Food Analytical Methods

, Volume 10, Issue 7, pp 2444–2453 | Cite as

Development of a Simple, Fast, and Quantitative Lateral Flow Immunochromatographic Strip for Folic Acid

  • Zu-Pei Liang
  • Wu-Zu Ha
  • Zhi-Li Xiao
  • Hong-Tao Lei
  • Yu-Dong Shen
  • Yuan-Ming Sun
  • Hong Wang
  • Jin-Yi Yang
  • Zhen-Lin Xu


Folic acid (FA) is an important B vitamin naturally found in foods and also used in dietary supplements and food fortification. To monitor FA content is important to guarantee food quality and also to prevent its abuse in dietary supplements. In this work, we developed a simple and robust lateral flow immunochromatographic assay (ICA) for the routine screening of FA in food samples. FA was conjugated to carrier protein as immunogen and used to produce specific polyclonal antibody (pAb). The pAb was conjugated with gold nanoparticle and used to develop a lateral flow ICA. After the careful optimization of ICA conditions, the assay showed an IC50 of 51.8 ng mL−1 and linear range from 23.4 to 114.5 ng mL−1 for FA in assay buffer, with the assist of a DY6510 strip reader within 8 min. Using milk powder as a matrix model, after a 15-fold dilution to eliminate matrix effect, the ICA showed intra-assay recoveries from 94.2 to 107.0% with coefficient of variance (CV) below 15% and inter-assay recoveries from 91.4% to 111.3% with CV below 18%. Good agreement was obtained between the results of ICA and ciELISA and standard HPLC. It is an ideal screening tool for the fast monitoring of FA in a simple, fast, and low-cost manner.


Folic acid Polyclonal antibody Immunochromatographic assay Quantitative detection 


Compliance with Ethical Standards


This work was supported by the Guangdong Natural Science Funds for Distinguished Young Scholar (2014A030306026), Guangdong Special Support Program (2014TQ01N109), and Guangdong Young Teachers Program on Higher Education (Y920/4026).

Conflict of Interest

Zu-Pei Liang declares that he has no conflict of interest. Wu-Zu Ha declares that he has no conflict of interest. Zhi-Li Xiao declares that he has no conflict of interest. Hong-Tao Lei declares that he has no conflict of interest. Yu-Dong Shen declares that he has no conflict of interest. Yuan-Ming Sun declares that he has no conflict of interest. Hong Wang declares that he has no conflict of interest. Jin-Yi Yang declares that she has no conflict of interest. Zhen-Lin Xu declares that she has no conflict of interest.

Ethical Approval

All institutional and national guidelines for the care and use of laboratory animals were followed.

Informed Consent

Not applicable

Supplementary material

12161_2017_804_MOESM1_ESM.doc (8.9 mb)
ESM 1 (DOC 9100 kb)


