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1.
Bioanalysis ; 11(21): 1941-1954, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31829054

RESUMEN

Aim: Quantitative LC-MS analysis of oligonucleotides (OGNs) in biological matrices is needed to support candidate selection of new therapeutic OGNs. Methodology & results: A set of 20 single stranded antisense oligonucleotides (ASO) and five siRNAs were extracted from plasma and tissue homogenates. Anion Exchange (AEX) SPE was selected as generic extraction approach, resulting in recoveries from plasma >70%. Extraction from tissue homogenates showed often more variation and lower recoveries. A proof of concept of a novel tailored hybridization extraction is demonstrated for two 16-mer reference OGNs. Conclusion: Two methods for extraction of OGNs were investigated and applied for quantitative analysis. The AEX-SPE is considered a more generic approach preferred when multiple compounds are evaluated. Hybridization extraction has great potential but critical reagents per analyte are needed.


Asunto(s)
Cromatografía Liquida/métodos , Espectrometría de Masas/métodos , Oligonucleótidos/análisis , Oligonucleótidos/aislamiento & purificación , Extracción en Fase Sólida/métodos , Secuencia de Bases , Humanos , Límite de Detección , Hibridación de Ácido Nucleico , Oligonucleótidos/sangre , Oligonucleótidos/genética
2.
Bioanalysis ; 9(23): 1859-1872, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29205051

RESUMEN

AIM: Imetelstat, a 13-mer oligonucleotide with a lipid tail is being evaluated for treating hematologic myeloid malignancies. This report describes the development of extraction and quantification methods for imetelstat. Methodology & results: Imetelstat was extracted using SPE (rat plasma) or by hybridization using a biotinylated capture probe (human plasma) and was quantified by LC-MS/MS. Calibration curves were established (0.1-50 µg/ml). Stability of imetelstat in plasma was demonstrated. Concentrations of imetelstat extracted using either of the methods and quantified with LC-MS/MS were comparable with a validated ELISA. CONCLUSION: Two extraction methods (solid phase and hybridization) were developed for quantifying imetelstat in plasma using LC-MS/MS. The hybridization extraction in combination with LC-MS/MS is a novel extraction approach.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Oligonucleótidos/sangre , Espectrometría de Masas en Tándem/métodos , Animales , Calibración , Cromatografía Líquida de Alta Presión/normas , Sondas de ADN/metabolismo , Ensayo de Inmunoadsorción Enzimática , Semivida , Humanos , Límite de Detección , Masculino , Hibridación de Ácido Nucleico , Oligonucleótidos/aislamiento & purificación , Oligonucleótidos/metabolismo , Ratas , Ratas Sprague-Dawley , Extracción en Fase Sólida , Espectrometría de Masas en Tándem/normas
3.
Bioanalysis ; 8(19): 2007-21, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27611836

RESUMEN

AIM: A procedure was developed for the assessment of photochemical stability of drugs in blood and plasma under standardized conditions. The procedure avoids a variable outcome of photochemical stability experiments and tests relevant worst case conditions so that unnecessary light protection is avoided. Results/methodology: Blood and plasma were spiked with a mixture of drugs and incubated in a Suntest CPS(+), in the laboratory on the bench and near the window on a sunny summer day. The results were compared. DISCUSSION/CONCLUSION: No protection from light, limited protection from light and full protection from light are advised for drugs that are stable in plasma in the Suntest CPS(+) at 250 W/m(2) for at least 30 min, for 5-30 min and for <5 min, respectively.


Asunto(s)
Estabilidad de Medicamentos , Luz , Preparaciones Farmacéuticas/sangre , Preparaciones Farmacéuticas/efectos de la radiación , Cromatografía Líquida de Alta Presión , Humanos , Fotólisis/efectos de la radiación , Espectrometría de Masas en Tándem , Rayos Ultravioleta
4.
Bioanalysis ; 7(20): 2713-24, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26507928

RESUMEN

Ronald de Vries graduated in Organic and Analytical Chemistry at the Free University of Amsterdam, The Netherlands. After working in a Contract Laboratory (CRO) for 7 years, he joined Janssen R&D in 1998. At Janssen R&D, Belgium, Ronald worked in the bioanalytical department that supports both clinical and nonclinical bioanalysis. In this department he had several roles, such as providing the bioanalytical support for various drug development programs and leading the method establishment group. He has done numerous global assay transfers to/from Janssen from/to other laboratories and plays an important role in the introduction and application of new technologies and applied innovation in the department. In 2014 he started in the drug metabolism and pharmacokinetics department of Janssen R&D, where his main tasks are in vivo and in vitro metabolite identification using high resolution MS and Radiodetection.


