Термодинамический и структурный анализ формирования и функциональности пищевых биополимерных наносистем для доставки липофильных биологически активных веществ тема диссертации и автореферата по ВАК РФ 02.00.04, кандидат химических наук Григорович, Надежда Викторовна

• Григорович, Надежда Викторовна
• кандидат химических наук
• 2012

Оглавление диссертации кандидат химических наук Григорович, Надежда Викторовна

1. СПИСОК СОКРАЩЕНИЙ И УСЛОВНЫХ ОБОЗНАЧЕНИЙ

2. ВВЕДЕНИЕ

3. ОБЗОР ЛИТЕРАТУРЫ

3.1. Биологически активные вещества, относящиеся к классу нутрицевтиков

3.2. Процессы инкапсулирования и системы доставки

3.3. Системы доставки на основе пищевых биополимеров

3.3.1. Биополимерные гели и гелевые частицы

3.3.2. Биополимеры как эмульгаторы при эмульсионном инкапсулировании

3.3.3. Смеси и комплексы белков и полисахаридов

3.4. Факторы, влияющие на биодоступность биологически активных веществ в пищеварительной системе человека 39 Заключение по литературному обзору

4. ЦЕЛИ И ЗАДАЧИ РАБОТЫ

5. ЭКСПЕРИМЕНТАЛЬНАЯ ЧАСТЬ

5.1. Объекты исследования

5.1.1. Казеинат натрия

5.1.2. Мальто декстрины

5.1.3. Фосфатидилхолин

5.2. Методы исследования

5.2.1. Приготовление растворов биополимеров

5.2.2. Приготовление ковалентных коньюгатов казеината натрия с мальтодекстринами (Б + МД)

5.2.3. Приготовление водных растворов фосфатидилхолина

5.2.4. Приготовление комплексов казеината натрия, мальтодекстринов, простых смесей (Б + МД) и ковалентных коньюгатов (Б + МД) с фосфатидилхолином

5.2.5. Рефрактометрия

5.2.6. Оценка степени связывания казеината натрия с мальтодекстринами в ковалентных коньюгатах

5.2.7. Количественная оценка ФТДХ, связавшегося с казеинатом натрия, ковалентными коньюгатами, мальтодекстринами и простыми смесями казеината натрия с мальтодекстринами

5.2.8. Оценка степени растворимости казеината натрия в водных растворах ковалентных коньюгатов белка с мальтодекстринами

SA2 и MD10 и их комплексов с ФТДХ

5.2.9. Вискозиметрия в разбавленных растворах

5.2.10. Статическое многоугловое лазерное светорассеяние

5.2.11. Динамическое лазерное светорассеяние

5.2.12. Оценка пенообразующей способности биополимеров, ковалентных коньюгатов и их комплексов с ФТДХ

5.2.13. Оценка степени окисления чистого ФТДХ и его комплексов с казеинатом натрия, мальтодекстринами, простыми смесями и ковалентными коньюгатами (Б + МД)

5.2.14. Оценка действия ферментов желудочно-кишечного тракта на комплексы ФТДХ с ковалентными коньюгатами (Б + МД) invitro

5.2.15. Дифференциальная сканирующая калориметрия 78 6. РЕЗУЛЬТАТЫ И ИХ ОБСУЖДЕНИЕ 79 6.1. Структурные и термодинамические особенности комплексов казеината натрия с полиненасыщенным ФТДХ, определяющие их многофункциональность, как белковых наноконтейнеров для доставки ФТДХ и как структурообразующих и стабилизирующих агентов для коллоидных систем

6.1.1. Структурные и термодинамические параметры супрамолекулярных комплексов (казеинат натрия + ФТДХ), определённые методом многоуглового лазерного светорассеяния

6.1.2. Защита полиненасыщенного ФТДХ от окисления кислородом воздуха в супрамолекулярных комплексах с казеинатом натрия

6.1.3. Высвобождение ФТДХ из супрамолекулярных комплексов с казеинатом натрия в модельных условиях их переваривания в желудочно-кишечном тракте т-уИго.

6.1.4. Пенообразующая способность супрамолекулярных комплексов ФТДХ с казеинатом натрия

6.1.5. Состояние липосом ФТДХ в комплексе с казеинатом натрия

6.2. Структурные и термодинамические особенности ковалентных коньюгатов казеината натрия с мальтодекстринами, лежащие в основе их традиционной и новой функциональности

6.2.1. Растворимость ковалентных коньюгатов казеината натрия с мальтодекстринами в водной среде.

