Indonesian Journal of Pharmacy

Beta-Blocker in Heart Rate Control and Cardio Protection: The Role of ADRB1 Variants and HCN4 Regulation – A Systematic Review

Rasmaya Niruri, Zullies Ikawati, Agung Endro Nugroho, Habibie Arifianto — Wed, 25 Sep 2024 00:00:00 +0700

Elevated heart rate is linked to adverse cardiovascular outcomes. Sinoatrial (SA) nodes, hyperpolarization-activated cyclic nucleotide-gated-4 (HCN4) channels, and beta1-adrenergic receptor (ADRB1) are responsible for generating the heart rate. Beta-blockers have a cardioprotective effect on heart failure, including controlling heart rate. However, the responses to beta-blockers can vary among individuals. ADRB1 genetic variability may be contributed to the differential beta-blocker effect in heart failure. HCN4 also performs a crucial function in the pacemaker cells of the heart. Exploring the effect of beta-blockers in pacemaker cells is expanding the view of their role and their therapeutic response in heart failure. The objectives of this study were to identify ADRB1 genetic variants affecting heart rate response in heart failure subjects with beta-blocker treatment and to explore the effect of beta-blockers on HCN4 channels and SA nodes. A systematic review was performed using three databases. Eight of 668 manuscripts were selected. The systematic review found that ADRB1 genetic variants (A145G (Ser49Gly) and C1165G (Arg389Gly)) can affect heart rate response in beta-blocker-treated heart failure. The study also found that the percentage of patients with the Ser49Ser-Gly389X haplotype achieved a heart rate target was higher than other haplotypes. Individuals with the Arg389Arg genotype necessitated a markedly increased amount of beta-blocker dose to reach the identical heart rate target compared to those with the Gly389X gene variation. In addition, the review found that carvedilol, a beta-blocker derivative, demonstrated beneficial effects in inhibiting HCN-gated channels. Bisoprolol and carvedilol improved channel regulation in the SA Node by reversing the downregulation of HCN4 and sodium channels. In general, this systematic review provides important insights into beta-blockers in treating heart failure, specifically concerning the genetic variability of ADRB1 and the beta-blockers effect on the SA node and HCN4 channels.

Clinical Decision Support Systems to Identify Drug-Related Problems in Diabetes Mellitus Patients: A Systematic Review

Niken Larasati, Satibi Satibi, Susi Ari Kristina, Lutfan Lazuardi — Wed, 25 Sep 2024 00:00:00 +0700

Clinical Decision Support Systems (CDSSs) has been developed for utilization to provide rational therapy to Diabetes Mellitus (DM) patients including preventing Drug-related Problems (DRPs) such as the effect of drug treatment being not optimal, untreated indications and symptoms, unnecessary therapy, and adverse drug reactions. This study aims to summarize the available evidence on the intervention of CDSSs, key outputs, and impact of the user in DM patients. This study was a systematic review using PubMed, Scopus database, and by manually searching the bibliographies of articles that have been found. We included studies reporting on evaluated CDSSs that had been implemented in medication prescription, reducing medication errors, adverse drug events, drug-allergy checking, drug dosing support, and so on. A total of 8 studies were selected among 855 studies. CDSSs are used in hospitals and primary care settings to identify potential drug interactions, correct therapy regimens, monitor therapy, blood glucose documentation, ensure patients receive medication according to the guideline, provide nutritional advice, and schedule physical activity. The usage of CDSSs improves blood glucose levels, detects possible drug interactions, reduces face-to-face consultations, improves documentation, assists in identifying dose, and promotes prescribing in line with the guideline. The use of CDSSs can help to reduce the risk of errors in management therapy.

Phytochemical, Nutraceutical and Pharmacological Aspects of the Philippine native Acalypha angatensis Blanco, Fl. Filip.

