VITAMIN B1 & B6 VITAMIN B1&B6 COMBI-DETERMINATION IN EDTA- OF HEPARINE WHOLE BLOOD. HPLC & UPLC Method
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1 VITAMIN B1 & B6 VITAMIN B1&B6 COMBI-DETERMINATION IN EDTA- OF HEPARINE WHOLE BLOOD Method HPLC Vitamin B6 (PLP) min. Internal Standard min. Vitamin B1 (TDP) min. UPLC Vitamin B6 (PLP) min. Internal Standard min. Vitamin B1 (TDP) min. one mobile phase one wavelength (Ex, Em) isocratic method no incubation at 60 o C fluorescence detection 3 levels Calibrator set 3 levels Controle set Product name Product nr. Contents Package Reagent Set - HPLC tests Reagent Set - UPLC tests Direct Calibrator x 2 ml Calibrator Set x 1 ml Control Low Level x 2 ml Control Normal Level x 2 ml Control High Level x 2 ml INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 1 van 12
2 VITAMINE B1 & B6 COMBI - DETERMINATION IN EDTA of HEPARINE WHOLE BLOOD MONSTERMATERIAAL: Whole blood patient sample with EDTA or Heparine HPLC - ANALYTICAL CONDITIONS: Column : XBridge BEH C18 Column, 5 µm, 3.0 x 100 mm Flow Rate : 1.3 ml/min, Mobile Phase Temperature : 30 o C Detection : Fluorescence (ex. 370 nm, em. 455 nm) Injection volume : 35 µl 40 µl Retention times : Vitamin B6 (PLP) ca. 1.8 min. : Internal Standard ca. 5.7 min. : Vitamin B1 (TDP) ca. 7.9 min. UPLC - ANALYTICAL CONDITION: Column : Acquity UPLC BEH C18 Column, 1.7 µm, 2.1 x 30 mm Flow Rate : 0.7 ml/min, Mobile Phase Temperature : 30 o C Detection : Fluorescence (ex. 370 nm, em. 455 nm) Injection volume : 10 µl Retention time : Vitamin B6 (PLP) ca. 0.6 min. : Internal Standard ca. 1.7 min. : Vitamin B1 (TDP) ca. 2.9 min. EXPECTED VALUES: TDP B1 : nmol/l PLP B6 : nmol/l PERFORMANCE: Linearity TDP B1 : tot nmol/l Linearity PLP B6 : tot nmol/l Within run precision TDP B1: N Sample material S.D. CV (%) 25 Whole blood patient samples with EDTA Mean = nmol/l Within run precision PLP B6: N Sample material S.D. CV (%) 25 Whole blood patient samples with EDTA Mean = 69.7 nmol/l INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 2 van 12
3 Calibration line Vitamine B6 & B1 Combi - Determination -1- Calibrator Set (3111) - Vitamin B1 Calibrator 1 = 50 nmol / liter - Vitamin B1 Calibrator 2 = 150 nmol / liter - Vitamin B1 Calibrator 3 = 250 nmol / liter - Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 3 van 12
4 Calibration line Vitamine B6 & B1 Combi - Determination -2- Calibrator Set (3111) - Vitamin B6 Calibrator 1 = 30 nmol / liter - Vitamin B6 Calibrator 2 = 80 nmol / liter - Vitamin B6 Calibrator 3 = 200 nmol / liter - Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 4 van 12
5 Chromatogram Vitamin B6 & B1-3- Direct Calibrator (3006) - Vitamin B6 = 189 nmol / liter - Vitamin B6 = 1.8 min. - Internal Standard - Internal Standard = 5.7 min. - Vitamin B1 = 109 nmol / liter - Vitamin B1 = 7.9 min. HPLC-Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 5 van 12
6 Chromatogram Vitamin B6 & B1 Combi - Determination -4- Calibrator 1 (3108) - Vitamin B6 = 30 nmol / liter - Vitamin B6 = 1.8 min. - Internal Standard - Internal Standard = 5.7 min. - Vitamin B1 = 50 nmol / liter - Vitamin B1 = 7.9 min. HPLC-Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 6 van 12
7 Chromatogram Vitamin B6 & B1 Combi - Determination -5- Calibrator 2 (3109) - Vitamin B6 = 80 nmol / liter - Vitamin B6 = 1.8 min. - Internal Standard - Internal Standard = 5.7 min. - Vitamin B1 = 150 nmol / liter - Vitamin B1 = 7.9 min. HPLC-Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 7 van 12
8 Chromatogram Vitamin B6 & B1 Combi - Determination -6- Calibrator 3 (3110) - Vitamin B6 = 200 nmol / liter - Vitamin B6 = 1.8 min. - Internal Standard - Internal Standard = 5.6 min. - Vitamin B1 = 250 nmol / liter - Vitamin B1 = 7.9 min. HPLC-Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 8 van 12
9 Chromatogram Vitamin B6 & B1-7- Direct Calibrator (3006) - Vitamin B6 = 189 nmol / liter - Vitamin B6 = 0.6 min. - Internal Standard - Internal Standard = 1.7 min. - Vitamin B1 = 109 nmol / liter - Vitamin B1 = 2.9 min. UPLC-Method Vraag meer informatie en/of testverpakking aan. INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 9 van 12
10 Chromatogram Vitamin B6 & B1 Combi - Determination -8- Calibrator 1 (3108) - Vitamin B6 = 30 nmol / liter - Vitamin B6 = 0.6 min. - Internal Standard - Internal Standard = 1.7 min. - Vitamin B1 = 50 nmol / liter - Vitamin B1 = 2.9 min. UPLC-Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 10 van 12
11 Chromatogram Vitamin B6 & B1 Combi - Determination -9- Calibrator 2 (3109) - Vitamin B6 = 80 nmol / liter - Vitamin B6 = 0.6 min. - Internal Standard - Internal Standard = 1.7 min. - Vitamin B1 = 150 nmol / liter - Vitamin B1 = 2.9 min. UPLC-Method INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 11 van 12
12 Chromatogram Vitamin B6 & B1-10- Calibrator 3 (3110) - Vitamin B6 = 200 nmol / liter - Vitamin B6 = 0.6 min. - Internal Standard - Internal Standard = 1.7 min. - Vitamin B1 = 250 nmol / liter - Vitamin B1 = 2.9 min. UPLC-Method Reguest for more information. INstruendum-Vitamin B1&B6-HPLC-UPLC-Calibrator Set FEN Pagina 12 van 12
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