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在弱碱性介质中，乳化剂－ＯＰ存在下，羟基萘酚蓝（ＨＮＢ）和钙生成蓝色２∶１ 络合物，λｍａｘ＝ ６５０ｎｍ ，ε＝ １．８６×１０４Ｌ·ｍ ｏｌ－ １·ｃｍ － １，钙含量在０－ ４０μｇ／２５ｍ Ｌ内符合比尔定律，本方法用于烟叶中钙的测定，结果令人满意。
Hydroxynaphthol blue (HNB) and calcium generated a blue 2: 1 complex in a slightly alkaline medium with emulsifier-OP, λmax = 650 nm, ε = 1.86 × 104 L · mol-1 · cm - 1. Beer ’s law was obeyed in the range of 0-40μg / 25m L. The method was applied to the determination of calcium in tobacco leaves with satisfactory results.