scholarly journals SIMPLE AND RAPID METHOD FOR THE SIMULTANEOUS ANALYSIS OF TAMOXIFEN, ENDOXIFEN, AND 4-HYDROXYTAMOXIFEN IN DRIED BLOOD SPOT USING LIQUID CHROMATOGRAPHY–TANDEM MASS SPECTROMETRY

2020 ◽  
pp. 112-120
Author(s):  
BAITHA PALANGGATAN MAGGADANI ◽  
YAHDIANA HARAHAP ◽  
HARMITA ◽  
SAMUEL J. HARYONO ◽  
TESANIKA RIBKA JOULIN SITORUS
Keyword(s):  
Formic Acid ◽  
Mobile Phase ◽  
Internal Standard ◽  
Cancer Recurrence ◽  
Dried Blood Spot ◽  
Active Metabolites ◽  
Drying Time ◽  
Liquid Chromatography Tandem Mass ◽  
Blood Spot ◽  
Spot Volume

Objective: Tamoxifen (TAM) is a hormonal therapy that is clinically proven to reduce breast cancer recurrence by blocking estrogen receptor, mainly through its active metabolites, 4-hydroxytamoxifen (4HT) and endoxifen (END), which have a higher affinity to ER than TAM itself. The objective of the present study was to develop and validate simple and rapid LC-MS/MS method for analysis TAM and its metabolites simultaneously in dried blood spot (DBS) sample for monitoring studies purposes. Methods: Optimization was done by evaluating several parameters that affect the efficiency of DBS preparation, such as blood spot volume, drying time and extraction method from the DBS paper. The effectiveness of chromatographic conditions was also optimized by varying flow rate, mobile phase combination and gradient. Clomiphene was used as the internal standard. Results: The result showed that preparation of 20 µl blood spot volume with 120 min of drying time and 25 min of extraction time using 1 ml methanol was the most efficient condition and also fulfilled recovery and matrix effect requirement according to FDA and EMA guidelines. The separation was performed on UPLC Class BEH C18 using formic acid 0.1%-formic acid 0.1% in acetonitrile (35:65) as the mobile phase in isocratic mode at 0.25 ml/min with a total analysis time of 4 min. Conclusion: This method has successfully fulfilled all validation requirements referring to EMA and FDA guidelines.

scholarly journals VALIDATION OF 6-MERCAPTOPURINE AND 6-THIOGUANINE IN DRIED BLOOD SPOT BY ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY

2018 ◽  
Vol 10 (1) ◽  
pp. 412
Author(s):  
Yahdiana Harahap ◽  
Cheputri Rahma Astrini ◽  
Herman Suryadi
Keyword(s):  
Liquid Chromatography ◽  
Formic Acid ◽  
High Performance ◽  
Gradient Elution ◽  
Ultra Performance Liquid Chromatography ◽  
Dried Blood Spot ◽  
European Medicines Agency ◽  
Tandem Mass ◽  
Liquid Chromatography Tandem Mass ◽  
Blood Spot

Objective: This study aimed to obtain an optimal and validated method of analyzing 6-mercaptopurine (6-MP) and 6-thioguanine (6-TG) simultaneouslyin dried blood spot samples using ultra high-performance liquid chromatography-tandem mass spectrometry.Method: Separation was performed with a 1.7-μm amide column, which had a mobile phase with a flow rate of 0.2 mL/min and comprised 0.2%formic acid in water, 0.1% formic acid in acetonitrile, and methanol with a gradient elution. Detection was performed using Waters Xevo TQD.Result: This method was linear with a range of 25–1000 ng/mL for 6-MP and 6-TG, with consecutive r values of ≥0.996 and ≥0.995, respectively. Theintra- and inter-day % difference value and coefficient of variation for the accuracy and precision were not more than 15% and 20%, respectively, ata concentration lower limit of quantitation.Conclusion: This method fulfilled the requirements of the European Medicines Agency guideline for validation.

scholarly journals Simultaneous Analysis of 6-Mercaptopurine, 6-Methylmercaptopurine, and 6-Thioguanosine-5’-monophosphate in Dried Blood Spot Using Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

2018 ◽  
Vol 18 (3) ◽  
pp. 544 ◽  
Author(s):  
Supandi Supandi ◽  
Yahdiana Harahap ◽  
Harmita Harmita ◽  
Rizka Andalusia
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Ultra Performance Liquid Chromatography ◽  
Dried Blood Spot ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Blood Spot