Wed, 25 Sep 2024 00:00:00 +0700

The genus Acalypha has valuable species that have significant pharmaceutical applications. This study investigated the phytochemical composition and thin layer chromatography (TLC) profile, as well as the antioxidant and antibacterial activities of the ethanolic extract of Acalypha pulogensis Sagun & G.A. Levin, an endemic species in the Philippines. Qualitative phytochemical screening revealed the presence of tannins, phenolics, flavonoids, saponins, steroids, and triterpenoids. Nutrient analysis showed that potassium and calcium content were at 1499 mg/100 g and 1380 mg/100 g, respectively. The determination of the total phenolic content (TPC) showed that this species has an average of 282.24 ± 3.56 mg GAE/g and a total flavonoid content (TFC) of 36.24 ± 3.77 mg QE/g of dried extract. These values were found to be higher than those of other Acalypha species. Antioxidant activity was measured using DPPH antioxidant assay, and the extract showed a concentration-dependent radical scavenging effect with an IC₅₀ of 34.02 µg/mL. Antibacterial activity was evaluated using various concentrations against drug-resistant Gram-positive and Gram-negative bacteria using a resazurin-based assay. The ethanolic extract exhibited substantial antibacterial activity against Enterococcus faecium (NCTC 12204) and MRSA (ATCC 33592), with a minimum inhibitory concentration (MIC) of 4 mg/mL. Higher concentrations (>20 mg/mL) were needed for inhibition of the Gram-negative strains (Klebsiella pneumoniae and Enterobacter cloacae). These results suggest that A. pulogensis has good antioxidant and antibacterial activities that support its use in traditional medicine.

Isolation of Nutmeg Essential Oil (Myristica fragrans houtt) From Aceh Indonesia and Their Antioxidant and Antibacterial Activities

Khairan Khairan, Meutia Faradilla, Binawati Ginting — Wed, 25 Sep 2024 00:00:00 +0700

This study aims to isolate the nutmeg essential oil (Myristica fragrans Houtt) from Aceh Indonesia and evaluate their biological activity. The antioxidant activity of the nutmeg essential oils (Myristica fragrans Houtt) was determined by DPPH (1,1-difenil-2-pikrihidrazil) assay. The DPPH assay showed that arillus essential oil (AMFH) has the highest antioxidant activity with the IC₅₀ values was 216,695 ppm. The chemical composition of the AMFH was perfomed by Gas Chromatography-Mass Spectroscopy (GC-MS), and the results showed that AMFH contains Terpinen-4-ol (T-4-ol), α-Terpineol (α-T), Safrole (Saf) and Myristicin (Myr) with the percentage area of 11.20%; 1.83%; 5.10%; and 27.80% respectively. The AMFH was isolated by column chromatography and four fractions were obtained, namely AMFH1; AMFH2; AMFH3; and AMFH4 fractions. The DPPH assay showed that the AMFH3 fraction showed excellent antioxidant activity with the IC₅₀ values of 59.329 ppm. The AMFH3 fraction was than reisolated by using by column chromatography to obtain four subfractions namely AMFH3A; AMFH3B; AMFH3C; and AMFH3D. The DPPH assay showed that the subfraction of AMFH3A has the highest antioxidant activity with the IC₅₀ values of 98.993 ppm. The antibacterial activity of AMFH3A subfractions was evaluated by Kirby Bauer-Disc diffusion method, and the results showed that AMFH3A subfraction showed slightly higher activity against Staphylococcus epidermidis and Staphylococcus aureus with the diameter inhibition of 14.28 mm and 12.26 mm respectively. The chemical composition of the AMFH3A subfraction was performed by GC-MS, and the results showed that AMFH3A subfraction contains Terpinen-4-ol (T-4-ol), α-Terpineol (α-T), Safrole (Saf) and Myristicin (Myr) with the percentage area of 14.37%; 2.16%; 7.36%; and 54.77% respectively.

Gut Microbiota Dynamics and Phenotypic Changes Induced by Tetracycline in Drosophila melanogaster

Wed, 25 Sep 2024 00:00:00 +0700

Gut microbiota plays a crucial role in both physiological and pathological processes in humans and animals. Antibiotics, designed to combat bacterial infections, can induce alterations in the composition and abundance of the gut microbiota over prolonged exposure. This study addresses the limited understanding of the connections between gut microbiota and phenotypic profiles of metazoan species. We investigated the impact of early-life exposure to tetracycline in wild-type D. melanogaster, which were fed a standard diet, comparing them to a control group not exposed to tetracycline. The primary objective was to examine the consequences of early-life tetracycline exposure on gut microbiota and its implications for phenotypic profiles, including survival, locomotor activity, and reproduction in adult flies. Results revealed a significant reduction in lactic acid bacteria in adult flies exposed to tetracycline. However, tetracycline exhibited no interference with fly development, allowing them to maintain a normal lifespan. In adult flies, tetracycline significantly decreased the lifespan on day 35 at a concentration of 1000 µg/mL and reduced locomotion on day 27 at concentrations of 10 µg/mL and 1000 µg/mL. Remarkably, tetracycline did not impact the reproductive capabilities of the flies. This study demonstrates that while tetracycline led to a decline in lactic acid bacteria, locomotion, and lifespan in adult flies, it did not disrupt their development or reproductive processes.

