Yoshida A, Takagaki Y, Nishimune T
Osaka Prefectural Institute of Public Health, Japan.
J AOAC Int. 1995 May-Jun;78(3):807-11.
A competitive enzyme immunoassay (EIA) for carminic acid was investigated. Monoclonal anticarminic acid antibody was obtained from A/J mice immunized with carminic acid-human immunoglobulin G (IgG) conjugate. Carminic acid was extracted with distilled water from beverage, jelly, candy, pasta sauce, yogurt, or ice cream samples. Ham or fish paste samples were digested with pronase, then carminic acid was extracted from samples with sodium hydroxide solution. The extract was diluted more than 10-fold with 1% gelatin in borate buffer solution. Microtiter plates were coated with carminic acid-bovine serum albumin (BSA) conjugate or just BSA. Goat anti-mouse IgG(H+L)-peroxidase complex was used as a second antibody, and 3,3',5,5'-tetramethylbenzidine was used as a substrate for the peroxidase. The working range for quantitative analysis was 0.3-10 ng/mL, and the detection limit was 0.2 micrograms/g original sample. Recoveries of carminic acid by this assay were > 95% for milk beverage and jelly, and > 85% for yogurt and fish paste. Carminic acid was detected in 7 of 26 red-colored commercial food products and ranged from 3.5 to 356 micrograms/g. This EIA system also responded to the structural analogue of carminic acid, laccaic acid.
对胭脂红酸的竞争性酶免疫测定法（EIA）进行了研究。用胭脂红酸-人免疫球蛋白G（IgG）偶联物免疫A/J小鼠，获得了抗胭脂红酸单克隆抗体。从饮料、果冻、糖果、意大利面酱、酸奶或冰淇淋样品中用蒸馏水提取胭脂红酸。火腿或鱼酱样品用链霉蛋白酶消化，然后用氢氧化钠溶液从样品中提取胭脂红酸。提取物在硼酸盐缓冲溶液中用1%明胶稀释10倍以上。微量滴定板用胭脂红酸-牛血清白蛋白（BSA）偶联物或仅用BSA包被。山羊抗小鼠IgG（H+L）-过氧化物酶复合物用作二抗，3,3',5,5'-四甲基联苯胺用作过氧化物酶的底物。定量分析的工作范围为0.3-10 ng/mL，检测限为0.2微克/克原始样品。该测定法对牛奶饮料和果冻中胭脂红酸的回收率>95%，对酸奶和鱼酱的回收率>85%。在26种红色商业食品中有7种检测到了胭脂红酸，含量范围为3.5至356微克/克。该EIA系统也对胭脂红酸的结构类似物紫胶酸有反应。
