Key Biochemical Parameters and Enzyme Activity of Umbilical Cord Blood (Chernivtsi, Ukraine, 2024–2025)

Umbilical cord blood (158 samples) was collected from May 2024 to January 2025 at the Chernivtsi Regional Perinatal Center (Chernivtsi, Ukraine) from mothers at risk of developing hemolytic disease of the newborn (HDN) without signs of the disease, namely mothers with an Rh(D)-negative blood factor regardless of blood group, as well as mothers with blood group O (I) and an Rh(D)-positive blood factor.The study was conducted in accordance with the ethical principles for medical research involving human subjects set forth in the Declaration of Helsinki (1964–2013), ICH GCP (1996), EEC Directive No. 609 (dated November 24, 1986), and Order of the Ministry of Health of Ukraine No. 690 dated September 23, 2009, and was approved by the Biomedical Ethics Committee of Bukovinian State Medical University (Chernivtsi, Ukraine). Written informed consent was obtained from all mothers for the collection and use of umbilical cord blood samples for research purposes. In the collected umbilical cord blood samples, the AB0 blood group and Rh factor were determined, and the following biochemical parameters were measured: total bilirubin (µmol/L), determined using the Malloy–Evelyn method with a sulfodiazonium salt direct bilirubin (µmol/L), determined using the vanadate oxidation method, with the formation of biliverdin total protein (g/L), determined using the biuret method albumin (g/L), determined using bromocresol green (BCG) albumin-globulin ratio is calculated by the formula albumin/(total protein- albumin) glucose (mmol/L), determined using the hexokinase method urea (mmol/L), determined using a kinetic enzymatic method with urease creatinine (µmol/L), determined using the modified Jaffe method without deproteinization alanine aminotransferase (U/L) activity, assessed based on the amount of oxidized NAD⁺ formed in the corresponding chemical conversion reactions in the absence of pyridoxal phosphate aspartate aminotransferase (U/L) activity, assessed based on the amount of oxidized NAD⁺ formed in the corresponding chemical conversion reactions in the absence of pyridoxal phosphate γ-glutamyltransferase (U/L) activity, determined using a kinetic method with L-γ-glutamyl-3-carboxy-4-nitroanilide, yielding 5-amino-2-nitrobenzoate as the reaction product alkaline phosphatase (U/L) activity, determined kinetically based on the formation of 4-nitrophenol total lactate dehydrogenase (U/L) activity, determined in accordance with the recommendations of the German Society for Clinical Chemistry (DGKL), based on the amount of oxidized NAD⁺ uric acid (µmol/L), determined using an enzymatic colorimetric method with uricase and peroxidase