Asunto(s)
Análisis Químico de la Sangre/métodos , Pirazoles/sangre , Pirimidinas/sangre , Adenina/análogos & derivados , Agammaglobulinemia Tirosina Quinasa , Ácidos y Sales Biliares/química , Análisis Químico de la Sangre/normas , Calibración , Cromatografía Líquida de Alta Presión/normas , Humanos , Hepatopatías/metabolismo , Hepatopatías/patología , Piperidinas , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Proteínas Tirosina Quinasas/metabolismo , Pirazoles/metabolismo , Pirimidinas/metabolismo , Control de Calidad , Espectrometría de Masas en Tándem/normas , Ácido Taurocólico/química
5.
Artículo en Inglés | MEDLINE | ID: mdl-25600054

RESUMEN

Tapentadol is a novel, centrally acting analgesic with 2 mechanisms of action, MOR agonism and noradrenaline (NA) reuptake inhibition in a single molecule. It is the first member of a new therapeutic class, MOR-NRI. A high throughput liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay was developed and validated for the quantitative analysis of tapentadol and its O-glucuronide metabolite in human serum. Simultaneous quantification was deemed to be challenging because of the large difference in concentrations between tapentadol and its O-glucuronide metabolite in clinical samples. Therefore, a method was established using a common processed sample, but with different injection volumes and chromatographic conditions for each analyte. Tapentadol and tapentadol-O-glucuronide were determined by protein precipitation of 0.100ml of the samples with acetonitrile. The internal standards used are D6-tapentadol and D6-tapentadol-O-glucuronide. The validated concentration range was 0.200-200 ng/ml (tapentadol) and 10.0-10,000 ng/ml (tapentadol-O-glucuronide). Chromatographic separation was achieved by gradient elution on a Waters Acquity UPLC BEH C18 (1.7 µm, 2.1 × 50 mm) column, with mobile phase consisting of 0.01 M ammonium formate (adjusted to pH 4 using formic acid) (A) and methanol (B). A separate injection was done for measurement of each analyte, with a different gradient and run time. The analytes were detected by using an electrospray ion source on a triple quadrupole mass spectrometer operating in positive ionization mode. The run time was 1.6 min for tapentadol and 1.5 min for tapentadol-O-glucuronide. The high sensitivity and acceptable performance of the assay allowed its application to the analysis of serum samples in clinical trials. The validated method was used for analysis of tapentadol in over 17,000 samples.


Asunto(s)
Analgésicos/sangre , Cromatografía Líquida de Alta Presión/métodos , Glucurónidos/sangre , Fenoles/sangre , Espectrometría de Masas en Tándem/métodos , Analgésicos/administración & dosificación , Analgésicos/farmacocinética , Analgésicos/uso terapéutico , Humanos , Modelos Lineales , Neoplasias/complicaciones , Dolor/tratamiento farmacológico , Dolor/etiología , Fenoles/administración & dosificación , Fenoles/farmacocinética , Fenoles/uso terapéutico , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Tapentadol
6.
Artículo en Inglés | MEDLINE | ID: mdl-18571483

RESUMEN

Two liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) methods are described, one for the quantitative determination of risperidone and the enantiomers of its active metabolite 9-hydroxyrisperidone (paliperidone) in human plasma and the other for the determination of the enantiomers of 9-hydroxyrisperidone in human urine. The plasma method is based on solid-phase extraction of 200 microl of sample on a mixed-mode sorbent, followed by separation on a cellulose-based LC column with a 13.5-min mobile phase gradient of hexane, isopropanol and ethanol. After post-column addition of 10 mM ammonium acetate in ethanol/water, detection takes place by ion-spray tandem mass spectrometry in the positive ion mode. Method validation results show that the method is sufficiently selective towards the enantiomers of 7-hydroxyrisperidone and capable of quantifying the analytes with good precision and accuracy in the concentration range of 0.2-100 ng/ml. An accelerated (run time of 4.3 min) and equally valid method for the enantiomers of 9-hydroxyrisperidone alone in plasma is obtained by increasing the mobile phase flow-rate from 1.0 to 2.0 ml/min and slightly adapting the gradient conditions. The urine method is based on the same solid-phase extraction and chromatographic approach as the accelerated plasma method. Using 100 microl of sample, (+)- and (-)-9-hydroxyrisperidone can be quantified in the concentration range 1-2000 ng/ml. The accelerated method for plasma and the method for urine can be used only when paliperidone is administered instead of risperidone, as there is insufficient separation of the 9-hydroxy enantiomers from the 7-hydroxy enantiomers, the latter ones being present only after risperidone administration.


Asunto(s)
Cromatografía Liquida/métodos , Isoxazoles/química , Pirimidinas/química , Risperidona/química , Espectrometría de Masas en Tándem/métodos , Humanos , Isoxazoles/sangre , Isoxazoles/farmacocinética , Isoxazoles/orina , Palmitato de Paliperidona , Pirimidinas/sangre , Pirimidinas/farmacocinética , Pirimidinas/orina , Risperidona/sangre , Risperidona/farmacocinética , Risperidona/orina , Sensibilidad y Especificidad
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