Yahdiana Harahap
Departemen Farmasi FMIPA, Universitas Indonesia

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Simultaneous Analytical Method Of 6-Mercaptopurine and 6-Methylmercaptopurine In-vitro Study With Bio-Sampling Venipuncture and Dried Blood Spot Yahdiana Harahap
Journal of Global Pharma Technology Volume 09 Issue 05
Publisher : Journal of Global Pharma Technology

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Abstract

Objective: 6-Mercaptopurine (6-MP) is a cancer chemotherapeutic agent. Metabolic pathway by thiopurine S-methyltransferase (TPMT) to become 6-methylmercaptopurine (6-MMP).Bio-sampling is required to obtainbiological sample, using two technique; invasive (venipuncture) and minimum invasive(dried blood spot). This study aims to obtain an optimization and validation analysis 6-MP and 6-MMP in vitro study with bio-sampling venipuncture and dried blood spot (DBS). Method: Plasma from venipuncture method extraction was done using dichloromethane. Separation was performed using Waters HPLC, C18 SunfireTM column (5μm, 250 x 4.6 mm), with gradient elution, flow rate 1 mL/min and detected at UV-PDA wavelength of 303 nm. Bio-sampling dried blood spot with DBS CAMAG® paper diameter of 8 mm and extracted with acetonitrile-methanol (1:3). Separation was performed with Waters LC-MS/MS UPLC C18 column (1.7 μm, 2.1 x 100 mm) with gradient elution, flow rate 0.2 mL/min. 5-fluorouracil (5-FU) was used as internal standard.Result: The method venipuncture was linear at concentration range of 2-200 ng/mL for 6-MP and 20-2000 ng/mL for 6-MMP. The method dried blood spot using Waters Xevo TQD for mass detection with positive electrospray ionization (ESI) for 6-MP, 6-MMP and negative ESI for 5-FU in Multiple Reaction Monitoring mode. Linear with the range 26-1000 ng/mL for 6-MP and 13-500 ng/mL for 6-MMP.Conclusion: The developed method is valid for 6-MP and 6-MMP simultaneously in vitro from venipuncture using HPLC and from dried blood spot using LC-MS/MS and showed good selectivity, linearity, accuracy and precision, matrix effect and stability.Keywords: 6-mercaptopurine, 6-methylmercaptopurine, Venipuncture, Dried blood spot.
Effect of Various Anticoagulants on Ethinyl Estradiol and Levonorgestrel Analysis in Human Plasma in Vitro by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry Yahdiana Harahap
Journal of Global Pharma Technology Volume 10 Issue 12.
Publisher : Journal of Global Pharma Technology

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Objective: This research objective is to evaluate the different types of anticoagulant to ethinyl estradiol and levonorgestrel analysis in plasma using ultra performance liquid chromatography tandem mass spectrometry. Method: Chromatography condition was obtained with Acquity® UPLC BEH C18 column (1.7 µm; 50 x 2.1 mm); mobile phase consisting 0.1% formic acid in water – acetonitrile under gradient elution ; flow rate of 0.3 mL/minute; column temperature of 40ºC; injection volume of 10.0 µL; 5-minute analysis time and prednisone as internal standard. Sample preparation used protein precipitation followed by liquid-liquid extraction. Results: There was a linear result ranging from 5-500 pg/mL concentration of ethinyl estradiol and 100-10,000 pg/ml concentration of levonorgestrel. There was no significant difference for stability and recovery of ethinyl estradiol and levonorgestrel in citrate, heparin, and EDTA plasma (p > 0.05; ANOVA). However, significant difference for peak area ratio (p < 0.05; Kruskal Wallis), between citrate, EDTA, and heparin plasma was observed. Conclusion: Citrate and heparin plasma analysis had better result than EDTA plasma analysis.Keywords: Ethinyl estradiol, Levonorgestrel, Prednisone, EDTA, Heparin, Citrate.
Determination of Valproic Acid without Derivatization in Human Plasma using High Performance Liquid Chromatography-Photodiode Array Yahdiana Harahap
Journal of Global Pharma Technology .
Publisher : Journal of Global Pharma Technology

