虾中吲哚的高压液相色谱法：方法的建立与协同研究

High pressure liquid chromatographic method for indole in shrimp: development of method and collaborative study.

作者信息

Chambers T L, Staruszkiewicz W F, Bohm G, Bond J F, Carr R, Edge D J, Everett R L, Illuminati J C, La Rose J, McMurtrey K, Miller G, Panaro K W, Smith B H

出版信息

J Assoc Off Anal Chem. 1981 May;64(3):592-602.

PMID:7240065

Abstract

A collaborative study on the determination of indole in shrimp was conducted in which a high pressure liquid chromatographic (HPLC) method and a spectrofluorometric method were compared with the AOAC gas-liquid chromatographic (GLC) method (18.075-18.078, 13th ed.). In the HPLC method, 10 g shrimp was blended with methanol, an internal standard was added, and the extract was filtered. Indole was separated on an octadecylsilane reverse phase column, using 60% MeOH-H2O, and quantitated with a fluorescence detector (excitation 280 nm, emission 330 nm) by comparing the indole peak height with that of an internal standard, 2-methyl-indole. Recoveries at a 25 micrograms/100 g level averaged 104% with a range of 90-127%, and at a level of 35 micrograms/100 g averaged 102% with a range of 93-112%. In the spectrofluorometric method, 25 g shrimp was extracted with 2% EtOAc-hexane. After several washes, indole was partitioned into a saturated NaCl-MeOH solution and its fluorescence was measured (excitation 280 nm, emission 332 nm). Recoveries at a 25 micrograms/100 g level averaged 93% with a range of 0-255% and at a level of 35 micrograms/100 g averaged 64% with a range of 0-107%. Recoveries obtained by the AOAC-GLC method at a level of 25 micrograms/100 g averaged 96% with a range of 81-116% and at a level of 35 micrograms/100 g averaged 101% with a range of 81-119%. The coefficients of variation were 20, 10, and 64% at a 25 micrograms/100 g level for the GLC method, the HPLC method, and the spectrofluorometric method, respectively. The HPLC method was adopted as official first action for indole levels in shrimp exceeding 1 microgram/100 g.

摘要

开展了一项关于测定虾中吲哚的合作研究，将高压液相色谱法（HPLC）和荧光分光光度法与美国官方分析化学师协会（AOAC）的气液色谱法（GLC）（第13版，18.075 - 18.078）进行比较。在HPLC法中，将10克虾与甲醇混合，加入内标，然后将提取物过滤。吲哚在十八烷基硅烷反相柱上分离，使用60%甲醇 - 水，并用荧光检测器（激发波长280纳米，发射波长330纳米）通过比较吲哚峰高与内标2 - 甲基吲哚的峰高进行定量。在25微克/100克水平下回收率平均为104%，范围为90 - 127%；在35微克/100克水平下平均为102%，范围为93 - 112%。在荧光分光光度法中，用2%乙酸乙酯 - 己烷提取25克虾。经过几次洗涤后，将吲哚分配到饱和氯化钠 - 甲醇溶液中并测量其荧光（激发波长280纳米，发射波长332纳米）。在25微克/100克水平下回收率平均为93%，范围为0 - 255%；在35微克/100克水平下平均为64%，范围为0 - 107%。AOAC - GLC法在25微克/100克水平下回收率平均为96%，范围为81 - 116%；在35微克/100克水平下平均为101%，范围为81 - 119%。在25微克/100克水平下，GLC法、HPLC法和荧光分光光度法的变异系数分别为20%、10%和64%。对于虾中吲哚含量超过1微克/100克的情况，HPLC法被采用为官方首次行动方法。