Simultaneous Determination of Phytonadione and its Degradants by HPLC in Injectable Emulsion Formulation Used to Treat VKDB in Newborns
Abstract
Phytonadione, also known as Phytomenadione or Vitamin K1 is used as an Injectable emulsion is used to Newborns for the precautionary treatment for Vitamin K Deficiency Bleeding (VKDB) during birth. In the current study, A Gradient, Sensitive & Cost effective HPLC method has been developed to determine and quantify the degradants of Phytonadione and Benzyl Alcohol into Phytonadione Injectable Emulsion. The chromatographic separation was performed by GL Sciences Inert sustain HP™ C18 (250 × 4.6 mm, 3.0 μ) column. The degradants were well separated by a gradient program started with the mixture of 25 mM Ammonium Acetate in water as buffer, pH 3.5 and Methanol in the ratio of 40: 60 V/V at the flow rate of 1.0 mL min‑1 and UV detection was performed at 254 nm. The degradation products from Phytonadione and Benzyl Alcohol were well resolved from the main peaks and its other impurities by the developed method. The method was validated by complying specificity/selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy and precision following ICH Q2 (R1). The developed method in this study could be applied for the analysis of routine quality control related substances of Phytonadione Injectable Emulsion, since there is no official monograph.
Keywords:
Phytomenadione, Vitamin K, Method validation, Impurities, Degradants
References
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22. Zhang Y, Bala V, Mao Z, Chhonker YS, Murry DJ. A concise review of quantification methods for determination of vitamin K in various biological matrices. Journal of pharmaceutical and biomedical analysis. 2019 May 30; 169:133-41.
23. Xue M, Xue Y, Yan C, Wang Y. NP-HPLC method for determining phytonadione and its impurities. Journal of Liquid Chromatography & Related Technologies. 2016 Nov 7;39(17-18):799-805.
2. Shah BA and Parikh K. HPLC Determination of Phytomenadione in Drop. Current Research in Pharmaceutical Sciences. 2013; 02:48-52.
3. Wakabayashi H, Onodera K, Yamato S, Shimada K. Simultaneous determination of vitamin K analogs in human serum by sensitive and selective high-performance liquid chromatography with electrochemical detection. Nutrition. 2003 Jul 1;19(7-8):661-5.
4. Shearer MJ. Vitamin K deficiency bleeding (VKDB) in early infancy. Blood reviews. 2009 Mar 1;23(2):49-59.
5. Label of Vitamin K1 Phytonadione Injection, Emulsion; [cited 2021 Jul 26]. Available from: https://fda.report/DailyMed/E8808230-2C44-44C6-8CAB-8F29B6B34051.pdf
6. Why do Newborns Need a Vitamin K Shot? By Michelle M. Karten, MD [cited 20108 April]. Available from: https://kidshealth.org/en/parents/vitamin-k-shot.html
7. Forchielli ML, Conti M, Motta R, Puggioli C, Bersani G. Phytonadione Content in Branded Intravenous Fat Emulsions: A Comparative Study of 6 Products with Different Fat Sources Using Liquid Chromatography–Mass Spectrometry. Journal of Parenteral and Enteral Nutrition. 2017 Mar;41(3):489-95.
8. Gong BY, Ho JW. Simultaneous separation and detection of ten common fat-soluble vitamins in milk. Journal of liquid chromatography & related technologies. 1997 Sep 1;20(15):2389-97.
9. Haroon Y, Bacon DS, Sadowski JA. Liquid-chromatographic determination of vitamin K1 in plasma, with fluorometric detection. Clinical chemistry. 1986 Oct 1;32(10):1925-9.
10. ICH, 2005. International Conference on Harmonisation (ICH) Q2 (R1): Validation of Analytical Procedures: Test and Methodology, Geneva, Switzerland.
