Simultaneous determination of tramadol and paracetamol in human plasma using LC-MS/MS and application in bioequivalence study of fixed-dose combination
The study aimed to develop a sensitive and high-throughput liquid chromatography coupled with tandem mass spectrometry method to quantify concentrations of tramadol and paracetamol simultaneously in human plasma. Sample preparation involved single-step protein precipitation using methanol and two de...
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| Vydáno v: | Annals of medicine (Helsinki) Ročník 55; číslo 2; s. 2270502 |
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| Hlavní autoři: | , , , , , , , , , , |
| Médium: | Journal Article |
| Jazyk: | angličtina |
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Taylor & Francis
12.12.2023
Taylor & Francis Group |
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| ISSN: | 0785-3890, 1365-2060, 1365-2060 |
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| Abstract | The study aimed to develop a sensitive and high-throughput liquid chromatography coupled with tandem mass spectrometry method to quantify concentrations of tramadol and paracetamol simultaneously in human plasma. Sample preparation involved single-step protein precipitation using methanol and two deuterated internal standards, tramadol D6 and paracetamol D4. Agilent Poroshell 120 EC-C18 (100 × 2.1 mm, 2.1 µm) analytical column was employed to achieve chromatographic separation. Detection was in positive ion multiple reaction monitoring mode. A tailing factor (Tf) of <1.2, separation factor (K prime) of >1.5 from the column dead time and signal-to-noise (S/N) ratio >10, were obtained for analytes and internal standards. The standard curve was linear over the concentration range of 2.5-500.00 ng/mL for tramadol and 0.025-20.00 μg/mL for paracetamol. A small injection volume of 1 µL, low flow rate of 440 µL/min and short analysis time of 3.5 min reduced the solvent consumption, analysis cost and system contamination. The results of method validation parameters fulfilled the acceptance criteria of bioanalytical guidelines. The method was successfully applied to a bioequivalence study of fixed-dose combination products of tramadol and paracetamol in Malaysian healthy subjects.The study aimed to develop a sensitive and high-throughput liquid chromatography coupled with tandem mass spectrometry method to quantify concentrations of tramadol and paracetamol simultaneously in human plasma. Sample preparation involved single-step protein precipitation using methanol and two deuterated internal standards, tramadol D6 and paracetamol D4. Agilent Poroshell 120 EC-C18 (100 × 2.1 mm, 2.1 µm) analytical column was employed to achieve chromatographic separation. Detection was in positive ion multiple reaction monitoring mode. A tailing factor (Tf) of <1.2, separation factor (K prime) of >1.5 from the column dead time and signal-to-noise (S/N) ratio >10, were obtained for analytes and internal standards. The standard curve was linear over the concentration range of 2.5-500.00 ng/mL for tramadol and 0.025-20.00 μg/mL for paracetamol. A small injection volume of 1 µL, low flow rate of 440 µL/min and short analysis time of 3.5 min reduced the solvent consumption, analysis cost and system contamination. The results of method validation parameters fulfilled the acceptance criteria of bioanalytical guidelines. The method was successfully applied to a bioequivalence study of fixed-dose combination products of tramadol and paracetamol in Malaysian healthy subjects. |
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| AbstractList | The study aimed to develop a sensitive and high-throughput liquid chromatography coupled with tandem mass spectrometry method to quantify concentrations of tramadol and paracetamol simultaneously in human plasma. Sample preparation involved single-step protein precipitation using methanol and two deuterated internal standards, tramadol D6 and paracetamol D4. Agilent Poroshell 120 EC-C18 (100 × 2.1 mm, 2.1 µm) analytical column was employed to achieve chromatographic separation. Detection was in positive ion multiple reaction monitoring mode. A tailing factor (Tf) of <1.2, separation factor (K prime) of >1.5 from the column dead time and signal-to-noise (S/N) ratio >10, were obtained for analytes and internal standards. The standard curve was linear over the concentration range of 2.5–500.00 ng/mL for tramadol and 0.025–20.00 μg/mL for paracetamol. A small injection volume of 1 µL, low flow rate of 440 µL/min and short analysis time of 3.5 min reduced the solvent consumption, analysis cost and system contamination. The results of method validation parameters fulfilled the acceptance criteria of bioanalytical guidelines. The method was successfully applied to a bioequivalence study of fixed-dose combination products of tramadol and paracetamol in Malaysian healthy subjects. AbstractThe study aimed to develop a sensitive and high-throughput liquid chromatography coupled with tandem mass spectrometry method to quantify concentrations of tramadol and paracetamol simultaneously in human plasma. Sample preparation involved single-step protein precipitation using methanol and two deuterated internal standards, tramadol D6 and paracetamol D4. Agilent Poroshell 120 EC-C18 (100 × 2.1 mm, 2.1 µm) analytical column was employed to achieve chromatographic separation. Detection was in positive ion multiple reaction monitoring mode. A tailing factor (Tf) of <1.2, separation factor (K prime) of >1.5 from the column dead time and signal-to-noise (S/N) ratio >10, were obtained for analytes and internal standards. The standard curve was linear over the concentration range of 2.5–500.00 ng/mL for tramadol and 0.025–20.00 μg/mL for paracetamol. A small injection volume of 1 µL, low flow rate of 440 µL/min and short analysis time of 3.5 min reduced