Food Processing: Techniques and Technology Food Processing: Techniques and Technology Техника и технология пищевых производств 2074-9414 2313-1748 72923 10.21603/2074-9414-2023-4-2464 ОРИГИНАЛЬНАЯ СТАТЬЯ ORIGINAL ARTICLE ОРИГИНАЛЬНАЯ СТАТЬЯ Comprehensive Methodological Approach to Determining Lipids in Clams Комплексный методический подход в определении липидов моллюсков https://orcid.org/0000-0001-8468-8372 Бородина Александра Валентиновна Borodina Alexandra V. borodinaav@mail.ru https://orcid.org/0000-0003-0372-2458 Веляев Юрий Олегович Velyaev Yuri O. https://orcid.org/0009-0003-5962-1899 Осокин Александр Рузвельтович Osokin Alexander R. Институт биологии южных морей имени А.О. Ковалевского РАН Севастополь Россия Kovalevsky Institute of Biology of Southern Seas, Russian Academy of Sciences Sevastopol Russian Federation Севастопольский государственный университет Севастополь Россия Sevastopol State University Sevastopol Russian Federation Севастопольский государственный университет Севастополь Россия Sevastopol State University Sevastopol Russian Federation 30 12 2023 30 12 2023 53 4 662 671 17 03 2023 06 06 2023 https://fptt.ru/en/issues/22269/22205/

Морские двустворчатые моллюски характеризуются повышенным содержанием полиненасыщенных жирных кислот. В Средиземном и Чёрном морях к потенциально промысловым относится моллюск Cerastoderma glaucum, который содержит омега-3, омега-6 и омега-9 жирные кислоты. Однако липидный состав этого вида гидробионтов слабо изучен. При определении общих липидов, их классов и состава жирных кислот стандартные методы часто необходимо адаптировать к особенностям объекта исследования и имеющейся приборной базе. Цель работы – предложить вариант комплексной методики определения липидов гидробионтов на примере двустворчатого моллюска C. glaucum. Объектом исследования была сумма мягких тканей моллюска C. glaucum, распространенного в зоне псевдолиторали на песочно-иловом грунте Севастопольского побережья Чёрного моря. Применяли хроматографические методы установления общих липидов, разделения их на классы (фосфолипиды, моноглицериды, диглицериды, стеролы, триацилглицерины) и последующего денситемтрического определения с использованием планшетного сканера и программы ТСХ менеджер 4.0.2.3D. Исследование состава жирных кислот с помощью хромато-масс-спектрометрического метода проводили для общих липидов. В работе привели методические указания по адаптации известных методов в липидологии по определению общих липидов, их классов и жирнокислотного состава в тканях моллюсков C. glaucum. Описали метод определения общих липидов. Показали подготовку оборудования и реактивов для разделения общих липидов на классы методом многомерной тонкослойной хроматографии. Представили авторскую схему хроматографических ванн для осуществления ступенчатого разделения. Провели денситометрическое измерение. Привели примеры хроматограммы жирных кислот и масс-спектров. Предложили вариант проведения пробоподготовки к определению жирных кислот в общих липидах методом газовой хроматографии, который отличается минимальной потерей нативной структуры веществ и является более мягким по сравнению с широко применяемым методом дериватизации проб. Применение предлагаемого метода для определения липидов моллюсков является экономичным и менее затратным по времени и реактивам. Метод рекомендуется использовать для небольших лабораторий, занимающихся фундаментальными исследованиями энергетики организмов или в прикладных целях для сравнительных анализов гидробионтов.

Marine bivalves are rich in polyunsaturated fatty acids. Cerastoderma glaucum is a potentially commercial sea cockle that inhabits the Mediterranean and the Black Seas. This bivalve mollusk contains omega-3, omega-6, and omega-9 fatty acids. However, its lipid composition remains understudied. When determining total lipids, their classes, and fatty acid composition, standard methods often have to be adapted to the object in hand and tools available. The research objective was to develop a complex lipid analysis method for aquatic organisms. The study featured total soft tissues of C. glaucum harvested from the pseudolittoral zone on the sand and silt soil of the Sevastopol coast of the Black Sea. The chromatographic methods made it possible to identify total lipids and classify them into phospholipids, monoglycerides, diglycerides, sterols, and triacylglycerols. The subsequent densimetric determination involved a flatbed scanner and the TLC Manager 4.0.2.3D software. The fatty acid composition for total lipids was studied using the chromatography-mass spectrometric method. The existing methods in lipidology were adapted for determining total lipids, their classes, and the fatty acid composition of total lipids in C. glaucum. The article introduces a detailed description of the method for determining total lipids, as well as of how to prepare equipment and reagents to classify common lipids using multidimensional thin layer chromatography. It also contains an authentic scheme of chromatographic baths for stepwise separation, densitometric measurements, and examples of fatty acid chromatograms and mass spectra. The new sample preparation method for determining fatty acids in total lipids by gas chromatography demonstrated a minimal loss in native structure and proved to be less aggressive than standard methods of sample derivatization. The new method for lipid analysis of clam tissues appeared to be economical, less time-consuming, and reagent-intensive. It can be recommended for small laboratories engaged in bioenergetics or comparative analyzes of aquatic organisms.

 Общие липиды классы липидов тонкослойная хроматография денситограмма хроматограмма хромато-масс-спектрометрия моллюски Total lipids lipid classes thin layer chromatography densitogram chromatogram fatty acids gas chromatography/mass spectrometry shellfish Работа выполнена по теме № 121041400077-1 государственного задания Института биологии южных морей имени А. О. Ковалевского РАН (ФИЦ ИнБЮМ) «Функциональные, метаболические и токсикологические аспекты существования гидробионтов и их популяций в биотопах с различным физико-химическим режимом». В работе использовано оборудование ЦКП «Молекулярная структура вещества» Севастопольского государственного университета (СевГУ). The work part of State Assignment to A.O. Kovalevsky Institute of Biology of the Southern Seas of RAS (IBSS) , research topic No. 121041400077-1: Functional, metabolic, and toxicological aspects of hydrobionts and their populations in biotopes with different physical and chemical modes. The research was conducted on the premises of Core Facilities Centre of Molecular Matter Structure, Sevastopol State University.

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