А. Основы спектрофотометрии

Многие молекулы поглощают свет в видимой или ультрафиолетовой области спектра. Это свойство можно использовать для определения концентраций. Величина поглощения зависит от типа и концентрации вещества, а также от длины волны используемого света. Поэтому применяют монохроматический свет, то есть свет определённой длины волны, который можно выделить из белого света с помощью монохроматора. Монохроматический свет интенсивности l0 проходит через прямоугольную ячейку из стекла или кварца (кювету), в которой находится раствор поглощающего вещества.

Интенсивность I выходящего света, ослабленного поглощением, измеряется с помощью детектора. Поглощение света (А) раствора (оптическая плотность) определяется как отрицательный логарифм отношения l/l0. Закон Ламберта-Бера гласит, что А пропорциональна концентрации (c) вещества и толщине (d) слоя раствора. Коэффициент экстинкции ε зависит, как было указано выше, от типа вещества и длины волны.


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