the solvent consumption, analysis cost and system contamination. The results of method validation parameters fulfilled the acceptance criteria of bioanalytical guidelines. The method was successfully applied to a bioequivalence study of fixed-dose combination products of tramadol and paracetamol in Malaysian healthy subjects. The study aimed to develop a sensitive and high-throughput liquid chromatography coupled with tandem mass spectrometry method to quantify concentrations of tramadol and paracetamol simultaneously in human plasma. Sample preparation involved single-step protein precipitation using methanol and two deuterated internal standards, tramadol D6 and paracetamol D4. Agilent Poroshell 120 EC-C18 (100 × 2.1 mm, 2.1 µm) analytical column was employed to achieve chromatographic separation. Detection was in positive ion multiple reaction monitoring mode. A tailing factor (Tf) of <1.2, separation factor (K prime) of >1.5 from the column dead time and signal-to-noise (S/N) ratio >10, were obtained for analytes and internal standards. The standard curve was linear over the concentration range of 2.5-500.00 ng/mL for tramadol and 0.025-20.00 μg/mL for paracetamol. A small injection volume of 1 µL, low flow rate of 440 µL/min and short analysis time of 3.5 min reduced the solvent consumption, analysis cost and system contamination. The results of method validation parameters fulfilled the acceptance criteria of bioanalytical guidelines. The method was successfully applied to a bioequivalence study of fixed-dose combination products of tramadol and paracetamol in Malaysian healthy subjects.The study aimed to develop a sensitive and high-throughput liquid chromatography coupled with tandem mass spectrometry method to quantify concentrations of tramadol and paracetamol simultaneously in human plasma. Sample preparation involved single-step protein precipitation using methanol and two deuterated internal standards, tramadol D6 and paracetamol D4. Agilent Poroshell 120 EC-C18 (100 × 2.1 mm, 2.1 µm) analytical column was employed to achieve chromatographic separation. Detection was in positive ion multiple reaction monitoring mode. A tailing factor (Tf) of <1.2, separation factor (K prime) of >1.5 from the column dead time and signal-to-noise (S/N) ratio >10, were obtained for analytes and internal standards. The standard curve was linear over the concentration range of 2.5-500.00 ng/mL for tramadol and 0.025-20.00 μg/mL for paracetamol. A small injection volume of 1 µL, low flow rate of 440 µL/min and short analysis time of 3.5 min reduced the solvent consumption, analysis cost and system contamination. The results of method validation parameters fulfilled the acceptance criteria of bioanalytical guidelines. The method was successfully applied to a bioequivalence study of fixed-dose combination products of tramadol and paracetamol in Malaysian healthy subjects. |
| Author | Pang, Lai Hui Lee, Yi Lin Loh, Gabriel Onn Kit Tan, Yvonne Tze Fung Shahridzo, Siti Halimah Goh, Chen Zhu Peh, Kok Khiang Hermansyah, Andi Wong, Emily Yii Ling Long, Chiau Ming Damenthi, Nair |
| Author_xml | – sequence: 1 givenname: Gabriel Onn Kit surname: Loh fullname: Loh, Gabriel Onn Kit organization: Bioxis Sdn. Bhd., Taman Perindustrian Bukit Minyak, Simpang Ampat, Penang, Malaysia – sequence: 2 givenname: Emily Yii Ling surname: Wong fullname: Wong, Emily Yii Ling organization: Bioxis Sdn. Bhd., Taman Perindustrian Bukit Minyak, Simpang Ampat, Penang, Malaysia – sequence: 3 givenname: Chen Zhu surname: Goh fullname: Goh, Chen Zhu organization: Bioxis Sdn. Bhd., Taman Perindustrian Bukit Minyak, Simpang Ampat, Penang, Malaysia – sequence: 4 givenname: Yvonne Tze Fung surname: Tan fullname: Tan, Yvonne Tze Fung organization: Bioxis Sdn. Bhd., Taman Perindustrian Bukit Minyak, Simpang Ampat, Penang, Malaysia – sequence: 5 givenname: Yi Lin surname: Lee fullname: Lee, Yi Lin organization: Centre for Clinical Trial, Institute for Clinical Research, Ampang Hospital, Ministry of Health, Jalan Mewah Utara, Ampang, Selangor, Malaysia – sequence: 6 givenname: Lai Hui surname: Pang fullname: Pang, Lai Hui organization: Centre for Clinical Trial, Institute for Clinical Research, Ampang Hospital, Ministry of Health, Jalan Mewah Utara, Ampang, Selangor, Malaysia – sequence: 7 givenname: Siti Halimah surname: Shahridzo fullname: Shahridzo, Siti Halimah organization: Centre for Clinical Trial, Institute for Clinical Research, Ampang Hospital, Ministry of Health, Jalan Mewah Utara, Ampang, Selangor, Malaysia – sequence: 8 givenname: Nair surname: Damenthi fullname: Damenthi, Nair organization: Centre for Clinical Trial, Institute for Clinical Research, Ampang Hospital, Ministry of Health, Jalan Mewah Utara, Ampang, Selangor, Malaysia – sequence: 9 givenname: Andi surname: Hermansyah fullname: Hermansyah, Andi organization: Department of Pharmacy Practice, Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia – sequence: 10 givenname: Chiau Ming surname: Long fullname: Long, Chiau Ming organization: Department of Pharmacy Practice, Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia, Pengiran Anak Puteri Rashidah Sa’adatul Bolkiah Institute of Health Sciences, Universiti Brunei Darussalam, Gadong, Brunei Darussalam, School of Medical and Life Sciences, Sunway University, Sunway City, Malaysia – sequence: 11 givenname: Kok Khiang surname: Peh fullname: Peh, Kok Khiang organization: School of Pharmaceutical Sciences, Universiti Sains Malaysia, Minden, Penang, Malaysia |
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| SubjectTerms | bioequivalence study fixed-dose combination LC-ESI-MS/MS method paracetamol Toxicology Tramadol |
| Title | Simultaneous determination of tramadol and paracetamol in human plasma using LC-MS/MS and application in bioequivalence study of fixed-dose combination |
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