Abstract
5-Fluorouracil (5-FU) is a major chemotherapy drug used for the treatment of tumors. It is catabolized mainly by dihydropyrimidine dehydrogenase, and patients with a complete or partial deficiency of dihydropyrimidine dehydrogenase activity are at risk of developing severe 5-FU-associated toxicity. The aim of this study was to demonstrate that intact peripheral blood mononuclear cells (PBMCs) can be an effective model to evaluate the degradation rate of 5-FU. We developed a sensitive and specific liquid chromatography-tandem mass spectrometry method to measure in vitro the rate of 5-FU degradation by intact PBMC. 5-FU degradation rate was determined by measuring the decrease of a fixed amount of 5-FU (10 μg/mL) added to a solution of PBMC, after 2 hours incubation, expressed as nanogram per milliliter of 5-FU degraded per minute × 10 cells. Freshly prepared intact PBMC can degrade efficiently in vitro-added 5-FU. The assay consists of 3 steps: (1) PBMC isolation from peripheral blood, (2) PBMC incubation with 5-FU in vitro for different times, and (3) determination of 5-FU amount to calculate the degradation rate. 5-FU was analyzed by a Q Trap 2000 triple quadrupole/ion trap mass spectrometer in the multiple-reaction-monitoring modes. The chromatographic separation was accomplished using a C18 column with a run time of 16 minutes. By analyzing samples from 39 patients with no 5-FU toxicity, the mean 5-FU degradation rate was 1.85 ± 0.50 ng•mL•min × 10 cells. The assessment of a test to measure 5-FU degradation rate in PBMC of patients before 5-FU administration could represent a prescreening method for evaluating the possible toxicity of this drug as an aid to set up a personalized medicine approach for each patient.
| Original language | English |
|---|---|
| Pages (from-to) | 482-488 |
| Number of pages | 7 |
| Journal | Therapeutic Drug Monitoring |
| Volume | 31 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Aug 2009 |
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Keywords
- 5-fluorouracil
- Dihydropyrimidine dehydrogenase
- LC-MS/MS
- Peripheral blood mononuclear cells, therapeutic drug monitoring
- Personalized medicine
ASJC Scopus subject areas
- Pharmacology (medical)
- Pharmacology
Cite this
A liquid chromatography-tandem mass spectrometry method for the determination of 5-fluorouracil degradation rate by intact peripheral blood mononuclear cells. / Lostia, Alfonso M.; Lionetto, Luana; Ialongo, Cristiano; Gentile, Giovanna; Viterbo, Antonella; Malaguti, Paola; Paris, Ida; Marchetti, Luca; Marchetti, Paolo; Blasi, Antonio De; Simmaco, Maurizio.
In: Therapeutic Drug Monitoring, Vol. 31, No. 4, 08.2009, p. 482-488.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A liquid chromatography-tandem mass spectrometry method for the determination of 5-fluorouracil degradation rate by intact peripheral blood mononuclear cells
AU - Lostia, Alfonso M.
AU - Lionetto, Luana
AU - Ialongo, Cristiano
AU - Gentile, Giovanna
AU - Viterbo, Antonella
AU - Malaguti, Paola
AU - Paris, Ida
AU - Marchetti, Luca
AU - Marchetti, Paolo
AU - Blasi, Antonio De
AU - Simmaco, Maurizio
PY - 2009/8
Y1 - 2009/8
N2 - 5-Fluorouracil (5-FU) is a major chemotherapy drug used for the treatment of tumors. It is catabolized mainly by dihydropyrimidine dehydrogenase, and patients with a complete or partial deficiency of dihydropyrimidine dehydrogenase activity are at risk of developing severe 5-FU-associated toxicity. The aim of this study was to demonstrate that intact peripheral blood mononuclear cells (PBMCs) can be an effective model to evaluate the degradation rate of 5-FU. We developed a sensitive and specific liquid chromatography-tandem mass spectrometry method to measure in vitro the rate of 5-FU degradation by intact PBMC. 5-FU degradation rate was determined by measuring the decrease of a fixed amount of 5-FU (10 μg/mL) added to a solution of PBMC, after 2 hours incubation, expressed as nanogram per milliliter of 5-FU degraded per minute × 10 cells. Freshly prepared intact PBMC can degrade efficiently in vitro-added 5-FU. The assay consists of 3 steps: (1) PBMC isolation from peripheral blood, (2) PBMC incubation with 5-FU in vitro for different times, and (3) determination of 5-FU amount to calculate the degradation rate. 5-FU was analyzed by a Q Trap 2000 triple quadrupole/ion trap mass spectrometer in the multiple-reaction-monitoring modes. The chromatographic separation was accomplished using a C18 column with a run time of 16 minutes. By analyzing samples from 39 patients with no 5-FU toxicity, the mean 5-FU degradation rate was 1.85 ± 0.50 ng•mL•min × 10 cells. The assessment of a test to measure 5-FU degradation rate in PBMC of patients before 5-FU administration could represent a prescreening method for evaluating the possible toxicity of this drug as an aid to set up a personalized medicine approach for each patient.
AB - 5-Fluorouracil (5-FU) is a major chemotherapy drug used for the treatment of tumors. It is catabolized mainly by dihydropyrimidine dehydrogenase, and patients with a complete or partial deficiency of dihydropyrimidine dehydrogenase activity are at risk of developing severe 5-FU-associated toxicity. The aim of this study was to demonstrate that intact peripheral blood mononuclear cells (PBMCs) can be an effective model to evaluate the degradation rate of 5-FU. We developed a sensitive and specific liquid chromatography-tandem mass spectrometry method to measure in vitro the rate of 5-FU degradation by intact PBMC. 5-FU degradation rate was determined by measuring the decrease of a fixed amount of 5-FU (10 μg/mL) added to a solution of PBMC, after 2 hours incubation, expressed as nanogram per milliliter of 5-FU degraded per minute × 10 cells. Freshly prepared intact PBMC can degrade efficiently in vitro-added 5-FU. The assay consists of 3 steps: (1) PBMC isolation from peripheral blood, (2) PBMC incubation with 5-FU in vitro for different times, and (3) determination of 5-FU amount to calculate the degradation rate. 5-FU was analyzed by a Q Trap 2000 triple quadrupole/ion trap mass spectrometer in the multiple-reaction-monitoring modes. The chromatographic separation was accomplished using a C18 column with a run time of 16 minutes. By analyzing samples from 39 patients with no 5-FU toxicity, the mean 5-FU degradation rate was 1.85 ± 0.50 ng•mL•min × 10 cells. The assessment of a test to measure 5-FU degradation rate in PBMC of patients before 5-FU administration could represent a prescreening method for evaluating the possible toxicity of this drug as an aid to set up a personalized medicine approach for each patient.
KW - 5-fluorouracil
KW - Dihydropyrimidine dehydrogenase
KW - LC-MS/MS
KW - Peripheral blood mononuclear cells, therapeutic drug monitoring
KW - Personalized medicine
UR - http://www.scopus.com/inward/record.url?scp=68449102453&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=68449102453&partnerID=8YFLogxK
U2 - 10.1097/FTD.0b013e3181ae4516
DO - 10.1097/FTD.0b013e3181ae4516
M3 - Article
C2 - 19571774
AN - SCOPUS:68449102453
VL - 31
SP - 482
EP - 488
JO - Therapeutic Drug Monitoring
JF - Therapeutic Drug Monitoring
SN - 0163-4356
IS - 4
ER -