A Dual-Standard Reversed-Phase HPLC-UV Method for the Simultaneous Identification, Assay, and Impurities Analysis of Phenylboronic Acid: Using 4-Methoxyphenyl Boronic Acid as a Model Compound
Boronic acids (BAs) are versatile building blocks in organic synthesis and the pharmaceutical industry. A dual-standard reverse phase (RP) HPLC-UV method was developed for the simultaneous identification (as monomer), assay (as monomer), and impurities analysis of phenylboronic acid (PBA), using 4-methoxyphenyl boronic acid (4-MPBA) as a model compound. The method utilizes an Ascentis? Express 90 ? ES-Cyano column with 0.025% TFA in both mobile phases (A: water, B: methanol). It employs a simple gradient with an injection volume of 10 μL and a flow rate of 0.8 mL/min. This QC-friendly, single-sequence method achieves baseline separation of the analyte and its impurities. By utilizing a stable, readily available bromo-analog surrogate for quantitation, it bypasses the need for a single-component primary BA reference standard.
References
[1]
Fyfe, J.W.B. and Watson, A.J.B. (2017) Recent Developments in Organoboron Chemistry: Old Dogs, New Tricks. Chem, 3, 31-55. https://doi.org/10.1016/j.chempr.2017.05.008
[2]
Miyaura, N., Yamada, K. and Suzuki, A. (1979) A New Stereospecific Cross-Coupling by the Palladium-Catalyzed Reaction of 1-Alkenylboranes with 1-Alkenyl or 1-Alkynyl Halides. TetrahedronLetters, 20, 3437-3440. https://doi.org/10.1016/s0040-4039(01)95429-2
[3]
António, J.P.M., Russo, R., Carvalho, C.P., Cal, P.M.S.D. and Gois, P.M.P. (2019) Boronic Acids as Building Blocks for the Construction of Therapeutically Useful Bioconjugates. ChemicalSocietyReviews, 48, 3513-3536. https://doi.org/10.1039/c9cs00184k
[4]
Coghi, P.S., Zhu, Y., Xie, H., Hosmane, N.S. and Zhang, Y. (2021) Organoboron Compounds: Effective Antibacterial and Antiparasitic Agents. Molecules, 26, Article 3309. https://doi.org/10.3390/molecules26113309
[5]
Das, B.C., Nandwana, N.K., Das, S., Nandwana, V., Shareef, M.A., Das, Y., et al. (2022) Boron Chemicals in Drug Discovery and Development: Synthesis and Medicinal Perspective. Molecules, 27, Article 2615. https://doi.org/10.3390/molecules27092615
[6]
Adams, J. and Kauffman, M. (2004) Development of the Proteasome Inhibitor Velcade？ (Bortezomib). CancerInvestigation, 22, 304-311. https://doi.org/10.1081/cnv-120030218
[7]
U.S. Food and Drug Administration (2015) Velcade (Bortezomib) Injection. Application No. 021602. https://www.fda.gov/media/94379/download
[8]
U.S. Food and Drug Administration (2019) Efficacy and Safety Evaluation of AN2690 Topical Solution to Treat Onychomycosis of the Toenail. Clinical Trial No. NCT01270971. https://clinicaltrials.gov/study/NCT01270971?term=NCT01270971&rank=1
[9]
U.S. Food and Drug Administration (2015) Millennium Pharmaceuticals Inc., NINLARO (Ixazomib) Capsules. Application No. 208462. https://www.accessdata.fda.gov/drugsatfda_docs/nda/2015/208462Orig1s000Approv.pdf
[10]
U.S. Food and Drug Administration (2016) Eucrisa (Crisaborole) Ointment. Application No. 207695. https://www.accessdata.fda.gov/drugsatfda_docs/nda/2016/207695Orig1s000Lbl.pdf
[11]
