
Primesep C separates B vitamins with baseline resolution by a combination of cation exchange, complex formation, and hydrophobic interactions. Vitamin B1 (thiamine), vitamin B2 (riboflavin), and vitamin B6 (pyridoxine) are separated with a mobile phase of water, acetonitrile (MeCN, ACN) and triethylamine (TEA) phosphate with UV detection at 280 nm.
Vitamin B1 (Thiamine) Vitamin B2 (Riboflavin) Vitamin B3 (Niacin) Riboflavin (Vitamin B2) Thiamine (Vitamin B1) Pyridoxine Vitamins
UV Detection

Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Both columns allow peak order reversal by simply changing the acetonitrile concentration in the mobile phase. Vitamin B2 is especially sensitive to acetonitrile concentration. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either trifluoroacetic acid (TFA) or acetic acid (HOAc) with UV detection at 250 nm.
Vitamin C Vitamin B2 (Riboflavin)
UV Detection

Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Peak order reversal is exhibited on these two columns with the same mobile phases due to their different polarity. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either formic or acetic acid (HOAc) with UV detection at 250 nm.
Vitamin B2 (Riboflavin) Riboflavin (Vitamin B2) Vitamin C Ascorbic Acid
UV Detection

Vitamin C (ascorbic acid) and Vitamin B2 (riboflavin) are water-soluble vitamins that are not stored in the body and must be replaced each day. Primesep D easily resolves Vitamins C and B2 and the elution order can be reversed by adjusting acetonitrile and TFA concentrations in the mobile phase. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoroacetic acid (TFA) and UV detection at 250 nm.
Vitamin B2 (Riboflavin) Vitamin C
UV Detection
 Separation of Vitamin B2 and Vitamin C is demonstrated on a Primesep D mixed-mode column. This column can be used for analysis of other hydrophobic or neutral vitamins and supplements.
Vitamin B2 (Riboflavin) Vitamin C
UV Detection

Vitamin C (ascorbic acid) and Vitamins Group B are separated on Obelisc N mixed-mode column. Method can be used in quantitation and determination of polar vitamins in various formulations and dietary supplements. HPLC method can be based on UV, Evaporative Light Scattering Detection (ESLD), RI or MS detection. Effect of sample matrix can be eliminated by changing mobile phase conditions. Buffer concentration, buffer pH and amount of ACN will affect every vitamin differently due to difference in polar and ionic properties.
Pyridoxine Vitamin B6 (Pyridoxine) 4-Aminopyridine/Fampridine Vitamin B2 (Riboflavin) 3-Aminopyridine Isonicotinic Acid Nicotinic Acid Barbituric Acid Ascorbic Acid
UV Detection

Separation of vitamins group B is achieved on Obelisc N column in HILIC mixed-mode. Vitamins of this group are different in polarity and ionic properties. Retention and separation is achieved by optimization of amount of ACN, buffer and buffer pH. Combination of UV and ELSD detection is used to monitor HPLC separation. B vitamins are water-soluble vitamins that play an important role in cell metabolism. Supplements containing all six are generally referred to as a vitamin B complex. Individual B vitamin supplements are referred to by the specific name of each vitamin. This method can be used to analyze individual B vitamins as well as vitamin B complex. Isolation of impurities as well as degradation products is possible by preparative chromatography.
Vitamin B3 (Niacin) Vitamin B6 (Pyridoxine) Vitamin B2 (Riboflavin) Vitamin B5 (Pantothenic Acid) Vitamin B1 (Thiamine) Nicotinic Acid
UV Detection ELSD/MS Detection
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