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Key Documents

G005625

HPLC Analysis of Water-Soluble B-Vitamins on Ascentis® Express C18

application for HPLC

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About This Item

technique(s)

HPLC: suitable

test parameters

column: Ascentis Express C18, 5 cm x 3.0 mm I.D., 2.7 μm particles (53811-U)
mobile phase: [A] 20 mM potassium phosphate, dibasic, pH 7; [B] methanol
gradient: 0.5% B for 1.3 min, 0.5-30%B over 1.7 min; to 30% B in 1.4 min, 2 min equilibration at 0.5% B
flow rate: 1.0 mL/min
pressure: 4130 psi (285 bar)
column temp.: 35 °C
detector: UV, 210 nm
sample: 9 vitamins in water, 10 μg/mL for nicotinic acid and folic acid, 12 μg/mL for thiamine hydrochloride and cyanocobalamin, (20 μg/mL for pyridoxine, 30 μg/mL for riboflavin, 40 μg/mL for niacineamide, 100 μg/mL for calcium pantothenate, 120 μg/mL for biotin)

suitability

application for HPLC

application(s)

vitamins, nutraceuticals, and natural products

Analysis Note

A mix of 9 water-soluble B-vitamins was separated on Ascentis Express C18 column

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany

Analyte

Description

    Nicotinic acid, ≥99.5% (HPLC)
    Calcium pantothenate, meets USP testing specifications, monograph mol wt. 476.53 (C18H32CaN2O10)
    Pyridoxine hydrochloride, ≥98% (HPLC)
    Nicotinamide, ≥98.5% (HPLC)
    Thiamine hydrochloride, reagent grade, ≥99% (HPLC)
    Biotin, ≥99.0% (T)
    Folic acid, analytical standard
    Cyanocobalamin (B12), analytical standard
    Riboflavin (B2), analytical standard
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