DEVELOPMENT OF TECHNOLOGY FOR OBTAINING MULTIPLE EMULSION OF OVARIAN LIQUID (CAVIAR SOL), ENSURING THE DURATION OF ITS STORAGE

Дефицит пищевого белка обусловливает актуальность развития отечественного производства пищевых белковых продуктов из биоресурсов вторичного происхождения. Овариальная жидкость (икорный золь) – вторичный продукт икорного производства – может быть использована как физиологически функциональный ингредиент для пищевых продуктов сбалансированного питания. Применяемые способы сохранности овариальной жидкости (ОЖ) приводят к изменению нативных свойств ее компонентов за счет температурного или химического воздействия. Высказано предположение, что капсулирование во множественную эмульсию ОЖ позволит сохранить ее нативные свойства и увеличить срок хранения. Цель настоящего исследования – разработка технологии получения множественной эмульсии ОЖ и установление эффективности новой формы продукта при хранении. Объектом исследования была ОЖ, полученная при заборе икры осетровых рыб с применением метода прижизненного получения. Множественную эмульсию готовили двухступенчатым способом. На первом этапе получали стандартную эмульсию вода–масло, которую затем использовали в качестве одной из фаз для множественной эмульсии вода–масло–ОЖ. Полученный продукт и контрольный образец (ОЖ, прошедшая только фильтрацию) размещали в чашках Петри и хранили в термостатируемой холодильной камере при температуре не ниже 4°С в течение 40 сут. Через каждые 5 сут в отобранных пробах определяли по методу Кьельдаля содержание белковых веществ, небелкового азота и азота летучих оснований. Установлено, что при хранении в ОЖ, прошедшей только фильтрацию, белок разрушался быстрее и его содержание через 40 сут снизилось на 34%, тогда как в ОЖ, заключенной во множественную эмульсию, этот показатель снизился только на 5,9%, что коррелирует со скоростью накопления продуктов распада белков в ОЖ после 40 сут хранения. Предложенный способ получения множественной эмульсии ОЖ позволяет обеспечить сохранность ее белкового комплекса в процессе хранения и разработать новый ассортимент физиологически функциональных продуктов с использованием отечественного биологически ценного пищевого ингредиента. The shortage of dietary protein determines the relevance of the development of domestic production of food protein products from bioresources of secondary origin. Ovarian liquid (caviar ash), or caviar sol, – a secondary product of caviar production – can be used as a physiologically functional ingredient for balanced nutrition foods. The applied methods of preservation of ovarian fluid (OF) lead to a change in the native properties of its components due to temperature or chemical exposure. It is suggested that encapsulation in a multiple emulsion of OF will preserve its native properties and increase the shelf life. The purpose of this study is to develop a technology for obtaining multiple OF emulsion and to establish the effectiveness of a new product form during storage. The object of the study was the OF obtained by sampling the caviar of sturgeon fish using the method of lifetime production. The multiple emulsion was prepared in a two-stage way. At the first stage a standard water–oil emulsion was obtained, which was then used as one of the phases for a multiple water–oil–OF emulsion. The resulting product and the control sample (OF that passed only filtration) were placed in Petri dishes and stored in a thermostatically controlled refrigerator at a temperature of at least 4°C for 40 days. Every 5 days the content of protein substances, non-protein nitrogen and nitrogen of volatile bases was determined in the selected samples by the Kjeldahl method. It was found that when stored in the OF that passed only filtration, the protein was destroyed faster and its content decreased by 34% after 40 days, whereas in the OF enclosed in a multiple emulsion, this indicator decreased by only 5.9%, which correlates with the rate of accumulation of protein breakdown products in the OF after 40 days of storage. The proposed method for obtaining a multiple emulsion of COOLANT makes it possible to ensure the safety of its protein complex during storage and to develop a new range of physiologically functional products using a domestic biologically valuable food ingredient.

