scholarly journals Use of Dried Blood Spot Specimens to Monitor Patients with Inherited Metabolic Disorders

2020 ◽  
Vol 6 (2) ◽  
pp. 26 ◽  
Author(s):  
Stuart J. Moat ◽  
Roanna S. George ◽  
Rachel S. Carling
Keyword(s):  
Metabolic Disorders ◽  
Ultra Performance Liquid Chromatography ◽  
Dried Blood Spot ◽  
Specificity And Sensitivity ◽  
Low Concentrations ◽  
Inherited Metabolic Disorders ◽  
The 1960S ◽  
Analytical Technology ◽  
Test Result ◽  
Blood Spot

Monitoring of patients with inherited metabolic disorders (IMDs) using dried blood spot (DBS) specimens has been routinely used since the inception of newborn screening (NBS) for phenylketonuria in the 1960s. The introduction of flow injection analysis tandem mass spectrometry (FIA–MS/MS) in the 1990s facilitated the expansion of NBS for IMDs. This has led to increased identification of patients who require biochemical monitoring. Monitoring of IMD patients using DBS specimens is widely favoured due to the convenience of collecting blood from a finger prick onto filter paper devices in the patient’s home, which can then be mailed directly to the laboratory. Ideally, analytical methodologies with a short analysis time and high sample throughput are required to enable results to be communicated to patients in a timely manner, allowing prompt therapy adjustment. The development of ultra-performance liquid chromatography (UPLC–MS/MS), means that metabolic laboratories now have the capability to routinely analyse DBS specimens with superior specificity and sensitivity. This advancement in analytical technology has led to the development of numerous assays to detect analytes at low concentrations (pmol/L) in DBS specimens that can be used to monitor IMD patients. In this review, we discuss the pre-analytical, analytical and post-analytical variables that may affect the final test result obtained using DBS specimens used for monitoring of patients with an IMD.

scholarly journals ANALYSIS OF DOXORUBICIN AND DOXORUBICINOL IN DRIED BLOOD SPOT OF BREAST CANCER PATIENTS FOR MONITORING THE CARDIOTOXICITY OF DOXORUBICIN

2020 ◽  
pp. 21-25
Author(s):  
YAHDIANA HARAHAP ◽  
SABRINA NUR AMALIA ◽  
ALDHI ANARTA ◽  
RAMADHAN
Keyword(s):  
Breast Cancer ◽  
Cancer Patients ◽  
Gradient Elution ◽  
Ultra Performance Liquid Chromatography ◽  
Incidence Rates ◽  
Dried Blood Spot ◽  
Precipitation Method ◽  
Breast Cancer Patients ◽  
Acquity Uplc ◽  
Blood Spot

Objective: This study aimed to analyze doxorubicin and doxorubicinol levels in Dried Blood Spot (DBS) from 25 breast cancer patients who received doxorubicin in their therapeutic regiment. Methods: DBS samples were extracted by protein precipitation method and analyzed using Ultra Performance Liquid Chromatography tandem Mass Spectrometry (LC-MS/MS), with the Acquity UPLC BEH C18 Waters chromatography column (2.1 x 100 mm x 1.7 μm). The mobile phase consisted of 0.1% acetic acid (eluent A) and acetonitrile (eluent B) with gradient elution; the flow rate was 0.15 ml/min and runtime, 7 min. This method was linear within the concentration range of 10–200 ng/ml for doxorubicin and 4–100 ng/ml for doxorubicinol. Result: The analysis results showed that doxorubicin levels were in the range of 11.01 ng/ml to 93.75 ng/ml and doxorubicinol was 5.80 ng/ml to 58.57 ng/ml. Conclusion: Cumulative doses of all patients were in the range of 49.11 mg/m2to 303.70 mg/m2, which have cardiomyopathy incidence rates<4%.

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.

Determination of haemoglobin derivatives in aged dried blood spot to estimate haematocrit

2019 ◽  
Vol 57 (7) ◽  
pp. 1026-1034 ◽  
Author(s):  
Rosita Zakaria ◽  
Katrina J. Allen ◽  
Jennifer J. Koplin ◽  
Nick Crinis ◽  
Lidia De Rosa ◽  
...  
Keyword(s):  
Whole Blood ◽  
Method Development ◽  
Evaluation Studies ◽  
Visible Spectrum ◽  
Epidemiological Studies ◽  
Dried Blood Spot ◽  
Therapeutic Drug ◽  
High Positive Correlation ◽  
Low Concentrations ◽  
Blood Spot

