Determination of cyanocobalamin by isotope dilution LC-MS/MS

D’Ulivo, Lucia; Yang, Lu; Ding, Jianfu; Leek, Donald M.; Thibeault, Marie-Pier; Mester, Zoltán

doi:10.1016/j.aca.2017.07.065

Cited by 17 publications

(12 citation statements)

References 27 publications

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“…The extraction and purification were developed based on the published native SIDA method [ 7 ] as well as previous cyanidation conversion-based methods [ 12 – 14 , 19 ]. During method development, various sample preparation methods were tested on a pork fillet sample in order to investigate and optimize the conversion process.…”

Section: Methodsmentioning

confidence: 99%

Challenges in the determination of total vitamin B12 by cyanidation conversion: insights from stable isotope dilution assays

et al. 2023

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Previous methods for vitamin B12 (B12) analysis have extensively used cyanidation conversion with the intention of converting all cobalamins to cyanocobalamin (CNCbl) for total B12 determination. This approach has been favored for its advantages in reducing the number of analytes, increasing analyte concentration, and improving analyte stability. However, the present study revealed underlying limitations associated with this approach. First, a stable isotope dilution assay (SIDA) determining total B12 as CNCbl after cyanidation conversion (conversion SIDA method) was developed. Method validation demonstrated good sensitivity, recovery, accuracy, and reproducibility for the target analyte CNCbl. However, subsequent application of the conversion method to real meat samples showed incomplete conversions of cobalamins. These inconsistencies revealed day-to-day variability and reliability challenges associated with the cyanidation process. It was not possible to identify this issue during method validation as CNCbl was spiked as the sole analyte and it requires no further cyanidation conversion. The application of LC-MS/MS enabled the detection of trace amounts of unconverted cobalamins. Nevertheless, this approach remains restricted by instrument sensitivity and stability as well as the performance of immunoaffinity purification for different vitamers. Further development of a reliable monitoring method is a prerequisite for further optimization of the cyanidation process. However, significant improvements of analytical instrumentation in terms of sensitivity and stability are required to overcome the current limitations. Graphical abstract

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Section: Methodsmentioning

confidence: 99%

Challenges in the determination of total vitamin B12 by cyanidation conversion: insights from stable isotope dilution assays

et al. 2023

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“…Subsequent method development and validation by Matsumoto et al analyzed SRM 3280 along with samples representative of multivitamin DS with or without elements and other NP ingredients like coenzyme Q10 to assess the accuracy and uncertainty range of their HPLC-UV detection method ( 57 ). D'Ulivo et al developed a novel ID method for cyanocobalamin using an isotopically enriched 13 C 15 N cyanocobalamin as the internal standard and validated the LC-MS/MS method using SRM 3280 ( 70 ). The validated ID-LC-MS/MS method was then employed to certify the content of cyanocobalamin in two multivitamin CRMs recently issued by the National Research Center of Canada (NRCC), i.e., VITA-1 and VITB-1.…”

Section: Case Studies Of Research Using Dietary Supplement Matrix Crmsmentioning

confidence: 99%

The Importance of Reference Materials and Method Validation for Advancing Research on the Health Effects of Dietary Supplements and Other Natural Products

2021

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Insufficient assessment of the identity and chemical composition of complex natural products, including botanicals, herbal remedies, and dietary supplements, hinders reproducible research and limits understanding mechanism(s) of action and health outcomes, which in turn impede improvements in clinical practice and advances in public health. This review describes available analytical resources and good methodological practices that support natural product characterization and strengthen the knowledge gained for designing and interpreting safety and efficacy investigations. The practice of validating analytical methods demonstrates that measurements of constituents of interest are reproducible and appropriate for the sample (e.g., plant material, phytochemical extract, and biological specimen). In particular, the utilization of matrix-based reference materials enables researchers to assess the accuracy, precision, and sensitivity of analytical measurements of natural product constituents, including dietary ingredients and their metabolites. Select case studies are presented where the careful application of these resources and practices has enhanced experimental rigor and benefited research on dietary supplement health effects.

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“…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.…”

Section: Introductionmentioning

confidence: 99%

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

Huang

Zhang

Cai

2022

Journal of Food Quality

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An isotope-dilution liquid chromatography tandem mass spectrometry method was established for the determination of vitamin B12 in milk and dairy products. The samples were spiked with stable isotope-labeled vitamin B12 and digested by pepsin and amylase. The various forms of cobalamin were transformed to cyanocobalamin by potassium cyanide after they were released from the enzymatically digested samples. Cyanocobalamin was extracted and purified by an immunoaffinity SPE cartridge and then measured in multiple reaction monitoring mode (MRM). The linear correlation coefficient (R2) of this method was greater than 0.999 in the range of 2–100 ng/mL. The detection limit and the quantification limit were 0.5 μg/kg and 1.0 μg/kg, respectively. The spiked recoveries ranged from 92.0% to 99.4% at the three spiked levels with the relative standard deviation (RSD) between 1.89% and 4.51%. The measured results of NIST SRM1849a and NIST SRM1869a by the current method are all within the reference value range. The Z value was 0.8 during participating in the FAPAS proficiency test using the developed method in 2021. The method is simple, rapid, accurate, and sensitive, and it is suitable for the determination of vitamin B12 in different types of milk and dairy products such as whey powder, whole milk powder, pure milk, fermented milk, infant formula, and prescription food for special medical purposes.

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Determination of cyanocobalamin by isotope dilution LC-MS/MS

Cited by 17 publications

References 27 publications

Challenges in the determination of total vitamin B12 by cyanidation conversion: insights from stable isotope dilution assays

Challenges in the determination of total vitamin B12 by cyanidation conversion: insights from stable isotope dilution assays

The Importance of Reference Materials and Method Validation for Advancing Research on the Health Effects of Dietary Supplements and Other Natural Products

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

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