PyPLIF HIPPOS-aided Construction and Retrospective Validation of Structure-Based Virtual Screening Protocol Targeting VEGFR2

Wed, 25 Sep 2024 00:00:00 +0700

Recently, the discovery of small molecules as inhibitors for vascular endothelial growth factor receptor 2 (VEGFR2) is of timely interest, especially in the area of ocular neovascular diseases. On the other hand, PyPLIF HIPPOS in combination with machine learning techniques has been reported to increase the prediction quality of structure-based virtual screening (SBVS) protocols. The original version of PyPLIF has served in the development of an SBVS protocol that successfully identified novel chalcone derivatives and short peptides as potent inhibitors for acetylcholinesterase. In this short communication, construction and retrospective validation of an SBVS protocol employing PyPLIF HIPPOS targeting VEGFR2 are presented to make it publicly available. The retrospective validation employed 409 active compounds and 24,950 decoys from the enhanced version of the directory of useful decoys. All compounds were docked independently 3 times using AutoDock Vina followed by the identification of the protein-ligand interaction fingerprints (PLIF) employing PyPLIF HIPPOS. The derived ensemble PLIF descriptors were then used in the decision tree construction using a machine-learning technique called recursive partitioning and regression trees. The best decision was then incorporated in the SBVS protocol. The F-measure and enrichment factor values of the SBVS protocol were 0.387 and 76.879, respectively. Hence, the SBVS protocol is readily available to screen small molecules or short peptides.

Comparative Cellular and In Vivo Anti-cancer studies of Doxorubicin Liposomes Prepared with Different Types of Phospholipids

Andang Miatmoko, Devy Maulidya Cahyani, Kumi Kawano, Yoshiyuki Hattori — Wed, 25 Sep 2024 00:00:00 +0700

The selection of lipid components of membrane bilayer determines the rigidity of liposomes affecting drug efficacy, especially for cancer drug delivery. The present study evaluated liposomes with different rigidity for delivering doxorubicin (DOX). In this work, liposomes composed of rigid lipid, hydrogenated soybean phosphatidylcholine (HSPC), were totally or partially substituted with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). The liposomes are composed of phosphatidylcholine (HSPC, POPC), with and without combination with DOPE, cholesterol, and DSPE-mPEG₂₀₀₀ with a molar ratio of 57: 38: 5, respectively. Liposomes were prepared using a thin layer hydration method. Then, in vitro cytotoxicity and in vivo antitumor activity of these liposomes were evaluated. Substitution of HSPC with POPC resulted in similar cytotoxicities profiles similar to the DOX solution on C26 colon cancer cells and LLC cells. The DOPE addition to DOX liposomes reduced the antitumor activity. In conclusion, the lipid substitution of HSPC with POPC or DOPE reduced liposome rigidity; however, it lowered the in vivo antitumor activity.

Zinc Sulfate and α-tocopherol Supplementation Enhance Reproductive Performance in Male Albino Rats (Rattus norvegicus) With Lead Acetate Toxicity