6-Mercaptopurine is a chemotherapeutic agent of the antimetabolite class. This study aims to analyze simultaneous validation of 6-mercaptopurine (6-MP), 6-methylmercaptopurine (6-MMP), and 6-thioguanosine-5’-monophosphate (6-TGMP) in dried blood spot (DBS) using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). An accurate volume of 60 μL blood was spotted onto DBS-CAMAG paper and then extracted using methanol 90% (v/v) containing an internal standard of 5-fluorouracil (5-FU). Separation was performed using a Waters Acquity UPLC BEH AMIDA column 1.7 μm (2.1 x 100 mm) with a mobile phase mixture of 0.2% (v/v) formic acid in water−0.1% (v/v) formic acid in acetonitrile-methanol with gradient elution and flow rate of 0.2 mL/min. Mass detection was done using Waters Xevo TQD with positive electrospray ionization (ESI) for 6-MP, 6-MMP, 6-TGMP and negative ESI for 5-FU, in multiple reaction monitoring mode. Detection rates of 6-MP, 6-MMP, 6-TGMP and 5-FU were m/z 153.09 > 119.09; 167.17 > 126.03; 380.16 > 168.00); 129.09 > 42.05, respectively. This method is linear across the range 25.5–1020 ng/mL for 6-MP, 6-MMP and 6-TGMP. This method is valid for the in vitro simultaneous analysis of 6-MP, 6-MMP and 6-TGMP in DBS, based on European Medicine Agency guidelines.

scholarly journals FACTORS AFFECTING LIQUID CHROMATOGRAPHY TANDEM-MASS SPECTROMETRY ANALYSIS OF 6-MERCAPTOPURINE AND 6-THIOGUANINE IN DRIED BLOOD SPOTS

2018 ◽  
Vol 10 (1) ◽  
pp. 200
Author(s):  
Yahdiana Harahap ◽  
Dimas Agus Putera Hardijanto ◽  
Delly Ramadon
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Formic Acid ◽  
Internal Standard ◽  
Standard Addition ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Blood Spot

Objective: This study aimed to determine the effects of the method of internal standard addition, spotting volume, paper type, and sample storagetemperature on 6-mercaptopurine, and 6-thioguanine on liquid chromatography tandem-mass spectrometry (LC-MS/MS) bioanalysis methods usingdried blood spot (DBS).Methods: Blood samples were spotted on CAMAG DBS paper and a Perkin Elmer 226 sample collection device (paper) and extracted into methanolcontaining 5-fluorouracil as an internal standard. The separation was performed on a water acquity ultra high-performance LC BEH Amide 1.7 μm(2.1 mm×100 mm) column with a mobile phase of 0.2% formic acid in water - 0.1% formic acid in acetonitrile methanol with gradient elution at aflow rate of 0.2 mL/min.Results: The step at which the internal standard was added (blood, spot on DBS card, or extraction solution) affected the chromatogram. Differencesin paper types and blood volumes significantly affected (p<0.05) the percent coefficient of variation, whereas differences in blood hematocritsignificantly affected the peak area ratio.Conclusion: The method of internal standard addition affected the chromatograms in this study. The best chromatogram was observed whenthe Internal Standard was added to the extracting solution. The card type also affected the analysis, so it is recommended to use the same card duringsample analysis.

scholarly journals Development and Validation of a Sensitive, Specific and Reproducible UPLC-MS/MS Method for the Quantification of OJT007, A Novel Anti-Leishmanial Agent: Application to a Pharmacokinetic Study

2021 ◽  
Vol 18 (9) ◽  
pp. 4624
Author(s):  
Maria Rincon Nigro ◽  
Jing Ma ◽  
Ololade Tosin Awosemo ◽  
Huan Xie ◽  
Omonike Arike Olaleye ◽  
...  
Keyword(s):  
Formic Acid ◽  
High Performance ◽  
Leishmania Major ◽  
Internal Standard ◽  
Gradient Elution ◽  
Rat Plasma ◽  
The Matrix ◽  
Liquid Chromatography Tandem Mass ◽  
Positive Mode ◽  
Acquity Uplc

