Seabuckthorn (Hippophae rhamnoides L.) is considered a multipurpose crop. Different parts of the plant are marked with a unique composition of nutrients and biologically active substances (soluble sugars, fatty acids, vitamins, organic acids, etc.). The study of biochemical composition is necessary for successful breeding and widening seabuckthorn utilization in medicine, food, and cosmetics industries. The paper presents the integrated biochemical features of seabuckthorn berries bred by the Lisavenko Institute in Barnaul, Russia (3 varieties and 58 hybrids). Biochemical analysis was carried out according to common research methods. We established the variability for such substances as soluble solids, sugars, vitamin C, titratable acids, oils, and sugar acids. Significant variability for sugars content (22%), vitamin C content (32%), and sugar-acid index (36%) was revealed. According to the assessment results, we selected 12 promising genotypes. We also identified genotypes combining maximum levels of 1–3 biochemical features.