Wed, 25 Sep 2024 00:00:00 +0700

Metal toxicity from lead affects reproductive organ function by activating reactive oxygen species processes. This study aims to see how α-tocopherol and zinc sulfate (ZnSO4) affect gonads, liver, follicle-stimulating hormone, luteinizing hormone, spermatogenesis (the amount of spermatogonia, spermatocytes, and spermatids), and Leydig cells in male albino rats (Rattus norvegicus) exposed to lead acetate Pb(CH₃COO)₂. The samples used were 25 male Wistar rats aged 4 months, separated into five groups. For 30 days, all treatment groups were exposed to Pb(CH₃COO)₂ at a level of 50-mg/kg body weight (BW). The T1 group was given a dosage of 100-mg/kg BW of α-tocopherol. The ZnSO4 was given to the T2 group at a dose of 0.54-mg/kg BW. Meanwhile, the T3 group was given a mixture of ZnSO4 at 0.54-mg/kg BW and α-tocopherol at 100-mg/kg BW orally. ELISA test was carried out to determine the level of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in a blood plasma sample of 100 μl / 1 mg. Histopathological observations made on the liver included counting damaged cells and seminiferous tubules that included counting the amount of spermatogonia, spermatocytes, spermatids, Sertoli cells, and Leydig cells. Using SPSS 20 software, the collected data were analyzed using analysis of variance, followed by Duncan’s test with a 95% simultaneous confidence level. The highest average levels of FSH and LH in the T3 group were 3.6162 mIU/mL and 14.9658 mIU/mL. The finding showed that Pb(CH₃COO)₂ caused disruptions in the spermatogenesis and Leydig cell processes. Exogenous antioxidants in combination with ZnSO4 and α-tocopherol had significant effect on enhancing reproductive performance in animals exposed to Pb(CH₃COO)₂.

Attenuation of TNF-α and Iron Levels in Renal Hemosiderosis by Phaleria macrocarpa (Scheff.) Boerl Extract in a Rat Iron Overload Model

Wed, 25 Sep 2024 00:00:00 +0700

Regular blood transfusions are typically used to treat the genetic anemia known as thalassemia, which can lead to an increase in the body's total iron levels. The condition of excess iron can be toxic to the body due to the formation of free radicals that are harmful and can damage cells and tissues. So the availability of free iron will form the basis of iron toxicity because it accelerates the Fenton reaction to produce an increase in the amount of ROS that cannot be suppressed so that saturation of the antioxidant system can occur. Excess iron has been known to be a risk factor for organ dysfunction and damage that results in various organ diseases such as liver, heart and kidney, diabetes mellitus, and neurodegenerative diseases. Mangiferin is an active compound has been shown to act as an iron chelating agent by forming complexes with iron. The complex formed can reduce iron accumulation in thalassemia patients who receive blood transfusions on a regular basis. Mahkota Dewa (Phaleria macrocarpa) is a well-known medicinal plant native to Papua, Indonesia, and it also includes the active ingredient mangiferin. This study aims to determine the effectiveness of the ethanolic extract of Phaleria macrocarpa fruit as an iron chelating agent observed in the kidneys in a rat model of excess iron

Physico-Chemical and Structural Characterization of Mixed Natural Polymer Hydrogels Under Simulated Gastrointestinal Fluid

Wed, 25 Sep 2024 00:00:00 +0700

The gastrointestinal track has a varied pH, from an acidic pH in the stomach to a slightly basic pH in the intestine. The pH difference creates problems for the delivery of drugs and nutrients, as the active compounds may not survive the changing pH. Incorporating active ingredients into hydrogels can protect the compounds from degradation. Natural polymer hydrogel is preferable because of its safety and compatibility. However, a suitable formula should be optimized to facilitate suitable delivery in the gastrointestinal track. In this study, we produced hydrogels with 10 different formulas of mixed natural polymers: CMC (C), alginate (A), chitosan (X), and/or guar gum (G). The resulting hydrogels were characterized using swelling performance tests at pH 2 and 6.8, Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). Hydrogels with the formula of C50A25X25, C25A50X25, C25A50G25, C25A25G50, C25A50X12.5G12.5, and C25A25X25G25 are not digested at pH 2, but those hydrogels are digested at pH 6.8. The FTIR spectra of the hydrogels show functional groups of O–H, C=O, C–O, and C–H. Meanwhile, the SEM results show cracking phenomena on the surface of the hydrogels C50A25X12.5G12.5, C25A50X12.5G12.5, and C25A25X25G25. This study offers a general guide for the development of a natural polymer hydrogel as a suitable nutrient vehicle in the gastrointestinal track. Formulation of each type of nutrient should be optimized for optimum delivery to the designated part of the gastrointestinal track.

