Romero J M, Lara C, Guerrero M G
Instituto de Bioquímica Vegetal y Fotosíntesis, Universidad de Sevilla y C.S.I.C., Spain.
Biochem J. 1989 Apr 15;259(2):545-8. doi: 10.1042/bj2590545.
A sensitive procedure has been developed for the determination of intracellular nitrate. The method includes: (i) preparation of cell lysates in 2 M-H3PO4 after separation of cells from the outer medium by rapid centrifugation through a layer of silicone oil, and (ii) subsequent nitrate analysis by ion-exchange h.p.l.c. with, as mobile phase, a solution containing 50 mM-H3PO4 and 2% (v/v) tetrahydrofuran, adjusted to pH 1.9 with NaOH. The determination of nitrate is subjected to interference by chloride and sulphate when present in the samples at high concentrations. Nitrite also interferes, but it is easily eliminated by treatment of the samples with sulphamic acid. The method has been successfully applied to the study of nitrate transport in the unicellular cyanobacterium Anacystis nidulans.
已开发出一种用于测定细胞内硝酸盐的灵敏方法。该方法包括：(i) 通过硅油层快速离心将细胞与外部培养基分离后，在2 M - H₃PO₄中制备细胞裂解物，以及(ii) 随后通过离子交换高效液相色谱法进行硝酸盐分析，流动相为含有50 mM - H₃PO₄和2%（v/v）四氢呋喃的溶液，用NaOH调节至pH 1.9。当样品中高浓度存在氯离子和硫酸根时，硝酸盐的测定会受到干扰。亚硝酸盐也会产生干扰，但通过用氨基磺酸处理样品可轻松消除。该方法已成功应用于单细胞蓝藻集胞藻硝酸盐转运的研究。
