Inskeep W P, Bloom P R
Department of Soil Science, University of Minnesota, St. Paul, Minnesota 55108.
Plant Physiol. 1985 Feb;77(2):483-5. doi: 10.1104/pp.77.2.483.
We found inconsistencies in the commonly used data for chlorophyll analysis in 80% acetone. Recently developed extinction coefficients for chlorophyll b in N,N-dimethylformamide (DMF) based on values from 80% acetone are low as a result of these inconsistencies. We determined extinction coefficients of chlorophyll a (Chl a) and chlorophyll b (Chl b) in DMF for wavelengths of 618 to 665 nanometers. The simultaneous equations necessary for quantifying Chl a, Chl b, or total Chl in DMF in the absence of other chlorophyllous pigments are: Chl a = 12.70A(664.5) - 2.79A(647); Chl b = 20.70 A(647) - 4.62A(664.5); total Chl = 17.90A(647) + 8.08A(664.5), where A = absorbance in 1.00 centimeter cuvettes and Chl = milligrams per liter.N,N-Dimethylformamide is a very convenient solvent for Chl extraction since it is effective on intact plant parts and Chl is quite stable in DMF. There was no difference in the amount of Chl extracted when plant tissue was stored for 1 or 3 days at three temperatures, with or without solvent added.
我们发现,用于80%丙酮中叶绿素分析的常用数据存在不一致性。由于这些不一致性,基于80%丙酮的值最近开发的N,N-二甲基甲酰胺(DMF)中叶绿素b的消光系数较低。我们测定了DMF中叶绿素a(Chl a)和叶绿素b(Chl b)在618至665纳米波长下的消光系数。在不存在其他叶绿素色素的情况下,定量DMF中Chl a、Chl b或总Chl所需的联立方程为:Chl a = 12.70A(664.5) - 2.79A(647);Chl b = 20.70 A(647) - 4.62A(664.5);总Chl = 17.90A(647) + 8.08A(664.5),其中A = 1.00厘米比色皿中的吸光度,Chl = 毫克/升。N,N-二甲基甲酰胺是一种非常方便的叶绿素提取溶剂,因为它对完整的植物部分有效,并且叶绿素在DMF中相当稳定。在三种温度下,添加或不添加溶剂时,植物组织储存1天或3天,叶绿素提取量没有差异。