  1. Anonymous (2005) Human energy requirements: report of a joint FAO/WHO/UNU Expert Consultation. Food Nutr Bull 26(1):166CrossRefGoogle Scholar
  2. Belz S, Nau H (1998) Determination of folate patterns in mouse plasma, erythrocytes, and embryos by HPLC coupled with a microbiological assay. Anal Biochem 265(1):157–166CrossRefGoogle Scholar
  3. Breshkovskaya OET, Elena VGM, Paredes LO (2010) Determination of the folate content in cladodes of nopal (Opuntia Ficus Indica) by microbiological assay utilizing Lactobacillus casei (ATCC 7469) and enzyme-linked lmmunosorbent assay. J. Agr Food Chem 58(10):6472–6475CrossRefGoogle Scholar
  4. Christodoulides N, Mohanty S, Miller CS, Langub MC, Floriano PN, Dharshan P, Ali MF, Bernard B, Romanovicz D, Anslyn E, Fox PC, Mcdevitt JT (2005) Application of microchip assay system for the measurement of C-reactive protein in human saliva. Lab Chip 5(3):261–269CrossRefGoogle Scholar
  5. Chun J, Martin JA, Chen LW, Lee J, Ye L, Eitenmiller RR (2006) A differential assay of folic acid and total folate in foods containing enriched cereal-grain products to calculate mu g dietary folate equivalents (mu g DFE). J Food Compos Anal 19(2–3):182–187CrossRefGoogle Scholar
  6. Dai J, Xu Z, Liu F, Yang J, Sun Y, Wang H, Lei H, Shen Y (2015) Determination of furaltadone metabolite in fish by chemiluminescence enzyme immunoassay. Chinese J Anal Chem 43(6):871–875CrossRefGoogle Scholar
  7. De Brouwer V, Storozhenko S, Van De Steene JC, Wille SM (2008) Optimisation and validation of a liquid chromatography–tandem mass spectrometry method for folates in rice. J Chromatogr A 1215:126–132CrossRefGoogle Scholar
  8. Duan H, Chen X, Jiang H, Shen J, Dong S, Xiong Y, Wang A (2015) Quantum-dot submicrobead-based immunochromatographic strip for rapid and quantitative detection of Plasmodium falciparum. Chinese J. Anal Chem 43(3):338–343Google Scholar
  9. Han JY, Tyler RT (2003) Determination of folate concentrations in pulses by a microbiological method employing trienzyme extraction. J. Agr Food Chem 51(18):5315–5318CrossRefGoogle Scholar
  10. Hinterberger M, Fischer P (2013) Folate and Alzheimer: when time matters. J Neural Transm 120(1SI):211–224CrossRefGoogle Scholar
  11. Hou W, Wang S, Wang X, Han X, Fan H, Cao S, Yue J, Wang Q, Jiang W, Ding C, Yu S (2015) Development of colloidal gold immunochromatographic strips for detection of Riemerella anatipestifer. PLoS One 10:e01229523Google Scholar
  12. Iyer R, Tomar SK (2009) Folate: a functional food constituent. J Food Sci 74(9):R114–R122CrossRefGoogle Scholar
  13. Jastrebova J, Witthoft C, Grahn A, Svensson U, Jagerstad M (2003) HPLC determination of folates in raw and processed beetroots. Food Chem 80:579–588CrossRefGoogle Scholar
  14. Kotsopoulos J, Kim Y, Narod SA (2012) Folate and breast cancer: what about high-risk women? Cancer Cause Control 23(9):1405–1420CrossRefGoogle Scholar
  15. Krista SC, Lynn BB, Robert JB (2011) Folic acid food fortification—its history, effect, concerns, and future directions. Nutrients 3:370–384CrossRefGoogle Scholar
  16. Lermo A, Fabiano S, Hernandez S, Galve R, Marco MP, Alegret S, Pividori MI (2009) Immunoassay for folic acid detection in vitamin-fortified milk based on electrochemical magneto sensors. Biosens Bioelectron 24(7):2057–2063CrossRefGoogle Scholar
  17. Lubecka PK, Kaufman SA, Stefanska B, Fabianowska MK (2013) Folic acid enforces DNA methylation-mediated transcriptional silencing of PTEN, APC and RARbeta2 tumour suppressor genes in breast cancer. Biochem Bioph Res Co 430(2):623–628CrossRefGoogle Scholar
  18. Lucock M, Yates Z (2005) Folic acid—vitamin and panacea or genetic time bomb? Nat Rev Genet 6(3):235–240CrossRefGoogle Scholar
  19. Luo L, Xu ZL, Yang JY, Xiao ZL, Li Y, Beier RC, Sun YM, Lei HT, Wang H, Shen YD (2014) Synthesis of novel haptens and development of an enzyme-linked immunosorbent assay for quantification of histamine in foods. J. Agr Food Chem 62(51):12299–12308CrossRefGoogle Scholar
