion in 85,4 % patients, and gained weight in 78,9 % patients. There were 91,0 % patients had high adherence.  Although the role of the pharmacist has been established in terms of providing TB medicines and information when dispensing, but it could still be improved through a comprehensive intervention program that includes training, education, monitoring, adherence, and collaboration with other TB team.

36.Preparation and Characterization of Meloxicam-β-Cyclodextrin Inclusion Complex Prepared by Spray Dring Method
Fuad Soegibudiono Wiradjaja, Teuku Nanda Saifullah Sulaiman, Abdul Rohman
Development of meloxicam (MLX) oral dosage form has its own problem due to it’s poor water solubility and bitter taste. Several techniques have been used to improve the solubility and to mask the bitter taste of MLX, mainly by its complexation with cyclodextrins. The objective of the present work is to enhance the solubility of MLX and to prepare and characterize spray dried inclusion complex with β-cyclodextrin. Phase solubility study was done to analyze the influence of β-cyclodextrin on solubility of MLX. Inclusion complex of MLX- β-cyclodextrin was prepared in 1:2 molar ratio and conducted by spray drying method. Inclusion complex obtained was studied by organoleptic, Differential Scanning Calorimetry (DSC), and Fourier Transform Infrared Spectroscopy (FTIR). Phase solubility study shows that MLX solubility increase linearly with an increase in the concentration of  β-cyclodextrin. The addition of 3.48 mM β-cyclodextrin in phosphate buffer results in enhance of solubility of MLX about 25%. Furthermore, the addition of 31.75 mM β-cyclodextrin can enhance solubility of MLX up to 252%. Organoleptic analysis shows that inclusion complex generated a tasteless powder with an amoprhous and paler colour. DSC and FTIR analysis confirm the inclusion of MLX in the cyclodextrin cavity, even some functional groups of MLX are detected in the inclusion complex.

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