Publicação
Development and validation of LC-MS/MS method for determination of fentanyl and midazolam in DBS samples
| Resumo: | Fentanyl and midazolam are two drugs widely used in combination for sedation and analgesic procedures in children. Fentanyl is an analgesic opioid and midazolam is a benzodiazepine of short acting. Caution must be exercised when benzodiazepines and opioids are used together: the risks of hypoxia and apnea are significantly greater than when they are used alone. So, bioanalytical methods with low limit of quantification and high selectivity could be required to control these drugs in hospital environment and particularly in pharmacokinetic studies. In order to improve this necessity, the aim of this work is to develop and try to validate a new method for simultaneous analysis of these two substances in human blood of infants patients under this kind of therapies. The dried blood spots method associated to liquid chromatography tandem mass spectrometry (LC-MS/MS) have been shown a lot of advantages in comparison with the normal techniques. DBS samples were prepared by spotting 20 μL of spiked whole blood onto Protein Saver ™ 903 ™ Whatman ™ cards and drying for at least 2 h. The drug samples were extracted from spots using methanol. Chromatographic separation was performed on a Kinetex 50 mm × 2.1 mm C18 column, using 0,1% formic acid and acetonitrile as mobile phases. The developed method was formed to be accurate, repeatable and selective in the range of linearity from 0,1 to 100 μg/L of fentanyl and from 10 to 4000 μg/L midazolam. The extraction recovery was > 80 % and stability shows good results. However, it was showed a significant matrix effect what mean that a newer confirmation is required. The method was acute, precise and linear and in the future could be successfully used for simultaneous measurements of fentanyl and midazolam concentrations in BDS samples of children patients. |
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| Autores principais: | Azedo, Ana Filipa Freire |
| Assunto: | Fentanyl Midazolam Dried Blood Spots Liquid chromatography tandem mass spectrometry Method validation Mestrado Integrado - 2014 |
| Ano: | 2014 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso restrito |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Fentanyl and midazolam are two drugs widely used in combination for sedation and analgesic procedures in children. Fentanyl is an analgesic opioid and midazolam is a benzodiazepine of short acting. Caution must be exercised when benzodiazepines and opioids are used together: the risks of hypoxia and apnea are significantly greater than when they are used alone. So, bioanalytical methods with low limit of quantification and high selectivity could be required to control these drugs in hospital environment and particularly in pharmacokinetic studies. In order to improve this necessity, the aim of this work is to develop and try to validate a new method for simultaneous analysis of these two substances in human blood of infants patients under this kind of therapies. The dried blood spots method associated to liquid chromatography tandem mass spectrometry (LC-MS/MS) have been shown a lot of advantages in comparison with the normal techniques. DBS samples were prepared by spotting 20 μL of spiked whole blood onto Protein Saver ™ 903 ™ Whatman ™ cards and drying for at least 2 h. The drug samples were extracted from spots using methanol. Chromatographic separation was performed on a Kinetex 50 mm × 2.1 mm C18 column, using 0,1% formic acid and acetonitrile as mobile phases. The developed method was formed to be accurate, repeatable and selective in the range of linearity from 0,1 to 100 μg/L of fentanyl and from 10 to 4000 μg/L midazolam. The extraction recovery was > 80 % and stability shows good results. However, it was showed a significant matrix effect what mean that a newer confirmation is required. The method was acute, precise and linear and in the future could be successfully used for simultaneous measurements of fentanyl and midazolam concentrations in BDS samples of children patients. |
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