  20. Luo L, Yang JY, Xiao ZL, Zeng DP, Li Y, Shen YD, Sun YM, Lei HT, Wang H, Xu ZL (2015) A sensitivity-enhanced heterologous immunochromatographic assay based on a monoclonal antibody for the rapid detection of histamine in saury samples. RSC Adv 5(96):78833–78840CrossRefGoogle Scholar
  21. Malin BC, Johan L (2000) Biosensor-based determination of folic acid in fortified food. Food Chem 70:523–532CrossRefGoogle Scholar
  22. Maria V, Chandra H, Martin PB (2013) Folic acid-fortified flour: optimised and fast sample preparation coupled with a validated high-speed mass spectrometry analysis suitable for a fortification monitoring program. Food Anal Methods 6:1416–1423CrossRefGoogle Scholar
  23. Nasr HC, Jalloul M, Sabbah M, Adib SM (2012) Awareness and intake of folic acid for the prevention of neural tube defects among Lebanese women of childbearing age. Matern Child Health J 16(1):258–265CrossRefGoogle Scholar
  24. Nguyen MT, Indrawati HM (2003) Model studies on the stability of folic acid and 5-methyltetrahydrofolic acid degradation during thermal treatment in combination with high hydrostatic pressure. J. Agr Food Chem 51(11):3352–3357CrossRefGoogle Scholar
  25. Pérez P, Cancho G, García F, Simal G (2006) Screening for folic acid content in vitamin-fortiWed beverages. Food Control 17:900–904CrossRefGoogle Scholar
  26. Philip D (2008) Synthesis and spectroscopic characterization of gold nanoparticles. Spectrochim Acta A 71(1):80–85CrossRefGoogle Scholar
  27. Quan Y, Zhang Y, Wang S, Lee N, Kennedy IR (2006) A rapid and sensitive chemiluminescence enzyme-linked immunosorbent assay for the determination of fumonisin B-1 in food samples. Anal Chim Acta 580(1):1–8CrossRefGoogle Scholar
  28. Robert JP, Hertrampf E, Vincent PF, Paul MT (2003) Mass spectral determinations of the folic acid content of fortified breads from Chile. J Food Compos Anal 16:281–286CrossRefGoogle Scholar
  29. Shen J, Zhang Z, Yao Y, Shi W, Liu Y, Zhang S (2006) A monoclonal antibody-based time-resolved fluoroimmunoassay for chloramphenicol in shrimp and chicken muscle. Anal Chim Acta 575(2):262–266CrossRefGoogle Scholar
  30. Suryoprabowo S, Liu L, Peng J, Kuang H, Xu C (2014) Development of a broad specific monoclonal antibody for fluoroquinolone analysis. Food Anal Method 7(10):2163–2168CrossRefGoogle Scholar
  31. Sweeney MR, Mcpartlin J, Scott J (2007) Folic acid fortification and public health: report on threshold doses above which unmetabolised folic acid appear in serum. BMC Public Health 7(41)Google Scholar
  32. Wang X, Li K, Shi D, Xiong N, Jin X, Yi J, Bi D (2007) Development of an immunochromatographic lateral-flow test strip for rapid detection of sulfonamides in eggs and chicken muscles. J Agr Food Chem 55(6):2072–2078CrossRefGoogle Scholar
  33. Wright AJA, Dainty JR, Finglas PM (2007) Folic acid metabolism in human subjects revisited: potential implications for proposed mandatory folic acid fortification in the UK. Brit J Nutr 98(4):667–675CrossRefGoogle Scholar
  34. Zhang B, Zhao L, Lin J (2008) Determination of folic acid by chemiluminescence based on peroxomonosulfate-cobalt(II) system. Talanta 74(5):1154–1159CrossRefGoogle Scholar
  35. Zhang T, Xue H, Zhang B, Zhang Y, Song P, Tian X, Xing Y, Wang P, Meng M, Xi R (2012) Determination of folic acid in milk, milk powder and energy drink by an indirect immunoassay. J Sci Food Agr 92(11):2297–2304CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Zu-Pei Liang
    • 1
    • 2
  • Wu-Zu Ha
    • 3
  • Zhi-Li Xiao
    • 1
  • Hong-Tao Lei
    • 1
  • Yu-Dong Shen
    • 1
  • Yuan-Ming Sun
    • 1
  • Hong Wang
    • 1
  • Jin-Yi Yang
    • 1
  • Zhen-Lin Xu
    • 1
  1. 1.Guangdong Provincial Key Laboratory of Food Quality and SafetySouth China Agricultural UniversityGuangzhouChina
  2. 2.Guangdong Testing Institute of Product Quality SupervisionGuangzhouChina
  3. 3.College of Chemistry and Chemical EngineeringWuhan Textile UniversityWuhanChina

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