Analysis of eight water soluble vitamins in ricebean (Vigna umbellata) varieties from NE India by reverse phase-HPLC

  • Rejaul Hoque Bepary
  • D. D. WadikarEmail author
  • P. E. Patki
Original Paper


A simple, quick, and sensitive HPLC procedure for simultaneous quantification of eight water-soluble vitamins (WSVs) in 11 ricebean (Vigna umbellata) varieties (JCR-08-7, JCR-08-8, JCR-08-10, JCR-08-12, JCR-08-15, JCR-08-16, JCR-08-32, JCR-13-11, JCR-13-13, JCR-50 and Nagadal) is described. The method includes enzymatic digestion followed by clean-up with solid phase extraction column and then analysis of extracted compounds by reverse phase column using UV-detector. The optimum conditions revealed that the method had an excellent linearity with good regression coefficient (r > 0.993), precision (0.3–2.77%), and accuracy (83–124%), and relatively short analysis time (13 min). The detection and quantification limits for all WSVs ranged 0.1470–1.1583 µg/mL and 0.4456–3.51 µg/mL, respectively. The average content (mg/100 g) of WSVs in 11 ricebean varieties was—B1: 0.579 ± 0.22, C: 2.111 ± 0.86, B3: 2.292 ± 1.15, B6 (pyridoxal-5-phosphate): 7.002 ± 1.31, B6 (pyridoxine hydrochloride): 2.79 ± 3.23, B5: 6.163 ± 3.35, B9: 0.289 ± 0.14, and B2: 0.163 ± 0.06. Principal components analysis showed that Nagadal belongs to third group which had the highest amount vitamin B1, vitamin B3, vitamin B5, and vitamin B2.


Water soluble vitamins HPLC Ricebean Method validation 



The authors acknowledge Ministry of Minority Affairs, Govt. of India, New Delhi and UGC, New Delhi for providing Fellowship to carry out the research works. The authors are gratefully acknowledge the Director, DFRL, Mysore for providing facilities, the support and encouragement to carry out this research and Dr (Mrs) Seuji. Borah Neog, Principal Scientist, AICRP on Forage Crops, Department of Plant Breeding and Genetics, Assam Agricultural University, Jorhat for providing the RB varieties for study.