Maslah, H., Skarbek, C., Pethe, S. and Labruère, R. (2020) Anticancer Boron-Containing Prodrugs Responsive to Oxidative Stress from the Tumor Microenvironment. EuropeanJournalofMedicinalChemistry, 207, Article ID: 112670. https://doi.org/10.1016/j.ejmech.2020.112670
[12]
Faraco, M., Fico, D., Pennetta, A. and De Benedetto, G.E. (2016) New Evidences on Efficacy of Boronic Acid-Based Derivatization Method to Identify Sugars in Plant Material by Gas Chromatography–mass Spectrometry. Talanta, 159, 40-46. https://doi.org/10.1016/j.talanta.2016.06.004
[13]
Stubelius, A., Lee, S. and Almutairi, A. (2019) The Chemistry of Boronic Acids in Nanomaterials for Drug Delivery. AccountsofChemicalResearch, 52, 3108-3119. https://doi.org/10.1021/acs.accounts.9b00292
[14]
Aung, Y., Kristanti, A.N., Lee, H.V. and Fahmi, M.Z. (2021) Boronic-Acid-Modified Nanomaterials for Biomedical Applications. ACSOmega, 6, 17750-17765. https://doi.org/10.1021/acsomega.1c01352
[15]
Marinaro, W.A., Schieber, L.J., Munson, E.J., Day, V.W. and Stella, V.J. (2012) Properties of a Model Aryl Boronic Acid and Its Boroxine. JournalofPharmaceuticalSciences, 101, 3190-3198. https://doi.org/10.1002/jps.23207
[16]
Islam, T.M.B., Yoshino, K., Nomura, H., Mizuno, T. and Sasane, A. (2002) 1H and 11B NMR Study of P-Toluene Boronic Acid and Anhydride. AnalyticalSciences, 18, 363-366. https://doi.org/10.2116/analsci.18.363
[17]
Kinney, C.R. and Pontz, D.F. (1936) The Structure of the Organoboron Oxides. JournaloftheAmericanChemicalSociety, 58, 197-197. https://doi.org/10.1021/ja01293a005
[18]
Snyder, H.R., Kuck, J.A. and Johnson, J.R. (1938) Organoboron Compounds, and the Study of Reaction Mechanisms. Primary Aliphatic Boronic Acids. JournaloftheAmericanChemicalSociety, 60, 105-111. https://doi.org/10.1021/ja01268a033
[19]
Tokunaga, Y., Ueno, H., Shimomura, Y. and Seo, T. (2002) Formation of Boroxine: Its Stability and Thermodynamic Parameters in Solution. Heterocycles, 57, 787-790. https://doi.org/10.3987/com-02-9464
[20]
Tokunaga, Y. (2013) Boroxine Chemistry: From Fundamental Studies to Applications in Supramolecular and Synthetic Organic Chemistry. Heterocycles, 87, 991-1021. https://doi.org/10.3987/rev-13-767
Faniran, J.A. and Shurvell, H.F. (1968) Infrared Spectra of Phenylboronic Acid (Normal and Deuterated) and Diphenyl Phenylboronate. CanadianJournalofChemistry, 46, 2089-2095. https://doi.org/10.1139/v68-341
[23]
Soriano-Ursúa, M.A., Farfán-García, E.D., López-Cabrera, Y., Querejeta, E. and Trujillo-Ferrara, J.G. (2014) Boron-Containing Acids: Preliminary Evaluation of Acute Toxicity and Access to the Brain Determined by Raman Scattering Spectroscopy. NeuroToxicology, 40, 8-15. https://doi.org/10.1016/j.neuro.2013.10.005
[24]
Weiss, J.W.E. and Bryce, D.L. (2010) A Solid-State 11B NMR and Computational Study of Boron Electric Field Gradient and Chemical Shift Tensors in Boronic Acids and Boronic Esters. TheJournalofPhysicalChemistryA, 114, 5119-5131. https://doi.org/10.1021/jp101416k
[25]
Rose, M.E., Longstaff, C. and Dean, P.D.G. (1982) Gas Chromatography of Aromatic Boronic Acids: On-Column Derivatization. JournalofChromatographyA, 249, 174-179. https://doi.org/10.1016/s0021-9673(00)80245-9
[26]
Ji, Z., Petrovic, J. and Kott, L. (2021) Analysis of Boronic Compounds as Potential Mutagenic Impurities in Drug Substances by GC-MS. LCGC North America, 39, 588-592. https://www.chromatographyonline.com/view/analysis-of-boronic-compounds-as-potential-mutagenic-impurities-in-drug-substances-by-gc-ms
