Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children
We have developed and validated a novel, sensitive, selective and reproducible reversed-phase high-performance liquid chromatography method coupled with electrospray ionization mass spectrometry (HPLC–ESI-MS/MS) for the simultaneous quantitation of ceftriaxone (CEF), metronidazole (MET) and hydroxym...
| Príomhchruthaitheoirí: | , , , , , |
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| Formáid: | Journal article |
| Foilsithe / Cruthaithe: |
Wellcome Trust
2017
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| Ábhair: |
| _version_ | 1826268302551285760 |
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| author | Ongas, M Standing, J Ogutu, B Waichungo, J Berkley, J Kipper, K |
| author_facet | Ongas, M Standing, J Ogutu, B Waichungo, J Berkley, J Kipper, K |
| author_sort | Ongas, M |
| collection | OXFORD |
| description | We have developed and validated a novel, sensitive, selective and reproducible reversed-phase high-performance liquid chromatography method coupled with electrospray ionization mass spectrometry (HPLC–ESI-MS/MS) for the simultaneous quantitation of ceftriaxone (CEF), metronidazole (MET) and hydroxymetronidazole (MET-OH) from only 50 μL of human plasma, and unbound CEF from 25 μL plasma ultra-filtrate to evaluate the effect of protein binding. Cefuroxime axetil (CEFU) was used as an internal standard (IS). The analytes were extracted by a protein precipitation procedure with acetonitrile and separated on a reversed-phase Polaris 5 C18-Analytical column using a mobile phase composed of acetonitrile containing 0.1% (v/v) formic acid and 10 mM aqueous ammonium formate pH 2.5, delivered at a flow-rate of 300 μL/min. Multiple reaction monitoring was performed in the positive ion mode using the transitions m/z555.1→m/z396.0 (CEF), m/z172.2→m/z 128.2 (MET), m/z188.0→m/z125.9 (MET-OH) and m/z528.1→m/z 364.0 (CEFU) to quantify the drugs. Calibration curves in spiked plasma and ultra-filtrate were linear (r ≥ 0.9948) from 0.4–300 μg/mL for CEF, 0.05–50 μg/mL for MET and 0.02 – 30 μg/mL for MET-OH. The intra- and inter- assay precisions were less than 9% and the mean extraction recoveries were 94.0% (CEF), 98.2% (MET), 99.6% (MET-OH) and 104.6% (CEF in ultra-filtrate); the recoveries for the IS were 93.8% (in plasma) and 97.6% (in ultra-filtrate). The validated method was successfully applied to a pharmacokinetic study of CEF, MET and MET-OH in hospitalized children with complicated severe acute malnutrition following an oral administration of MET and intravenous administration of CEF over the course of 72 hours. |
| first_indexed | 2024-03-06T21:07:36Z |
| format | Journal article |
| id | oxford-uuid:3d020626-f73c-4113-a914-d0540861a86e |
| institution | University of Oxford |
| last_indexed | 2024-03-06T21:07:36Z |
| publishDate | 2017 |
| publisher | Wellcome Trust |
| record_format | dspace |
| spelling | oxford-uuid:3d020626-f73c-4113-a914-d0540861a86e2022-03-26T14:17:00ZLiquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished childrenJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:3d020626-f73c-4113-a914-d0540861a86e*subject*Symplectic Elements at OxfordWellcome Trust2017Ongas, MStanding, JOgutu, BWaichungo, JBerkley, JKipper, KWe have developed and validated a novel, sensitive, selective and reproducible reversed-phase high-performance liquid chromatography method coupled with electrospray ionization mass spectrometry (HPLC–ESI-MS/MS) for the simultaneous quantitation of ceftriaxone (CEF), metronidazole (MET) and hydroxymetronidazole (MET-OH) from only 50 μL of human plasma, and unbound CEF from 25 μL plasma ultra-filtrate to evaluate the effect of protein binding. Cefuroxime axetil (CEFU) was used as an internal standard (IS). The analytes were extracted by a protein precipitation procedure with acetonitrile and separated on a reversed-phase Polaris 5 C18-Analytical column using a mobile phase composed of acetonitrile containing 0.1% (v/v) formic acid and 10 mM aqueous ammonium formate pH 2.5, delivered at a flow-rate of 300 μL/min. Multiple reaction monitoring was performed in the positive ion mode using the transitions m/z555.1→m/z396.0 (CEF), m/z172.2→m/z 128.2 (MET), m/z188.0→m/z125.9 (MET-OH) and m/z528.1→m/z 364.0 (CEFU) to quantify the drugs. Calibration curves in spiked plasma and ultra-filtrate were linear (r ≥ 0.9948) from 0.4–300 μg/mL for CEF, 0.05–50 μg/mL for MET and 0.02 – 30 μg/mL for MET-OH. The intra- and inter- assay precisions were less than 9% and the mean extraction recoveries were 94.0% (CEF), 98.2% (MET), 99.6% (MET-OH) and 104.6% (CEF in ultra-filtrate); the recoveries for the IS were 93.8% (in plasma) and 97.6% (in ultra-filtrate). The validated method was successfully applied to a pharmacokinetic study of CEF, MET and MET-OH in hospitalized children with complicated severe acute malnutrition following an oral administration of MET and intravenous administration of CEF over the course of 72 hours. |
| spellingShingle | *subject* Ongas, M Standing, J Ogutu, B Waichungo, J Berkley, J Kipper, K Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children |
| title | Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children |
| title_full | Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children |
| title_fullStr | Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children |
| title_full_unstemmed | Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children |
| title_short | Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children |
| title_sort | liquid chromatography tandem mass spectrometry for the simultaneous quantitation of ceftriaxone metronidazole and hydroxymetronidazole in plasma from seriously ill severely malnourished children |
| topic | *subject* |
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