Determination of the Chemical Stability of Cyanocobalamin in Medical Food by a Validated Immunoaffinity Column-Linked HPLC Method

Abstract

Cyanocobalamin, which plays an essential role in the body, is a synthetic form used in medical food. This present study aimed to develop an HPLC analysis method for determination cyanocobalamin and investigate the stability of cyanocobalamin in medical food. Validation of the developed method for cyanocobalamin was evaluated with linearity, LOD, LOQ, and accuracy. The linearity of this method was calculated with a value of the coefficient of determination (R2) ≥ 0.999. LOD and LOQ were 0.165 and 0.499 μg/kg, respectively. The recovery of medical food matrixes for accuracy was more than 97.63%. The validated method was applied for determining cyanocobalamin from medical foods. The developed method was used to examine the additives for cyanocobalamin protection. Ferric chloride and sorbitol alleviated cyanocobalamin degradation from heat and ascorbic acid. Especially, sorbitol showed a superior protective effect during the medical food production process. Therefore, this study suggests that sorbitol is a sweetener additive that prevents cyanocobalamin degradation by heat and the food matrix in medical food processing.

Vitamin B12 in Foods, Food Supplements, and Medicines—A Review of Its Role and Properties with a Focus on Its Stability

“…Its presence in solution is reported to cause up to 65% CNCbl degradation after 24 h of storage at 25 • C in the dark [122]. The destabilizing effect of ascorbic acid increases with the temperature, ascorbic acid concentration, and exposure of the solutions to light [122,179,181]. It also depends on the pH of the solution (faster reaction up to pH around 5.0 for both CNCbl and OHCbl) and on the vitamin B 12 form (MeCbl is the least stable vitamin B 12 form, followed by OHCbl and finally CNCbl) [122,145,179].…”

“…Therefore, Lee et al evaluated its adequacy as a CNCbl stabilizing agent during the production process of medical food. In addition to its protective effect on the thermal and ascorbic acid-induced degradation of CNCbl in solutions, Lee et al also demonstrated that sorbitol efficiently protected CNCbl against degradation in medical foods, caused by heat (two-step sterilization) and the food matrix, which also contained ascorbic acid [181].…”

Effects of metal ions on cyanocobalamin stability in heated milk protein‐based matrices

“…Factors capable of promoting B12 loss and thereby matrix variability include (in addition to the heat, oxygen, and B12 molecular form mentioned above) light, pH, reducing agents (including ascorbic acid, thiols, polyphenols, bisulfite, reducing sugars, ferrous salts), oxidising agents (including reactive oxygen species), other vitamins and/or decomposition products thereof, and sucrose (Hutchins et al ., 1956; Dolphin et al ., 1963; Ichikawa et al ., 2005; Ramasamy et al ., 2012; Ahmad et al ., 2014; Monajjemzadeh et al ., 2014; Salnikov et al ., 2014; Johns et al ., 2015; Dereven'kov et al ., 2016; Nakos et al ., 2017; Rempel et al ., 2018; Lie et al ., 2019; Schnellbaecher et al ., 2019; Salnikov et al ., 2021; Lee et al ., 2022).…”

“…Since infant formulas and other fortified foods may be formulated with vitamins (including CN‐B12) and minerals (including salts of the redox active metals iron and copper, and the redox inactive metal zinc), knowledge of fortified metal ion effects on CN‐B12 stability (particularly during processing) could facilitate strategies for optimising nutrient delivery by these fortified foods. In this connection, prior research has found that Fe 2+ may destabilise B12, and that the ascorbic acid‐induced decomposition of B12 may be substantially accelerated by Cu 2+ , but significantly inhibited by Fe 3+ (Rosenberg, 1956; Nakos et al ., 2017; Schnellbaecher et al ., 2019; Lee et al ., 2022). Accordingly, the objective of the present study was to evaluate the effects of redox active metal cations (Cu 2+ , Fe 2+ , Fe 3+ , Mn 2+ , Ce 3+ , and Cr 6+ ) and a redox inactive metal cation (Zn 2+ ) on the stability of CN‐B12 added to milk‐based matrices and subjected to heat.…”

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.…”