[27]
Lawrence, K., Flower, S.E., Kociok-Kohn, G., Frost, C.G. and James, T.D. (2012) A Simple and Effective Colorimetric Technique for the Detection of Boronic Acids and Their Derivatives. AnalyticalMethods, 4, 2215-2217. https://doi.org/10.1039/c2ay25346a
[28]
Salisbury, J.J., Georgian, W., Herr, M. and Buetti-Weekly, M. (2024) Validation of a Purity Method for a Suzuki-Miyaura Boronic Ester by Liquid Chromatography with Derivatization. JournalofChromatographyOpen, 5, Article ID: 100120. https://doi.org/10.1016/j.jcoa.2024.100120
[29]
Duval, F., Wardani, P.A., Zuilhof, H. and van Beek, T.A. (2015) Selective On-Line Detection of Boronic Acids and Derivatives in High-Performance Liquid Chromatography Eluates by Post-Column Reaction with Alizarin. JournalofChromatographyA, 1417, 57-63. https://doi.org/10.1016/j.chroma.2015.09.031
[30]
Pandiyan, P.J., Appadurai, R. and Ramesh, S. (2013) A High Throughput Analysis of Boronic Acids Using Ultra High Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometry. AnalyticalMethods, 5, 3386-3394. https://doi.org/10.1039/c3ay40542g
[31]
Dai, L., Gonzalez, J. and Zhang, K. (2022) A Simple Generic Method for Analyzing Water Sensitive Pinacol Boronate Compounds by Hydrophilic Interaction Liquid Chromatography. JournalofChromatographyOpen, 2, Article ID: 100036. https://doi.org/10.1016/j.jcoa.2022.100036
[32]
Haas, M.J., Blom, K.F. and Schwarz, C.H. (1999) Positive-Ion Analysis of Boropeptides by Chemical Ionization and Liquid Secondary Ionization Mass Spectrometry. AnalyticalChemistry, 71, 1574-1578. https://doi.org/10.1021/ac9810112
[33]
Flender, C., Leonhard, P., Wolf, C., Fritzsche, M. and Karas, M. (2010) Analysis of Boronic Acids by Nano Liquid Chromatography-Direct Electron Ionization Mass Spectrometry. AnalyticalChemistry, 82, 4194-4200. https://doi.org/10.1021/ac1004585
[34]
Wang, L., Dai, C., Burroughs, S.K., Wang, S.L. and Wang, B. (2013) Arylboronic Acid Chemistry under Electrospray Conditions. Chemistry—AEuropeanJournal, 19, 7587-7594. https://doi.org/10.1002/chem.201204290
[35]
Chidella, K.S., Dasari, V.B. and Jayashree, A. (2021) A High Sensitive LC-MS/MS Method for the Simultaneous Determination of Potential Genotoxic Impurities Carboxy Phenyl Boronic Acid and Methyl Phenyl Boronic Acid in Lumacaftor. AmericanJournalofAnalyticalChemistry, 12, 74-86. https://doi.org/10.4236/ajac.2021.123006
[36]
Mullangi, S., Ravindhranath, K. and Panchakarla, R.K. (2021) LC-MS/MS Method for the Quantification of Potential Genotoxic Impurity 4-Phenoxyphenyl-Boronic Acid in Ibrutinib. JournaloftheIranianChemicalSociety, 18, 1381-1389. https://doi.org/10.1007/s13738-020-02118-3
[37]
Berthelette, K.D., DeLoffi, M., Kalwood, J. and Haynes, K. (2024) Developing a Separation for Eleven Boronic Acids Using MaxpeakTM Premier Column Technology on an ArcTM HPLC System. WatersTM Corporation Application Note 720008307-en.
[38]
Maziarz, M. and Rainville, P.D. (2022) High Sensitivity Analysis of Potential Mutagenic Boronic Acids Using XevoTM TQ Absolute Tandem Quadrupole Mass Spectrometer with an ACQUITYTM Premier System. Waters？ Corporation Application Note 720007562-en.
