Liquid Chromatography Method for the Simultaneous Quantification of Biotin and Vitamin B12 in Vitamin B Supplements

Abstract

The determination of cyanocobalamin (vitamin B12) and biotin has always been challenging because of the lack of a chromophore for biotin and trace level input for vitamin B12 in supplements. Microbiological assay methods are currently used for quantitation. However, these methods are time consuming, may lack specificity, and have high imprecision. Our laboratory developed and validated an LC method for the simultaneous quantitation of vitamin B12 and biotin. This LC method uses a single quadruple mass analyzer to detect biotin and vitamin B12 at 245.10 and 678.29, respectively. The mass analyzer allows for low limits of quantitation (biotin: 1 ng/mL; vitamin B12: 4 ng/mL). Precision results showed that injections are repeatable without the use of an internal standard (RSD< 5%). Single analyst ( = 5: RSD < 3%), within lab ( = 10: RSD < 8%), and multilab ( = 20: RSD < 13%) precision results were also much better than those reported by microbiological assay methods. Linearity was evaluated between 92.00 ng/mL and 9200 ng/mL (² 09916) for biotin and between 4.846 ng/mL and 484.6 ng/mL (² 0.9999) for vitamin B12. The method is accurate between 20 ng/mL and 60 ng/mL for vitamin B12 and between 400 ng/mL and 1200 ng/mL for biotin. The results show a simple, accurate, and precise method for the quantitation of vitamin B12 and biotin. This work demonstrates that single quadrupole mass analyzers can be successfully used to quantify trace level analytes in quality control laboratories.

Immunological strip sensor for the rapid determination of niacin in dietary supplements and foods

“…Furthermore, the fact that niacin and nicotinamide are also nutrients for the growth of Lactobacillus plantarum makes the assay inaccurate and unable to differentiate between niacin and nicotinamide. [11][12][13] For analysis using specialized instruments, high performance liquid chromatography (HPLC), gas chromatography (GC), liquid chromatography-mass spectrometry (L-MS), and gas chromatography-mass spectrometry (GC-MS) have been used for the detection of niacin. 14,15 Instruments using chemical analysis are well established and precise, making the results highly sensitive and reliable.…”

The Fast Quantification of Vitamin B12 in Milk Powder by High-Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry

“…Currently, the accepted methods for determining vitamin B12 in healthcare supplies are high-performance liquid chromatography (HPLC) [9][10][11][12] and the microbiological method, which is only used for infant food and dairy products [13,14]. Nevertheless, because of the low sensitivity of the HPLC method, it was necessary to enrich and purify it using solid-phase extraction to achieve a higher method detection limit [15][16][17][18]. However, this entire process is intricate and challenging to control.…”

Determination of Vitamin B12 in Milk and Dairy Products by Isotope-Dilution Liquid Chromatography Tandem Mass Spectrometry

“…e existing reported methods for determination of VB 12 in infant formula mainly include microbiological assays [9,10], high-performance liquid chromatography (HPLC) [11][12][13][14][15][16][17][18][19], liquid chromatography-tandem mass spectrometry (LC-MS/MS) [20][21][22][23][24][25][26], and liquid chromatography and inductively coupled plasma-mass spectrometry (LC-ICP-MS) [27][28][29]. e microbiological assays usually have a long measurement time and complicated operation.…”