Optimizing the Formula of Polymeric-Based Aripiprazole Nanosuspension Using Response Surface Methodology for Intranasal Drug Delivery

Nabila Fatin Aisiah, Silvia Surini — Wed, 25 Sep 2024 00:00:00 +0700

This study aimed to optimize the formula of aripiprazole nanosuspension for intranasal drug delivery. Response Surface Methodology (RSM) was employed to determine the influence of independent variables, including drug concentration, polymer concentration, and the ratio of polymer combination, on the nanosuspension characteristics. The parameters under investigation were particle size (d mean), polydispersity index, and drug content. Fifteen formulas were prepared using the high-shear homogenization–ultrasonication method, and the Design Expert software was applied for optimum formula determination. The result showed significant effects of the independent variables on the nanosuspension characteristics, with particle sizes ranging from 143.6 – 334.6 nm, PDI values of 0.302 – 0.649, and drug content was 98.7 – 102.1 %. The predicted optimum formula had a drug concentration of 28 mg/mL in the organic solvent, polymer concentration of 1.5% (w/v), and HPMC to PVP ratio of 1.4 with desirability of 0.94. Additionally, it exhibited desirable characteristics, such as a particle size of 171.2  11.4 nm, a PDI value of 0.317  0.02, and a high drug content of 100.04  0.65%. In conclusion, the presented methodology appeared to be perfect for the optimization of the aripiprazole nanosuspension formula to ensure suitability for nose-to-brain drug delivery.

Simultaneous Quantification of Ramipril and Ramiprilat in Drug Formulations: A Clinical LC-MS/MS Study

Wed, 25 Sep 2024 00:00:00 +0700

Ramipril is a drug controlling hypertension. Two drugs were studied in 36 healthy adults. The drugs are the test (Atb^®) and the reference (Tritace^®). This concept is crucial in the pharmaceutical industry during the development process. The maximum concentration (C_max) of Ramipril and its metabolite (Ramiprilat) were determined throughout drug administration in healthy subjects. All analysis was performed using LC-MS/MS. The method was validated over a range of 0.200 – 25.000 ng/mL in which linearity (R²) was 0.9983, accuracy was 99.9%, precision (CV) was less than 20%, and the Lower Limit of Quantitation was 0.200 ng/mL. Stability and other parameters were discussed. The time needed to reach this concentration (T_max) and the time-drug concentration area under the curve (AUC) were most agreed upon. The two drugs were proven to be interchangeable. Both have similar pharmacokinetics and bioequivalence profiles. Consequently, the drugs are bioequivalent and considered alternatives to one another pharmaceutically. For example, 90% confidence intervals (C.I.) for the in-tra-individual ratios (test/reference) for pharmacokinetic parameters were C_max, and AUC_0-t for total exposure C.I. Log – transformed Values were 80-125 %.

Formulation of Lotion with Ethanol Extract of Curcuma manga Val. as UV B Sunscreen and Activity Test using UV-Vis Spectrophotometry

Nindya Kusumorini, Abdul Karim Zulkarnain — Wed, 25 Sep 2024 00:00:00 +0700

Temu mango (Curcuma manga Val) is a rhizome plant native to Indonesia that contains curcuminoid and flavonoid antioxidant compounds. These compounds are thought to have activity as UV light protectors, especially UV B. This study uses a qualitative test of the compound content in temu mango followed by formulation and activity testing of water in oil (w/o) sunscreen lotion of temu mango in vitro. Optimization of w/o lotion formulation uses Design Expert^® (DE) software version 9.0.4.1 with the Simplex Lattice Design method. The DE software obtained 13 runs of w/o lotion formula and continued with physical properties evaluation to determine the optimum formula. The optimum formula of lotion w/o predicted by DE software was tested for physical stability for four weeks, SPF test, percent pigmentation, and percent erythema using UV-Vis spectrophotometry. The results of the study show that temu mango extract was found to provide an SPF value at a concentration of 0.3 mg/mL with an SPF value of 16.62. The results of the optimum formula of lotion w/o were 7%glycerin, 7% cera alba, 5% cetyl alcohol with a value of stickiness of 0.70 ± 0.03 seconds, spreadability of 13.20 ± 0.83 cm², and viscosity of 95.19 ± 3.06 dPa.s. Referring to the storage results, the optimum formula of w/o lotion from temu mango extract was stable during one month of storage. The w/o lotion in the optimum formula produced an SPF value of 15.06 ± 0.39, percent erythema of 10.95%, and percent pigmentation of 10.41%.

