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03097nam a22003377a 4500 |
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0299242 |
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|a eng
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|9 396613
|a Al Selmi, Turki Faris
|e Author
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|a Establishment and validation of a clinical biochemistry HPLC-tandem mass spectrometry method for 25-OH-Vitamin D3
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|a ملبورن
|c 201?
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|a 1 - 48
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|a رسائل جامعية
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| 502 |
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|b رسالة ماجستير
|c RMIT University
|f School of Health Science
|g أستراليا
|o 0002
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|a Introduction Clinical interest in vitamin D has resulted in a barrage of tests generated for the assessment of serum 25-hydroxyvitamin D (25(OH)D). This in turn has prompted intense discussion and research into the merits of vitamin D assays. The two over-arching methodologies for vitamin D quantification are chromatography and immunoassay. The aim of this project was to develop a 12 week student practical comparing vitamin D by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Methods A routine LC-MS/MS method for 25(OH)D was developed on the Agilent 1200HPLC/6410QQQ. 25(OH)D3-d6 (hexadeuterated) was used as the internal standard for 25(OH)D3, epi-25(OH)D3 and 25(OH)D2. Multi-level calibrators were purchased from Recipe. Samples were prepared by liquid extraction and eluted by gradient separation on a Varian Pursuit-XRs (20mm x 2.0mm x 3µm) C18 column. Electrospray ionization in the positive ion multiple reaction monitoring mode was used to quantify 25(OH)D3 (401.2/383.3), epi-25(OH)D3 (401.2/383.3), 25(OH)D3-d6 (407.2/389.2) and 25(OH)D2 (413.2/395.5). Results 25(OH)D3 TMS method validation studies demonstrated: within-run imprecision (CV%) of 4.9% and 5.5% at 55nmol/L and 212nmol/L respectively, between-run imprecision of 5.0% and 5.2% at 25nmol/L and 75nmol/L respectively, linearity to at least 212nmol/L, limit of quantitation at 5nmol/L, recovery of 101%, and average bias of 3.2% compared to QAP targets. Conclusions This student project draws together the themes of assay design, development and method validation as students develop skills in both chromatography and immunoassay methods, using vitamin D as an example. This is the first such student project in Australasia.
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|a الكيمياء الحيوية السريرية
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| 653 |
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|a الكيمياء الحيوية
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| 700 |
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|9 45461
|a Roche, Peter
|e Advisor
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|9 396616
|a Greaves, Ronda
|e Advisor
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| 856 |
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|u 9843-001-013-0002-T.pdf
|y صفحة العنوان
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| 856 |
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|u 9843-001-013-0002-A.pdf
|y المستخلص
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| 856 |
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|u 9843-001-013-0002-C.pdf
|y قائمة المحتويات
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| 856 |
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|u 9843-001-013-0002-F.pdf
|y 24 صفحة الأولى
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| 856 |
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|u 9843-001-013-0002-1.pdf
|y 1 الفصل
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| 856 |
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|u 9843-001-013-0002-2.pdf
|y 2 الفصل
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| 856 |
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|u 9843-001-013-0002-3.pdf
|y 3 الفصل
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| 856 |
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|u 9843-001-013-0002-O.pdf
|y الخاتمة
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| 856 |
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|u 9843-001-013-0002-R.pdf
|y المصادر والمراجع
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| 856 |
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|u 9843-001-013-0002-S.pdf
|y الملاحق
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| 930 |
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|d n
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|a +Dissertations
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|c 752564
|d 752564
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