Characterization of Degradation Products of Guanfacine Hydrochloride API: Development and Validation of a Stability-indicating Reversed Phase UHPLC Method
Abstract
Guanfacine Hydrochloride API was subjected to forced degradation studies under various conditions of hydrolysis (acidic, alkaline, and neutral/water), oxidation, photolysis, and thermal. A short and simple, reverse phase UHPLC method was developed on a Shimadzu, Shimpack GIST, C18, (100 × 2.1) mm, 3.0 μm column. The gradient method consisted of 0.1% orthophosphoric acid as mobile phase A and acetonitrile as mobile phase B. The flow rate of the mobile phase was 0.3 mL/min. The method was validated using ICH guidelines considering the parameters solution stability, specificity, DL/QL, linearity, accuracy, precision and robustness. The drug was found highly sensitive to alkaline conditions and showed significant degradation. The drug was found sensitive to acidic degradation conditions. Slight degradation was observed in oxidative and water conditions. The drug was found to be stable in thermal and photolytic conditions. The mass compatible UHPLC method was prepared by simply substituting the orthophosphoric acid with formic acid in the mobile phase. Characterization of two major degradation products (DPs) was done. DP1 was characterized with LC–Q-TOF-MS/MS in combination with accurate mass measurements. DP2 was isolated and characterized with NMR, IR and HRMS spectroscopic techniques. The mechanisms of the formation of DPs were proposed.
Keywords:
Guanfacine, forced degradation, characterization, LC–Q-TOF-MS/MS, accurate mass measurements, stability indicatingDOI
https://doi.org/10.25004/IJPSDR.2023.150111References
Li Y, Henion J, Abbott R, Wang P. Dried blood spots as a sampling technique for the quantitative determination of guanfacine in clinical studies. Bioanalysis. 2011; 3(22):2501–2514.
Goparaju SM, Nandula YSM, Kothapalli CB, Challa BR, Awen BZ. Method development and validation of Guanfacine in rat plasma by liquid chromatography–tandem mass spectrometry: application to a pharmacokinetic study. J. Pharm. Anal. 2013; 3(6):472–480.
Inoue Y, Morita H, Nozawa K, Kanazu T. Metabolite profiling of guanfacine in plasma and urine of healthy Japanese subjects after oral administration of guanfacine extended‐release tablets. Biopharm. Drug. Dispos. 2019; 40:282–293.
Carrie Haglock, Carl Wolf, Alphonse Poklis. A novel method for the determination of guanfacine in urine by gas chromatography–mass spectrometry. J. Anal. Toxicol. 2008; 32(8):544-546.
Swearingen D, Pennick M, Shojaei A, Lyne A, Fiske K. A phase I, randomized, open-label, crossover study of the single-dose pharmacokinetic properties of guanfacine extended-release 1, 2 and 4 mg tablets in healthy adults. Clin. Ther. 2007; 29(4):617–625.
Guanfacine Hydrochloride monograph. USP 43-NF38, United States Pharmacopoeial Convention, Rockville, MD.
Sundari J, Amuthalakshmi S, Nalini CN. Review on analytical methods for quantification of ADHD drugs in human biological samples. Rev. Anal. Chem. 2020; 39:130–156.
Mishina EV, Marroum P, Bhattaram A, Gobburu J. Clinical Pharmacology Review. NDA 22-037, Guanfacine. US FDA. 2007.
Australian Public Assessment Report for Guanfacine (as hydrochloride). Intuniv. Shire Australia Pty Ltd. TGA. 2018.
International Conference on Harmonization (ICH) Guideline ICH Q1A(R2). Stability testing of new drug substances and products. IFPMA, Geneva, Switzerland, 2003.
Bakshi M, Singh S. Development of validated stability-indicating assay methods—critical review. J. Pharm. Biomed. Anal. 2002; 28:1011–1040.
International Conference on Harmonization (ICH) Guideline ICH Q1B. Stability Testing: Photostability Testing of New Drug Substances and Products. IFPMA, Geneva, Switzerland 1996.
International Conference on Harmonization (ICH) Guidelines ICH Q2. Validation of analytical procedures (R1). Toronto, 2005.
International Conference on Harmonization (ICH) Guidelines ICH Q14 (Draft version). Analytical procedure development. IFMPA, Geneva, 2022.
International Conference on Harmonization (ICH) Guidelines ICH Q3A (R2). Impurities in new drug substances. IFMPA, Geneva, 2006.
Desai R, Koradia S. Isolation, Identification and Characterization of Degradation Impurity of Atorvastatin in Fixed Dose Combination of Atorvastatin and Ezetimibe. Int. J. Pharm. Sci. Drug Res. 2019; 11(5): 187-193.