  1. 1.
    P. Chen, R. Atkinson, W.R. Wolf, J. AOAC Int. 92, 680 (2009)Google Scholar
  2. 2.
    G.F.M. Ball, Vitamins in Foods: Analysis, Bioavailability, and Stability (CRC Press, Boca Raton, 2006), pp. 3–18Google Scholar
  3. 3.
    R. Rucker, J. Suttie, D. McCormick, L. Machlin, Handbook of Vitamins, 3rd edn. (Marcel Dekker, New York, 2001)Google Scholar
  4. 4.
    M. Benvenuti, E. Riches, The rapid analysis of 10 water-soluble vitamins, caffeine, and six common food dyes using acquity UPLC with UV detection, (Application note, Milford Waters Corporation, 2009). Accessed Dec 2017
  5. 5.
    C.J. Blake, Anal. Bioanal. Chem. 389, 63 (2007)Google Scholar
  6. 6.
    V. Spínolaa, E.J. Llorent-Martíneza, P.C. Castilhoa, J. Chromatogr. A 1369, 2 (2014)Google Scholar
  7. 7.
    B. Tiwari, N. Singh, Pulse Chemistry and Technology (The Royal Society of Chemistry, Cambridge, 2012) pp. 54Google Scholar
  8. 8.
    R.H. Bepary, D.D. Wadikar, P.E. Patki, Nutr. Food Sci. 46, 412 (2016)Google Scholar
  9. 9.
    R.H. Bepary, D.D. Wadikar, S.B. Neog, P.E. Patki, J. Food Sci. Technol. 54, 973 (2017)Google Scholar
  10. 10.
    D. Kaur, A.C. Kapoor, Food Chem. 43, 19 (1992)Google Scholar
  11. 11.
    M. Ciulu, S. Solinas, I. Floris, A. Panzanelli, M.I. Pilo, P.C. Piu, N. Spano, G. Sannaa, Talanta 83, 924 (2011)Google Scholar
  12. 12.
    J. Aslam, M.S. Mohajir, S.A. Khan, A.Q. Khan, Afr. J. Biotechnol. 7(14), 2310 (2008)Google Scholar
  13. 13.
    S. Rokayya, Y. Li, B. Qi, S. Wang, Q. Zhang, F. Han, Y. Ma, J. Jing, L. Jiang, J. Chem. (2014). Google Scholar
  14. 14.
    A. Lebiedzińska, M.L. Marszałł, J. Kuta, P. Szefer, J. Chromatogr. A 1173, 71–80 (2007)Google Scholar
  15. 15.
    O. Heudi, T. Kilinc, P. Fontannaz, J. Chromatogr. A 1070, 49–56 (2005)Google Scholar
  16. 16.
    R. Ekinci, C. Kadakal, Acta Chromatogr. A 15, 289 (2005)Google Scholar
  17. 17.
    S. Ndaw, M.Bergaentzlé,D. Aoudé-Werner, C. Hasselmann, Food Chem. 71, 129 (2000)Google Scholar
  18. 18.
    ICH (ICH) Harmonised Tripartite Guideline Validation of analytical procedure: Test and MethodologyQ2 (R1) (Current step 4 version, European Union, USA Japan: ICH Expert Working Group, 2005) Accessed Dec 2017
  19. 19.
    G.A. Shabir, J of Chromatogr, A 987, 57 (2003)Google Scholar
  20. 20.
    V. Gökmen, N. Kahraman, N. Demir, J. Acar, J. Chromatogr. A 881, 309–316 (2000)Google Scholar
  21. 21.
    S. Vidovic, B. Stojanovic, J. Veljkovi, L. Prăzić-Arsić, G. Roglić, D. Manojlović, J. Chromatogr. A 1202, 155–162 (2008)Google Scholar
  22. 22.
    R. Katock, J. Food Sci. 78, C8 (2013)Google Scholar
  23. 23.
    T. Longvah, R. Ananthan, K. Bhaskarachary, K. Venkaiah, Indian Food Composition Tables (ICMR, Department of Health Research Ministry of Health and Family Welfare, GOI, Jamai Osmania, 2017)Google Scholar
  24. 24.
    E. Azarpazhooh, J.I. Boye, Dry Bean and Pulses Production, Processing and Nutrition, 1st edn, (Wiley, New York, 2013)Google Scholar
  25. 25.
    D.C. Woollard, H.E. Indyk, S.K. Christiansen, Food Chem. 69, 201–208 (2000)Google Scholar
  26. 26.
    R. Engel PhD Thesis (submitted to Corvinus University of Budapest, 2009)Google Scholar
  27. 27.
    C. Pakin, M. Bergaentzlé, V. Hubscher, D. Aoudé-Werner, C. Hasselmann, J. Chromatogr. A 1035, 87–95 (2004)Google Scholar
  28. 28.
    A. Gonthier, V. Fayol, J. Viollet, D.J. Hartmann, Food Chem. 63, 287–294 (1998)Google Scholar
  29. 29.
    M. Vasic, J. Gvozdanovic-Varga, J. Cervenski, Genetika 40(1), 23–30 (2008)Google Scholar
  30. 30.
    I. Kabelová, M. Dvořáková, H. Cˇ Čížková, P. Dostálek, K. Melzoch, J. Food Compos. Anal. 21, 736–741 (2008)Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Rejaul Hoque Bepary
    • 1
  • D. D. Wadikar
    • 1
    Email author
  • P. E. Patki
    • 1
  1. 1.DRDO-Defence Food Research LaboratoryMysoreIndia

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