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Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance
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 mea...
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| Format: | Thesis |
| Language: | English |
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Department of Clinical Laboratory Sciences
2023
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| _version_ | 1867613326547091456 |
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| access_status_str | Open Access |
| author | van Der Westhuizen, Diederick J |
| author2 | Pillay, Joanne |
| author_browse | Pillay, Joanne van Der Westhuizen, Diederick J |
| author_facet | Pillay, Joanne van Der Westhuizen, Diederick J |
| author_sort | van Der Westhuizen, Diederick J |
| collection | Thesis |
| description | 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. |
| format | Thesis |
| id | oai:open.uct.ac.za:11427/37046 |
| institution | University of Cape Town (South Africa) |
| language | eng |
| last_indexed | 2026-06-10T12:34:20.437Z |
| license_str | Not specified — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | Department of Clinical Laboratory Sciences |
| publisherStr | Department of Clinical Laboratory Sciences |
| record_format | dspace |
| source_str | UCTD — University of Cape Town Open Access Repository |
| spelling | oai:open.uct.ac.za:11427/37046 Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance van Der Westhuizen, Diederick J Pillay, Joanne van der Watt, George Medicine 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. 2023-02-23T12:16:49Z 2023-02-23T12:16:49Z 2022 2023-02-21T07:26:32Z Master Thesis Masters MMed http://hdl.handle.net/11427/37046 eng application/pdf Department of Clinical Laboratory Sciences Faculty of Health Sciences |
| spellingShingle | Medicine van Der Westhuizen, Diederick J Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance |
| thesis_degree_str | Master's |
| title | Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance |
| title_full | Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance |
| title_fullStr | Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance |
| title_full_unstemmed | Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance |
| title_short | Development of a simple HPLC-UV method for determination of GFR by serum iohexol clearance |
| title_sort | development of a simple hplc uv method for determination of gfr by serum iohexol clearance |
| topic | Medicine |
| url | http://hdl.handle.net/11427/37046 |
| work_keys_str_mv | AT vanderwesthuizendiederickj developmentofasimplehplcuvmethodfordeterminationofgfrbyserumiohexolclearance |