Singamsetti JCMKNNM, Korupolu RB, Gandham H, Geereddi MKR, Kaliyaperumal M, Rumalla CS, Mutha VSRNAK, Ivaturi R. Forced Degradation Studies of Nilotinib Hydrochloride: Isolation, Identification, and Characterization of Impurities. Int. J. Pharm. Sci. Drug Res. 2020;12(5):537-543.
Yerra NV, Dadinaboyina SB, Abbaraju LSSNV, Talluri MVNK, Thota JR. Identification and characterization of degradation products of indacaterol using LC/MS. Eur. J. Mass Spectrom. 2020; 26(6): 425-431.
Reddy GMK, Singamsetti JCMKNNM, Kaliyaperumal M, Doddipalla R, Ivaturi R, Rumalla CS, Korupolu RB, Babu BK. Degradation studies of levosimendan isolation, identification, and structure confirmation of stress degradation products using LCMS, mass mediated Prep-HPLC, NMR, HRMS, SFC and FTIR. J. Liq. Chromatogr. Relat. Technol. 2019; 3(1-2):1-12.
Saida SJ, Manikandan A, Kaliyaperumal M, Rumalla CS, Khan AA, Jayaraman VB, Yanaka R, Rao SV. Identification, isolation and characterization of dolutegravir forced degradation products and their cytotoxicity potential. J. Pharm. Biomed. Anal. 2019; 174: 588–594.
Prashanth S, Kumar AA, Madhu B, Sagar JV. LC and LC-MS study on stress decomposition behavior of paclitaxel and establishment of validated stability-indicating assay method. J. Pharm. Sci. Drug Res. 2011; 3(3):188-196.
Guerret M, Lavene D, Longchampt J, Kiger JL. Determination of guanfacine in biological fluids by electron-capture GLC. J. Pharm. Sci. 1979; 68(2):219-221.
Wahbi AAM, Bedair MM, Galal SM, Gazy AA. Spectrophotometric analysis of some guanidino drugs by acid-dye and charge-transfer complexation methods. J. Pharm. Biomed. Anal. 1993; 11(8):639-645.
Wahbi AAM, Bedair MM, Galal SM, Abdel-Hay MH, Gazy AA. Spectrofluorimetric determination of guanethidine sulphate, guanfacine hydrochloride, guanoclor sulphate and guanoxan sulphate in tablets and biological fluids, using benzoin. Microchim. Acta. 1993; 111:83-91.
Wolf CE, Kester-Florin SJ, Poklis A. A HPLC-MS method to detect and quantify guanfacine in urine. Clin. Chem. Lab. Med. 2012; 50(3):535–537.
Li Y, Henion J, Abbott R, Wang P. Semi-automated direct elution of dried blood spots for the quantitative determination of guanfacine in human blood. Bioanalysis. 2012; 4(12):1445–1456.
Li Y, Henion J, Abbott R, Wang P. The use of a membrane filtration device to form dried plasma spots for the quantitative determination of guanfacine in whole blood. Rapid Commun. Mass Spectrom. 2012; 26:1208–1212.
Li X, Li N, Sun X, Yang W, Dai Y, Xu J, Zhang W, Wang C, Wang S, Chen X. Development and validation of a simple, sensitive and accurate LC-MS/MS method for the determination of guanfacine, a selective α2A-adrenergicreceptor agonist, in plasma and its application to a pharmacokinetic study. Biomed. Chromatogr. 2013; 27: 1708–1713.
Goparaju SM, Nandula YSM, Kothapalli CB, Challa BR, Awen BZ. Method development and validation of guanfacine in rat plasma by liquid chromatography–tandem mass spectrometry: Application to a pharmacokinetic study. J. Pharm. Anal. 2013; 3(6):472–480.
Jebaliya H, Patel M, Dabhi B, Jadeja Y, Shah A. An isocratic assay method validation for the determination of five most potent antihypertensive drugs by a new generation liquid chromatographic technique. Chemistry & Biology Interface. 2016; 6(1):38-46.
Ahirrao VK, Sangale DR, Sonekara VS, Thorat VV, Marathe RP, Nawale RB, Pawar RP. Stability-indicating RP-HPLC method for determination of guanfacine hydrochloride in bulk drugs and in pharmaceutical dosage form. Int. J. Ind. Chem. 2011; 2(2):69-77.
Guanfacine Hydrochloride draft monograph. USP PF 48 (1), Jan. 2022. United States Pharmacopeial Convention, Rockville, MD.
FDA Foods and Veterinary Medicine Science and Research Steering Committee. Acceptance criteria for confirmation of identity of chemical residues using exact mass data within the office of foods and veterinary medicine. Memorandum. Department of Health & Human Services, Washington, D.C., United States, 2015.
Published
Abstract Display: 404 
PDF Downloads: 441 