Mass transfer of anti-cancer drug delivery to brain tumours by a multiple emulsion-based implant

The advanced use of a pH-responsive biomaterial-based injectable liquid implant for effective chemotherapeutic delivery in glioblastoma multiforme brain (GBM) tumour treatment is presented. As an implant, we proposed a water-in-oil-in-water multiple emulsion with encapsulated doxorubicin. The effectiveness of the proposed therapy was evaluated by comparing the cancer cell viability achieved in classical therapy (chemotherapeutic solution). The experimental study included doxorubicin release rates and consumption for two emulsions differing in drop sizes and structures in the presence of GBM-cells (LN229, U87 MG), and a cell viability. The results showed that the multiple emulsion implant was significantly more effective than classical therapy when considering the reduction in cancer cell viability: 85% for the emulsion-implant, and only 43% for the classical therapy. A diffusion-reaction model was adapted to predict doxorubicin release kinetics and elimination by glioblastoma cells. CFD simulations confirmed that the drug release kinetics depends on multiple emulsion structures and drop sizes.

DEVELOPMENT AND STATISTICAL OPTIMIZATION OF GASTRORETANTIVE FLOATING MICROSPHERES OF PREGABALIN PREPARED BY W/O/O MULTIPLE EMULSION METHOD

Objective: The aim of this study is to develop a gastroretentive microsphere of pregabalin using design of experiment (DoE) to decrease dosing frequency and increasing bioavailability. Methods: Pregabalin microsphere was prepared by W/O/O multiple emulsion method using a mixture of ethyl cellulose (EC) and polyvinyl pyrrolidone (PVP) as rate-controlling polymer. Mixed solvent system comprising of dichloromethane (DCM) and acetonitrile (ACN) and light liquid paraffin was chosen as primary and secondary oil phase respectively. Taguchi design was employed for factor screening and Box Behnken design was used for the optimisation of critical process parameters. Results: Taguchi design revealed that polymer: drug, DCM: ACN and PVP: EC is the critical factor for the preparation of microspheres. The optimized formulation was prepared using polymer: drug (4.95:1), DCM: ACN (53.76: 46.24) and PVP: EC (2:5) which showed mean particle size of 203.34±4.82 µm, practical yield of 87.52±2.91 %, encapsulation efficiency of 96.43±3.14 %, floating ability up to 90.42±1.64 % and T60% of 332.81±5.84. Drug release from microsphere followed Higuchi Kinetic model. Conclusion: In a nutshell, microspheres with excellent flowability and great encapsulation efficiency were successfully developed. These can be useful in improving patient compliance by reducing frequent dosing.

Preparation of Neuropeptide Nanoparticle and Its Mechanism in Corneal Nerve Regeneration in Substance P-Neurokinin 1 Receptor Signaling Pathway

The multiple emulsion method was applied to prepare the 1.5 mL solution of ciliary neurotrophic factor (CNTF), and dichloromethane solution with 2.5% polylactic acid (PLA) was added into the CNTF solution during the colostrum process, and then, polyethylene glycol solution was added into the mixed solution under ultrasonic conditions to form multiple emulsion; finally, it was stirred magnetically and centrifuged to obtain CNTF nanoparticles. The transmission electron microscope (TEM) was applied to characterize the prepared nanoparticles that detected their drug loading rate, encapsulation rate, and their drug release in vitro and in vivo. In the demonstration of neuropeptide nanoparticles participating in corneal nerve regeneration in the substance p-neurokinin 1 receptor (SP-NK-1R) signaling pathway, it was to detect the gene expression in the trigeminal ganglion cells based on the different groups like normal group, advanced glycation end products (AGE)/Stattic group, and AGE + CNTF/Stattic + CNTF group. In the characterization experiment of CNTF nanoparticles, their surfaces were smooth, they evenly distributed, and the average drug loading rate reached 1.27% ± 0.032%; the nanoparticles were able to continuously release CNTF molecules within 30 hours; what’s more, CNTF group (low concentration to high concentration) and CNTF nanoparticles group (low concentration to high concentration) could both enhance the expression of mouse optic nerve cells. Compared to AGE group, the expressions of substance p-signal transducer and activator of transcription 3 (p-STAT3) in trigeminal ganglion cells increased after adding CNTF and CNTF nanoparticles, respectively (P < 0.05); and the expression of p-STAT3 increased remarkably after the addition of CNTF nanoparticles (P < 0.05), with an obvious increase on the growth of nerve axon. On the other hand, it had the same results as the above, compared with Stattic group.