OJT007 is a methionine aminopeptidase 1 (MetAP1) inhibitor with potent anti-proliferative effects against Leishmania Major. In order to study its pharmacokinetics as a part of the drug development process, a sensitive, specific, and reproducible ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated. Voriconazole was used as the internal standard to generate standard curves ranging from 5 to 1000 ng/mL. The separation was achieved using a UPLC system equipped with an Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm) with 0.1% formic acid in acetonitrile and 0.1% formic acid in water as the mobile phase under gradient elution at a flow rate of 0.4 mL/min. The mass analysis was performed with a 4000 QTRAP® mass spectrometer using multiple-ion reaction monitoring (MRM) in the positive mode, with the transition of m/z 325 → m/z 205 for OJT007 and m/z 350 → m/z 101 for voriconazole. The intra- and inter-day precision and accuracy were within ±15%. The mean extraction recovery and the matrix effect were 95.1% and 7.96%, respectively, suggesting no significant matrix interfering with the quantification of the drug in rat plasma. This study was successfully used for the pharmacokinetic evaluation of OJT007 using the rat as an animal model.

scholarly journals Clinical Validation of a Dried Blood Spot Assay for 8 Antihypertensive Drugs and 4 Active Metabolites

2020 ◽  
Vol 42 (3) ◽  
pp. 460-467
Author(s):  
Laura E. J. Peeters ◽  
Lida Feyz ◽  
Edon Hameli ◽  
Tom Zwart ◽  
Soma Bahmany ◽  
...  
Keyword(s):  
Antihypertensive Drugs ◽  
Dried Blood Spot ◽  
Clinical Validation ◽  
Active Metabolites ◽  
Blood Spot

scholarly journals The influence of the dried blood spot drying time on the recoveries of six immunosuppressants

2015 ◽  
Vol 1 (4) ◽  
pp. 116-122 ◽  
Author(s):  
Remco A. Koster ◽  
Rixt Botma ◽  
Ben Greijdanus ◽  
Donald R.A. Uges ◽  
Jos G.W. Kosterink ◽  
...  
Keyword(s):  
Dried Blood Spot ◽  
Drying Time ◽  
Blood Spot

Candidate reference method for determination of vitamin D from dried blood spot samples

2020 ◽  
Vol 58 (5) ◽  
pp. 817-827 ◽  
Author(s):  
Rosita Zakaria ◽  
Katrina J. Allen ◽  
Jennifer J. Koplin ◽  
Peter Roche ◽  
Ronda F. Greaves
Keyword(s):  
Vitamin D ◽  
Internal Standard ◽  
Reference Method ◽  
Population Level ◽  
Population Based ◽  
Dried Blood Spot ◽  
Coefficient Of Determination ◽  
Plasma Vitamin ◽  
Blood Spot

AbstractBackgroundThe current millennium has seen an explosion in vitamin D testing with the overarching aim of requests to clinically stratify patients as replete or deficient in vitamin D. At a population level, dried blood spot (DBS) sampling offers a less invasive and more practical application for assessment of vitamin D status. We have therefore aimed to develop a sensitive and robust DBS vitamin D method that is traceable to serum for use in population-based studies.MethodsBlood spots, calibrators and controls were prepared by punching a 3.2 mm DBS from filter paper and placed into a 96-well micro-plate. The DBS disk was eluted with a combination of water-methanol and internal standard (ISTD) solution followed by supported-liquid extraction and derivatisation. The extract was analysed by liquid-chromatography tandem-mass spectrometry in positive electrospray-ionisation mode with 732.5 > 673.4 and 738.4 > 679.4 m/z ion-transitions for derivatised vitamin D and the ISTD, respectively. Vitamin D results were made traceable to the National Institute of Standards and Technology reference material through the inclusion of Chromsystems vitamin D calibrators.Results25-Hydroxy-vitamin D3 and its related ISTD were detected at a retention time of 7 min. The seven-point calibration-curve consistently demonstrated a coefficient of determination of 0.99 with an experimentally determined reportable range of 0.5–376 nmol/L. Method validation studies using DBS samples demonstrated 12.9% between-assay imprecision at 45 nmol/L, 84% average recovery and high correlation with plasma vitamin D (correlation coefficient = 0.86).ConclusionsWe have successfully developed an analytical method for vitamin D quantitation from DBSs which will be applied to our population-based vitamin D research study. This approach improves traceability of DBS results and potentially could be used broadly for other DBS measurands that require comparison to serum/plasma for their interpretation.

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