11. "Institute of Medicine (US) Panel on Micronutrients. Dietary Reference Intakes for Vitamin A, Vitamin K, Arsenic, Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybdenum, Nickel, Silicon, Vanadium, and Zinc". NCBI. National Academies Press (US). Retrieved 25 November 2021.
12. Jeong IS, Gu SY, Park KH, Lee SY, Kim SG. A simultaneous determination and monitoring of vitamin K1 (phylloquinone) and vitamin K2 (menaquinone) in vegetable drinks and natto sold on the Korean market. Journal of Food Measurement and Characterization. 2022 Feb;16(1):248-57.
13. Kashani HN, Moghaddam FA, Mehramizi A. HPLC–UV CHROMATOGRAPHY DETERMINATION OF BENZALDEHYDE ARISING FROM BENZYL ALCOHOL USED AS PRESERVATIVE IN INJECTABLE FORMULATIONS. Asian Journal of Pharmaceutical and Clinical Research. 2012;5(2):98-100.
14. Langenberg JP, Tjaden UR. Improved method for the determination of vitamin K1 epoxide in human plasma with electrofluorimetric reaction detection. Journal of Chromatography A. 1984 Apr 27; 289:377-85.
15. Langenberg JP, Tjaden UR. Determination of (endogenous) vitamin K1 in human plasma by reversed-phase high-performance liquid chromatography using fluorometric detection after post-column electrochemical reduction: comparison with ultraviolet, single and dual electrochemical detection. Journal of Chromatography B: Biomedical Sciences and Applications. 1984 Jan 1; 305:61-72.
16. Lennon C, Davidson KW, Sadowski JA, Mason JB. The vitamin K content of intravenous lipid emulsions. Journal of Parenteral and Enteral Nutrition. 1993 Mar;17(2):142-4.
17. Nakata Y, Tsuchida E. [20] Determination of Vitamin-K1 in photodegradation products by gas—liquid chromatography. In Methods in Enzymology 1980 Jan 1 (Vol. 67, pp. 148-160). Academic Press.
18. Nannapaneni NK, Jalalpure SS, Muppavarapu R, Sirigiri SK. A sensitive and rapid UFLC-APCI-MS/MS bioanalytical method for quantification of endogenous and exogenous Vitamin K1 isomers in human plasma: Development, validation and first application to a pharmacokinetic study. Talanta. 2017 Mar 1; 164:233-43.
19. Patel SA, Parikh PK. Development and validation of spectrophotometric methods fordetermination of phytomenadione in injection. Current Research in Pharmaceutical Sciences. 2012 Apr 7:26-30.
20. Rébufa C, Artaud J. Analytical Determination of Phylloquinone (Vitamin K1) in Olive Oils. Comparison with Other Vegetable Oils. European journal of lipid science and technology. 2018 Jun;120(6):1700527.
21. Moussa F, Depasse F, Lompret V, Hautem JY, Girardet JP, Fontaine JL, Aymard P. Determination of phylloquinone in intravenous fat emulsions and soybean oil by high-performance liquid chromatography. Journal of Chromatography A. 1994 Apr 1;664(2):189-94.
22. Zhang Y, Bala V, Mao Z, Chhonker YS, Murry DJ. A concise review of quantification methods for determination of vitamin K in various biological matrices. Journal of pharmaceutical and biomedical analysis. 2019 May 30; 169:133-41.
23. Xue M, Xue Y, Yan C, Wang Y. NP-HPLC method for determining phytonadione and its impurities. Journal of Liquid Chromatography & Related Technologies. 2016 Nov 7;39(17-18):799-805.
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How to Cite
Mohabbat Ullah, Md. Sohel Rana, and Md. Shahnowaj Bhuiyan. “Simultaneous Determination of Phytonadione and Its Degradants by HPLC in Injectable Emulsion Formulation Used to Treat VKDB in Newborns”. Current Research in Pharmaceutical Sciences, Apr. 2023, pp. 44-51, doi:10.24092/CRPS.2023.130104.
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Research Articles
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