Plant Foods for Human Nutrition

, Volume 73, Issue 3, pp 166–171 | Cite as

Methylmalonic Acid Levels and their Relation with Cobalamin Supplementation in Spanish Vegetarians

  • Angélica Gallego-Narbón
  • Belén Zapatera
  • Inmaculada Álvarez
  • M. Pilar VaqueroEmail author
Original Paper


Cobalamin deficiency represents a health issue for vegetarians, especially vegans, if supplements are not consumed. Vitamin B12 serum levels, traditionally used to assess the vitamin B12 status, can be normal under functional deficiency conditions. In this regard, methylmalonic acid (MMA) has proven to be a more specific marker to detect subclinical vitamin B12 deficiency. In this study, we present for the first time the cobalamin status of Spanish vegetarians using both vitamin B12 and MMA markers, and the effects of the plant-based diet and the intake of vitamin B12 supplements. Healthy adults were recruited (n = 103, 52% vegans). Dietary preferences and use of supplements were assessed by questionnaires and serum samples were collected and stored. Vitamin B12 was measured by chemiluminiscence and MMA by liquid chromatography tandem mass spectrometry (LC-MS/MS) using solid phase extraction for sample preparation. Obtained values, median (IQR), were: vitamin B12, 278.9 (160.2) pmol/l and MMA, 140.2 (78.9) nmol/l. No significant differences between lacto-ovo vegetarians and vegans were observed. Considering these two markers, 10% of the participants were mild vitamin B12 deficient. Supplementation (75% of the participants) was associated with higher vitamin B12 (p < 0.001) and lower MMA (p = 0.012). In conclusion, Spanish vegetarians have low risk of vitamin B12 deficiency due to vitamin B12 supplementation and the MMA determination is useful to detect mild deficiency.


Methylmalonic acid Vitamin B12 Plant-based diet Vegetarians Vegans LC-MS/MS 



Body Mass Index


High concentration quality control


Low concentration quality control


Lacto-ovo vegetarians


Methylmalonic acid


Multiple reaction monitoring


Solid phase extraction


Liquid chromatography tandem mass spectrometry



The authors are grateful to Marta Novoa for her help in obtaining and storing serum samples. Ana Salvador is acknowledged for critical review of the manuscript. The study was supported by a research project financed by Zamdeh Laboratories (Veggunn study). A. Gallego-Narbón was funded by the Youth Employment Initiative (YEI) from the European Social Fund (ESF).

Compliance with Ethical Standards

Conflicts of Interest

The authors declare the absence of conflicts of interest. The human study was conducted according to the guidelines laid down in the Declaration of Helsinki and was approved by the Ethics Committees of Hospital Puerta de Hierro (Majadahonda, Madrid) and CSIC.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Metabolism and NutritionInstitute of Food Science, Technology and Nutrition (ICTAN-CSIC)MadridSpain
  2. 2.Unit Service of Analytical Techniques, Instrumentation and MicrobiologyInstitute of Food Science, Technology and Nutrition (ICTAN-CSIC)MadridSpain

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