6.2.2. Пенообразующие свойства ковалентных коньюгатов казеината натрия с мальтодекстринами

6.3. Структурные и термодинамические особенности комплексов ковалентных коньюгатов (казеинат натрия + мальтодекстрин) с ФТДХ, определяющие их новую функциональность, как наноконтейнеров для доставки ФТДХ

6.3.1. Состояние липосом ФТДХ в комплексе с ковалентными коньюгатами казеината натрия и мальтодекстринов

6.3.2. Растворимость комплексов ковалентных коньюгатов (казеинат натрия + мальтодекстрин) с ФТДХ в водной среде

6.3.3. Окисление ФТДХ в комплексе с казеинатом натрия, мальтодекстринами, ковалентными коньюгатами Б + БА2 и Б Мй10 , простыми смесями Б + БА2 иБ + М£>

6.3.4. Пенообразующие свойства комплексов ковалентных коньюгатов (казеинат натрия + мальтодекстрин) с ФТДХ

6.3.5. Высвобождение ФТДХ из комплексов с ковалентными коньюгатами (Б + МД) в модельных условиях их переваривания в желудочно-кишечном тракте т-укго

ВЫВОДЫ

Заключение диссертации по теме «Физическая химия», Григорович, Надежда Викторовна

выводы

1. Впервые установлены ключевые структурные факторы комплексов ФТДХ с казеинатом натрия и ковалентными коньюгатами казеината натрия с мальтодекстринами, контролирующие эффективную защиту полиненасыщенного ФТДХ от окисления. Ими являются: плотность комплексных частиц, которая должна быть выше 2 мг/мл для 100% защиты ФТДХ от окисления, и степень ассоциации биополимеров в комплексных частицах при их плотности ниже 2 мг/мл. Найденный же немного более низкий, по сравнению с белком, защитный эффект ковалентных коньюгатов казеината натрия с мальтодекстринами по отношению к окислению ФТДХ при нейтральном рН объясняется более низкой чем у белка плотностью образованных комплексных частиц в этом случае.

2. Впервые было показано, что бислой модельного фосфатидилхолина (ДФТДХ) сохранял свою целостность в комплексе с казеинатом натрия и даже становился более упорядоченным в комплексе с ковалентными коньюгатами с увеличением числа присоединённых к белку гидрофильных молекул мальтодекстринов.

3. Впервые установлено, что уровень растворимости ковалентных коньюгатов казеината натрия с мальтодекстринами тем выше в области ИЭТ белка, чем большее количество молекул мальтодекстрина ковалентно присоединяется к белку. При этом роль степени полимеризации (ДЭ) мальтодекстринов проявляется в пространственных ограничениях при ковалентном присоединении мальтодекстринов к белку, т.е. чем выше ДЭ и ниже степень полимеризации, тем больше молекул МД может быть присоединено к белку. Определено, что 100% растворимость белка в коньюгатах достигалась при пятикратном весовом избытке мальтодекстринов в системе. При этом наблюдаемое увеличение растворимости ковалентных коньюгатов по сравнению с белком объясняется значительным ростом их термодинамического сродства к водной среде.

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4. Впервые установлено, что как ковалентное присоединение большого числа молекул мальтодекстрина к частицам казеината натрия, так и дополнительное присоединение большого числа молекул ФТДХ к частицам ковалентных коньюгатов приводит к ярко выраженной диссоциации исходных частиц белка и к изменению гидрофобно-гидрофильного баланса свойств их поверхности.

5. Впервые установлено, что оптимальное термодинамическое сродство к водной среде комплексных частиц ФТДХ с казеинатом натрия и с ковалентными коньюгатами казеината натрия и мальтодекстринов определяет максимальную стабильность пен, сформированных этими частицами, во времени.

6. Впервые установлено, что практически полное высвобождение ФТДХ из его комплексных частиц с казеинатом натрия наблюдается на стадии их переваривания в желудке под действием пепсина in vitro. Также впервые было найдено, что чем большее число молекул мальтодекстринов ковалентно пришито к казеинам, тем важнее роль амилазы, действующей в тонком кишечнике для эффективного высвобождения ФТДХ из его комплексов с ковалентными коньюгатами этих биополимеров in vitro. При этом, варьируя биополимерный состав комплексов, мы можем добиться контролируемого высвобождения ФТДХ в определённом отделе ЖКТ человека.

7. Таким образом, впервые было показано, что наночастицы казеината натрия и ковалентных коньюгатов казеината натрия с мальтодекстринами являются эффективными системами доставки для ФТДХ, благодаря их следующим свойствам: а) высокой степени связывания ФТДХ (> 90%); б) контролируемого высвобождения ФТДХ под действием ферментов ЖКТ in vitro; в) высокой степени защиты ФТДХ от окисления кислородом воздуха в условиях авто- и ускоренного окисления.

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