Separation and identification of folic acid and its related substances by liquid chromatography-mass spectrometry

NING Manru; XU Kehui; HANG Taijun; SONG Min

doi:10.11665/j.issn.1000-5048.2024052201

Journal of China Pharmaceutical University > 2025 > 56(1): 56-64. > DOI: 10.11665/j.issn.1000-5048.2024052201

NING Manru, XU Kehui, HANG Taijun, et al. Separation and identification of folic acid and its related substances by liquid chromatography-mass spectrometry[J]. J China Pharm Univ, 2025, 56(1): 56 − 64. DOI: 10.11665/j.issn.1000-5048.2024052201

Citation:

PDF (1053 KB)

Separation and identification of folic acid and its related substances by liquid chromatography-mass spectrometry

Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing 210009, China

More Information

Received Date: May 21, 2024

Graphical Abstract

Abstract

Abstract

The related substances in folic acid were separated on an ODS column by linear gradient elution using the mixture of 10 mmol/L ammonium acetate buffer solution and methanol (pH 6.3) as the mobile phase, and their accurate mass-charge ratio and elemental composition of parent and product ions were detected by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF/MS). The structures of the related substances were then determined by MS spectra elucidation, organic reaction mechanism analysis, or reference substances confirmation. Under the established LC-ESI-Q-TOF/MS conditions, folic acid and its related substances were adequately separated. Among the 23 main related substances found, 2 were known impurities listed in European Pharmacopoeia, 4 were consistent with the literature reports, and the remaining 17 were newly identified unknown related substances in this study. The results of separation and identification can provide some useful reference for the quality control of folic acid.
- folic acid,
- related substances,
- degradation products,
- structural identification,
- LC-MS

FullText(HTML)

References (16)

References

[1]	Lassi ZS, Salam RA, Haider BA, et al. Folic acid supplementation during pregnancy for maternal health and pregnancy outcomes[J]. Cochrane Database Syst Rev, 2013, 2013(3): CD006896.
[2]	Wiltshire E, Peña AS, MacKenzie K, et al. High dose folic acid is a potential treatment for pulmonary hypertension, including when associated with COVID-19 pneumonia[J]. Med Hypotheses, 2020, 143: 110142. doi: 10.1016/j.mehy.2020.110142
[3]	Goh YI, Koren G. Folic acid in pregnancy and fetal outcomes[J]. J Obstet Gynaecol, 2008, 28(1): 3-13. doi: 10.1080/01443610701814195
[4]	Feng Y, Wang S, Chen RS, et al. Maternal folic acid supplementation and the risk of congenital heart defects in offspring: a meta-analysis of epidemiological observational studies[J]. Sci Rep, 2015, 5: 8506. doi: 10.1038/srep08506
[5]	Wang WW, Wang XS, Zhang ZR, et al. A meta-analysis of folic acid in combination with anti-hypertension drugs in patients with hypertension and hyperhomocysteinemia[J]. Front Pharmacol, 2017, 8: 585. doi: 10.3389/fphar.2017.00585
[6]	Ting PC, Lee WR, Huo YN, et al. Folic acid inhibits colorectal cancer cell migration[J]. J Nutr Biochem, 2019, 63: 157-164. doi: 10.1016/j.jnutbio.2018.09.020
[7]	Qin XH, Cui YM, Shen L, et al. Folic acid supplementation and cancer risk: a meta-analysis of randomized controlled trials[J]. Int J Cancer, 2013, 133(5): 1033-1041. doi: 10.1002/ijc.28038
[8]	Delmonte L, Jukes TH. Folic acid antagonists in cancer chemotherapy[J]. Pharmacol Rev, 1962, 14: 91-135.
[9]	Chinese Pharmacopoeia Commission. Chinese Pharmacopoeia : part 4(中华人民共和国药典: 四部)[S]. Beijing: China Medical Science Press, 2020.
[10]	The United States Pharmacopeial 43-The National Formulary 38 : volume Ⅱ (美国药典43-国家处方集38: 卷2)[S]. Rockville: The United States Pharmacopeial Convention, 2020: 2680-2682.
[11]	European Pharmacopeia 10.0 : volume Ⅱ (欧洲药典10.0: 卷2)[S]. France: European Directorate for the Quality of Medicine&Healthcare, 2020: 3116-3117.
[12]	Shen QR, He Q, Pan YJ, et al. Detection, isolation, and characterization of a novel impurity from several folic acid products[J]. J Zhejiang Univ Sci B, 2022, 23(7): 597-606. doi: 10.1631/jzus.B2100970
[13]	Thomas AH, Lorente C, Capparelli AL, et al. Fluorescence of pterin, 6-formylpterin, 6-carboxypterin and folic acid in aqueous solution: pH effects[J]. Photochem Photobiol Sci, 2002, 1(6): 421-426. doi: 10.1039/b202114e
[14]	Thomas AH, Suárez G, Cabrerizo FM, et al. Study of the photolysis of folic acid and 6-formylpterin in acid aqueous solutions[J]. J Photochem Photobiol A Chem, 2000, 135(2/3): 147-154.
[15]	Cabrerizo F, Thomas A, Lorente C, et al. Generation of reactive oxygen species during the photolysis of 6-(hydroxymethyl)pterin in alkaline aqueous solutions[J]. Helv Chim Acta, 2004, 87(2): 349-365. doi: 10.1002/hlca.200490032
[16]	Off MK, Steindal AE, Porojnicu AC, et al. Ultraviolet photodegradation of folic acid[J]. J Photochem Photobiol B, 2005, 80(1): 47-55. doi: 10.1016/j.jphotobiol.2005.03.001