Abstract Introduction Dried blood spot (DBS) sample applications now encompass analytes related to clinical diagnosis, epidemiological studies, therapeutic drug monitoring, pharmacokinetic and toxicokinetic studies. Haematocrit (Hct) and haemoglobin (Hb) at very high or low concentrations may influence the accuracy of measurement quantification of the DBS sample. In this study, we aimed to predict the Hct of the punched DBS through primary spectrophotometric estimation of its haemoglobin-derivative (Hb-drv) content. Methods Formic acid solution was used to elute Hb-drv content of 3.2 mm spotted blood from its dry matrix. Direct spectrometry measurement was utilised to scan the extracted Hb-drv in the visible spectrum range of 520–600 nm. The linear relationship between an individual’s Hct percentage and Hb-drv concentration was applied to estimate the Hct level of the blood spot. De-identified whole blood samples were used for the method development and evaluation studies. Results The Hb-drv estimation is valid in samples >2 months old. Method validation experiments DBS demonstrate linearity between 82.5 and 207.5 g/L, average coefficient of variation of 3.6% (intra-assay) and 7.7% (inter-assay), analytical recovery of 84%, and a high positive correlation (r=0.88) between Hb-drv and the original whole blood Hct. The Bland-Altman difference plot demonstrates a mean difference of 2.4% between the calculated DBS Hct and the directly measured Hct from fresh whole bloods. Conclusions We have successfully developed a simple Hb-drv method to estimate Hct in aged DBS samples. This method can be incorporated into DBS analytical work-flow for the in-situ estimation of Hct and subsequent correction of the analyte of interest as required.

scholarly journals Validation of high-throughput, semiquantitative solid phase SARS coronavirus-2 serology assays in serum and dried blood spot matrices

Bioanalysis ◽  
2021 ◽  
Author(s):  
Leo Maritz ◽  
Nicholas J Woudberg ◽  
Amber C Bennett ◽  
Andreia Soares ◽  
Florian Lapierre ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Solid Phase ◽  
Dried Blood Spot ◽  
Sars Coronavirus ◽  
Diagnostic Specificity ◽  
Global Pandemic ◽  
Specificity And Sensitivity ◽  
Infection Monitoring ◽  
Blood Spot

Aim: Serological assays for the detection of anti-SARS coronavirus-2 (SARS-CoV-2) antibodies are essential to the response to the global pandemic. A ligand binding-based serological assay was validated for the semiquantitative detection of IgG, IgM, IgA and neutralizing antibodies (nAb) against SARS-CoV-2 in serum. Results: The assay demonstrated high levels of diagnostic specificity and sensitivity (85–99% for all analytes). Serum IgG, IgM, IgA and nAb correlated positively (R2 = 0.937, R2 = 0.839, R2 = 0.939 and R2 = 0.501, p < 0.001, respectively) with those measured in dried blood spot samples collected using the hemaPEN® microsampling device (Trajan Scientific and Medical, Victoria, Australia). In vitro SARS-CoV-2 pseudotype neutralization correlated positively with the solid phase nAb signals in convalescent donors (R2 = 0.458, p < 0.05). Conclusion: The assay is applicable in efficacy studies, infection monitoring and postmarketing surveillance following vaccine rollout.

scholarly journals SIMULTANEOUS ANALYTICAL METHOD DEVELOPMENT OF 6-MERCAPTOPURINE AND 6-METHYLMERCAPTOPURINE IN DRIED BLOOD SPOT USING ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY

2017 ◽  
Vol 9 ◽  
pp. 168
Author(s):  
Marlina Ika ◽  
Rizka Andalusia ◽  
Supandi Supandi ◽  
Yahdiana Harahap
Keyword(s):  
Liquid Chromatography ◽  
Method Development ◽  
Matrix Effects ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Ultra Performance Liquid Chromatography ◽  
Dried Blood Spot ◽  
Mass Detection ◽  
Relative Standard ◽  
Blood Spot

Objective: 6-mercaptopurine (6-MP) is a chemotherapeutic agent in the antimetabolite class. It has to go through the metabolic pathway to form6-methyl MP (6-MMP). This study aimed to obtain an optimum and validated method for the analysis of 6-MP and 6-MMP in dried blood spot (DBS)samples simultaneously and to evaluate the potential for future drug concentration monitoring in DBS samples.Methods: The quality control and calibration curves were made by spotting 40 μL blood on DBS paper and dried for 3 hrs. DBS papers were cut with adiameter of 8 mm and extracted with acetonitrile-methanol (1:3) containing internal standard 5-fluorouracil (5-FU). Separation was performed withwaters acquity ultra performance liquid chromatography BEH C18 column of 1.7 μm (2.1×100 mm) with a mobile phase consisting of 0.1% formicacid in water 0.1% formic acid in acetonitrile with gradient elution and a flow rate of 0.2 mL/minute. Mass detection was performed using WatersXevo TQD with positive electrospray ionization (ESI) for 6-MP and 6-MMP and negative ESI for 5-FU in the multiple reaction monitoring mode.Results: The detection rates of 6-MP, 6-MMP, and 5-FU were 153.09>119.09, 167.17>126.03, and 129.09>42.05, respectively. This method was linearwith the range at 26-1000 ng/mL for 6-MP and 13-500 ng/mL for 6-MMP with consecutive r≥0.998 and ≥0.999, respectively. The % relative errorvalue and % relative standard deviation for accuracy and precision of intraday and interday were not more than 15% and not more than 20% at thelower limit of quantification concentration, respectively.Conclusions: This method fulfilled the requirements of selectivity, linearity, carry over, and matrix effects referring to the European Medicines Agencyguidelines.

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