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Abstract

Objective: To develop and validate a simple and sensitive HPLC method without derivatization for determination of valproic acid in human plasma. Methods: Nonanoic acid as internal standard was added to 500 mL of plasma sample prior to liquid-liquid extraction using n-hexane and 0.5% triethylamine. Chromatographic separation was achieved on C-8 Symmetry® column (5µm; 150 x 3.9mm) in isocratic mode at 45°C. The mobile phase was 40 mM sodium dihydrogen phosphate pH 3.5 – acetonitrile (56:44 %v/v) with flow rate of 1.00 mL/min and was detected at 210 nm. Method validation referred to EMA Guideline 2011 for bioanalytical method validation in term of parameters lower limit of quantification (LLOQ), selectivity, calibration curve and linearity, carry over, recovery and stability. The valid method was applied in pharmacokinetic study of one healthy subject after administration of 500mg extended release caplet of valproic acid. The blood was collected as much as 7mL for twelve spots, which are predose, 1, 2, 3, 4, 5, 8, 10, 24, 36, 48, and 72 hours after drug administration. Results: The calibration curve valproic acid was linear over the concentration range of 2.0 – 200.0 µg/mL (r = 0.9992). Within-run and between-run precision and accuracy were studied at four concentrations and RSDs were less than 1.8 % and 5.4 %, while the bias (accuracy values) were less than 17.9 % and 13.1 %, respectively. Conclusion: The developed method provides sensitivity, linearity, precision, accuracy and is suitable for analysis of valproic acid in plasma samples for pharmacokinetic studies.Keywords: Valproic acid, Nonanoic acid, HPLC, Validation, Liquid-liquid extraction.
Analysis of 4-Hydroxy-N-Desmethyltamoxifen and Tamoxifen in Dried Blood Spot of Breast Cancer Patients By Liquid Chromatography – Tandem Mass Spectrometry Yahdiana Harahap
Journal of Global Pharma Technology Volume 10 Issue 05: (2018) May2018
Publisher : Journal of Global Pharma Technology

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Abstract

Tamoxifen is the first line hormonal therapy for breast cancer patients as their adjuvant therapy. The antiestrogen effect of tamoxifen is highly determined by its active metabolite, endoxifen. A simultaneous quantification method of tamoxifen and endoxifen in dried blood spot (DBS) using LC-MS/MS had been fully validated in this study. Extraction the analyte and metabolite from DBS card were conducted using methanol-acetonitrile (50:50). The separation was performed on UPLC Class BEH C18 column using 0.2% formic acid - acetonitrile as the mobile phase in gradient elution mode at 0.2ml/min. The detection of the mass was performed on Waters Xevo TQD using positive electrospray ionization for tamoxifen, endoxifen, and clomiphene as the internal standard with m/z value: 372.22>72.22; 374.29>58.2; 406.28>100.17, respectively. This method was linear in the range concentration of 5-200 ng/ml for tamoxifen and 1-40 ng/ml for endoxifen with r value 0.99. The method was applied to 40 breast cancer patients, where the results lied between 40.28 and 194.10 ng/ml for tamoxifen, meanwhile for endoxifen was 1.25 and 18.02 ng/ml. It showed that there were 4 patients received less effective tamoxifen therapy based on the endoxifen threshold in DBS sample, which was 3.3 ng/ml. This method has prospect future to optimize tamoxifen therapy by measuring tamoxifen and endoxifen concentration. Keywords: Breast cancer; Dried blood spot; Endoxifen; Tamoxifen; Clomiphene; Analysis;  LC-MS/MS; Validation.
An Integrated Risk Analysis Approach in Military Hospitals: Implications for Public Health Preparedness and Resilience Harefa, Faonaso; Yahdiana Harahap; Dian Andriani Ratna Dewi; R.M. Tjahya Nurrobi; Sutanto Sutanto; Cecilia F. Harsono
Media Publikasi Promosi Kesehatan Indonesia (MPPKI) Vol. 9 No. 1: JANUARY 2026 - Media Publikasi Promosi Kesehatan Indonesia
Publisher : Fakultas Kesehatan Masyarakat, Universitas Muhammadiyah Palu

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.56338/mppki.v9i1.8697

Abstract

Introduction: Military hospitals perform a dual function by providing healthcare services for soldiers and their families while also supporting public health needs during crises. This dual role generates complex hazards spanning biological, chemical, physical, and psychological dimensions, thereby requiring a comprehensive risk analysis framework. The objective of this study is to develop an integrated risk analysis approach comprising risk assessment, risk management, and risk communicationto strengthen occupational safety in military hospitals, with broader relevance for public health and global health security. Methods: A mixed methods design was applied. Data were collected through direct observation and in-depth interviews with healthcare personnel, complemented by a structured survey using standardized questionnaires. Qualitative analysis was conducted using NVivo 12 and quantitative analysis using SEM PLS-4. The study involved 100 respondents comprising medical personnel, health workers, and staff at Rumah Sakit Pusat Pertahanan Negara (RSPPN) and Pusat Kesehatan TNI, selected through random sampling. Results: Qualitative findings derived from NVivo 12 analysis revealed a multidimensional hazard spectrum characterized by weak cross sectoral coordination, limited personnel capacity, and insufficient integration among risk analysis components. Quantitative analysis using SEM PLS-4 further confirmed that Integrated Risk Analysis has a positive and statistically significant effect on Public Health Preparedness and Community Resilience (T-statistic = 11.046 > 1.96; p-value < 0.05); and F-square (0.18- efect moderat). Conclusion: This study concludes that Integrated Risk Analysis exerts a significant influence on public health preparedness and community resilience in military hospitals. The findings underscore the necessity of strengthening management, integration, and cross sectoral communication. Nonetheless, the contextual limitations regarding research setting and sample size suggest the need for future studies with broader scope and institutional diversity to reinforce the generalizability of the findings.