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.

Water For Injections (WFI) which is the main vehicle of Large Volume Parenterals (LVPs) should be free from trace amount of genotoxic impurities. This review gives emphasis on quantification of genotoxic trace metals during qualification of WFI in LVPs manufacturing unit. According to ICH guidelines, impurities related to drug substances are classified into three main categories: Organic impurities, inorganic (elemental) impurities and residual solvents. Within these categories, genotoxic impurities form a special case that poses a significant safety risk, even at low concentrations, because they may be mutagenic and are therefore potentially damaging to DNA. As a result they can lead to mutations or cause cancer. Chemical carcinogens most often directly or after xenobiotic metabolism, act as genotoxic causes to induce DNA damage. The roles of trace metals (some of which are either genotoxic or non-genotoxic) in cancer development and inhibition have a complex character and have raised many questions because of their essential and toxic effects on people's health. Trace metals such as cadmium, nickel, arsenic, beryllium and chromium (VI) have been recognized as human or animal carcinogens by International Agency for Research on Cancer (IARC). There are several genotoxic chemicals like residual solvent, impurities and trace metals present in Pharmaceuticals to form carcinogens. Regulatory body like FDA and EMEA has fixed up specific limits for these elemental impurities. The toxicity of an elemental impurity is related to its extent of exposure (bioavailability). In that sense, parenteral dosage forms has its most possibility to be bioavailable than the other dosage forms, the limit of genotoxic impurities are 10 times lower than that of oral dosage forms by United States Pharmacopoeia (USP). The present article is for the importance of identification and quantification of the trace amount of the metal genotoxic impurities in Water For Injection (WFI) during qualification (IQ, OQ, PQ) as a preventive measure to control the production and distribution of WFI for Large Volume Parenteral (LVP) production.