The Role of Growth Differentiation Factor-15 in the Diagnosis of Patients with Chronic Angina Undergoing Coronary Angiography

Aseel Butti, Wassan Abdul Kareem Abbas, Ahmed Yousif Hasan — Wed, 25 Sep 2024 00:00:00 +0700

Coronary artery disease (CAD) is considered the most prevalent leading cause of myocardial ischemia and mortality worldwide and can lead to angina pectoris and myocardial infarction (MI). Growth Differentiation Factor-15 (GDF-15) is usually measured together with other biomarkers to predict all-cause mortality. During stressful conditions associated with tissue injury and inflammation like myocardial ischemia, it is released into circulation and detected at high concentration in foam cells of the atherosclerotic plaque. Its circulating levels are associated with chronic diseases rather than acute diseases mainly cardiovascular diseases (CVDs) like CAD and chronic heart failure (CHF). The aim of the current study was to detect the role of GDF-15 as a new biomarker for the diagnosis of stable angina patients with normal left ventricular ejection fraction who were presented as having stable chest pain, so that to avoid endangering them to the invasive coronary angiography. Fasting venous blood samples were taken from 90 participants who were presented as having chest pain. They were all subjected to echocardiography, electrocardiography and coronary angiography. The left ventricular ejection fraction was calculated using the biplane M mode method. GDF-15 serum level was measured by using Human GDF-15 Sandwich ELISA Kit following the manufacturer’s instructions.The current study detected a high significant difference in the serum levels of GDF-15 and serum creatinine between the patients and the control group (P≤0.01). On the other hand, there were significant differences in the serum levels of triglycerides and VLDL-C between both study groups (P≤0.05). ROC curve analysis showed that GDF-15 had AUC= 1.00, the best cut off= 254.16 with sensitivity and specificity of 100%. So, it can be concluded that GDF-15 could be used as a biomarker for the diagnosis of stable angina patients with obstructive atherosclerotic plaque and normal LV ejection fraction.

Validation of An HPLC Method for the Determination of Some Β-Lactams Antibiotics Using A Green Analytical Technique

Thoai Dang Nguyen, Anh Tran Thi Phuong Phuong, Ky Truong Quoc — Wed, 25 Sep 2024 00:00:00 +0700

Green analytical chemistry (GAC) primary goal is to develop environmental and health-friendly analytical methods by reduction or replacement of hazardous substances from analytical procedures. In this research, the application of GAC to quantify five β-lactams antibiotics (cefuroxime sodium, cefaclor, ertapenem sodium, ampicillin sodium and sulbactam sodium) by high-performance liquid chromatography (HPLC) using photodiode arrays (PDA) detector was developed. To achieve this, ethanol was used as a green replacement for methanol and acetonitrile, common solvents in chromatographic procedures. Several chromatographic conditions were investigated to accomplish the optimal conditions. The method validation followed the ICH Q2 (R1) guideline, and the developed method was successfully applied for quantifying some pharmaceutical products.

Analytical Method Validation and Simultaneous Determination of Ketotifen Fumarate and Cyproheptadine Hydrochloride in Divided Powder by RP-HPLC Method

Melynia Sintha Dewi, Florentinus Dika Octa Riswanto, Sri Hartati Yuliani — Wed, 25 Sep 2024 00:00:00 +0700

Ketotifen fumarate and cyproheptadine HCl are the most popular prescription for children at “X” hospital in Yogyakarta. The interest for studying this divided powder was related to the stability test towards human health. Hence, it was important to developed a validated analytical method to quantify content of active drug from sample. In this study, a reversed phase HPLC (RP-HPLC) method was validated and applied for determining content of ketotifen fumarate and cyproheptadine hydrochloride in divided powder. The developed RP-HPLC method was met the requirement of selectivity and sensitivity. Calibration curve formulas of ketotifen fumarate and cyproheptadine hydrochloride were y = 27714x-79111 (r=0.9945) and y = 26324x+72581 (r=0.9935) in the range of 10.9-54.5 and 10.6-53.0 µg/mL, respectively. Accuracy and intermediate precision were assessed on both standard solution and samples spiked with standard solution of ketotifen fumarate and cyproheptadine hydrochloride. It was found that both standard solution and the spiked samples solution were met the requirement of recovery of 80-110% and RSD of less than 7.3% for intraday and interday analysis. Content of ketotifen fumarate and cyproheptadine hydrochloride in sample (n=7) were 16.574±0.098 and 30.640±0.035 µg/mL, respectively. It can be concluded that the RP-HPLC method was successfully validated and applied in the determination of ketotifen fumarate and cyproheptadine hydrochloride in divided powder.