Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance
| dc.contributor.advisor | Pillay, Joanne | |
| dc.contributor.advisor | van der Watt, George | |
| dc.contributor.author | van Der Westhuizen, Diederick J | |
| dc.date.accessioned | 2023-02-23T12:16:49Z | |
| dc.date.available | 2023-02-23T12:16:49Z | |
| dc.date.issued | 2022 | |
| dc.date.updated | 2023-02-21T07:26:32Z | |
| dc.description.abstract | The glomerular filtration rate (GFR) is considered the best indicator of kidney function. Iohexol, a contrast agent, is currently considered to be a reference marker since it meets all the requirements of an ideal GFR marker. The aim of this study was to develop and validate a method for iohexol measurement using high performance liquid chromatography with ultraviolet detection (HPLCUV). The method developed includes a mobile phase with water and acetonitrile with a gradient of 5%-30% organic, a C18 analytical column (100 × 4.6 mm, 2.7 µm particle size) at a temperature of 40 °C and a flow rate of 0.5 ml/min. Serum samples were deproteinized by addition of perchloric acid (5%). The assay met the sensitivity cut-off with a mean signal-to-noise ratio of 17.2 at a level of 10 µg/mL iohexol. Mean recovery was 103.7% (CV=4.4%). The dilution experiment allowed for 5-times dilution up to iohexol levels of 500 µg/mL with an accuracy of 103.1% (CV=1.3%) For selectivity, no interfering endogenous compounds at the retention time of iohexol were observed. The matrix effect experiment showed a clinically acceptable variation at all concentration levels with a CV of the slopes of 2.7%. Stability of the stock solution was proven for at least 9 months at -80°C. Sample post-extraction stability was adequate at 84 hours. Interference testing yielded between -7% and -12% difference at 2% haemolysis and -9% to -13% difference at 10% lipaemia. With regards to these findings, this method is simple, specific, linear, precise and robust, which allows its application for the measurement of GFR in serum. | |
| dc.identifier.apacitation | van Der Westhuizen, D. J. (2022). <i>Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance</i>. (). ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences. Retrieved from http://hdl.handle.net/11427/37046 | en_ZA |
| dc.identifier.chicagocitation | van Der Westhuizen, Diederick J. <i>"Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance."</i> ., ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences, 2022. http://hdl.handle.net/11427/37046 | en_ZA |
| dc.identifier.citation | van Der Westhuizen, D.J. 2022. Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance. . ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences. http://hdl.handle.net/11427/37046 | en_ZA |
| dc.identifier.ris | TY - Master Thesis AU - van Der Westhuizen, Diederick J AB - The glomerular filtration rate (GFR) is considered the best indicator of kidney function. Iohexol, a contrast agent, is currently considered to be a reference marker since it meets all the requirements of an ideal GFR marker. The aim of this study was to develop and validate a method for iohexol measurement using high performance liquid chromatography with ultraviolet detection (HPLCUV). The method developed includes a mobile phase with water and acetonitrile with a gradient of 5%-30% organic, a C18 analytical column (100 × 4.6 mm, 2.7 µm particle size) at a temperature of 40 °C and a flow rate of 0.5 ml/min. Serum samples were deproteinized by addition of perchloric acid (5%). The assay met the sensitivity cut-off with a mean signal-to-noise ratio of 17.2 at a level of 10 µg/mL iohexol. Mean recovery was 103.7% (CV=4.4%). The dilution experiment allowed for 5-times dilution up to iohexol levels of 500 µg/mL with an accuracy of 103.1% (CV=1.3%) For selectivity, no interfering endogenous compounds at the retention time of iohexol were observed. The matrix effect experiment showed a clinically acceptable variation at all concentration levels with a CV of the slopes of 2.7%. Stability of the stock solution was proven for at least 9 months at -80°C. Sample post-extraction stability was adequate at 84 hours. Interference testing yielded between -7% and -12% difference at 2% haemolysis and -9% to -13% difference at 10% lipaemia. With regards to these findings, this method is simple, specific, linear, precise and robust, which allows its application for the measurement of GFR in serum. DA - 2022_ DB - OpenUCT DP - University of Cape Town KW - Medicine LK - https://open.uct.ac.za PY - 2022 T1 - Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance TI - Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance UR - http://hdl.handle.net/11427/37046 ER - | en_ZA |
| dc.identifier.uri | http://hdl.handle.net/11427/37046 | |
| dc.identifier.vancouvercitation | van Der Westhuizen DJ. Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance. []. ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences, 2022 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/37046 | en_ZA |
| dc.language.rfc3066 | eng | |
| dc.publisher.department | Department of Clinical Laboratory Sciences | |
| dc.publisher.faculty | Faculty of Health Sciences | |
| dc.subject | Medicine | |
| dc.title | Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance | |
| dc.type | Master Thesis | |
| dc.type.qualificationlevel | Masters | |
| dc.type.qualificationlevel | MMed |