Tribo TM Ochratoxin A Rapid Test Strip User Guide (Catalog No. TBS11161) TribioScience, Inc.
General Description Ochratoxin A (OTA) is a toxic secondary metabolite produced by some Aspergillus and Penicillium species, and is widely present in grains (such as cereals), spices, dried fruit and coffee beans. Ochratoxin A can be accumulated in the meat of animals, and therefore can be found in meat and meat products. It possesses acute liver and kidney toxicity, and is a teratogen, mutagen, and carcinogen, and poses serious health risks to human. Tribo TM ochratoxin A rapid test strip provides a rapid and convenient test for OTA with a colloidal gold immunochromatographic design. It provides a fast (results shown in 10 minutes), simple (no instrument is required, and results can be observed visually), sensitive and reliable detection approach for the presence of ochratoxin A in grains and grain products, spices, and coffee beans. The lower limit of detection (LOD) of ochratoxin A in the sample is 2 ng/ml (2 ppb). Intended Use The Tribo TM ochratoxin A test strip is a lateral flow strip test for rapid detection of ochratoxin A in grain samples. Assay sensitivity: 2 ng/ml (2 ppb) Safety Instructions To receive complete safety information on this product, contact AccuAffinity, Inc. and request Material Safety Data Sheet. Assay Principles Tribo TM ochratoxin A test strip is based on the principle of colloidal gold immunochromatography. An anti-ochratoxin A antibody is conjugated to colloidal gold and placed on conjugate pad. Colloidal gold provides red color to visualize antibody-antigen binding. Ochratoxin A antigen is immobilized on nitrocellulose membrane. After test sample is loaded onto sample pad, it mixes with gold-antibody conjugate and migrates together along the membrane. If sample contains no ochratoxin A, antibody conjugated to colloidal gold will bind the antigen immobilized on membrane, leading to clear red color presented on membrane detection line where the test antigen is immobilized (indicating a negative result). If ochratoxin A is present the test sample, it will bind gold-antibody conjugate and prevent its binding onto the antigen line on membrane. As a result, no color will be visible on detection line on membrane (indicating a positive result). - 2 -
Reagents and Materials in each pack a) 1 ochratoxin A test strip b) 1 disposable dropper c) 1 pack of desiccant d) 1 bottle of sample diluent Sample Collection and Test Procedure Equilibrate test strip to room temperature (20-25 o C) Equilibrate test sample to room temperature (20-25 o C) a) Pulverize over 5 g of representative grain sample, and pass through a sieve with mesh size of 20. Place 2 grams of this pulverized sample into a 50 ml centrifuge tube. b) Add 8 ml ethyl acetate (EtAc) into this centrifuge tube and close lid tightly. Shake violently (or vortex) for 5 minutes and let stand afterwards. If less than 3 ml supernatant is present after sedimentation, centrifuge at 4,000 rpm for 1 minute. c) Transfer 3 ml supernatant into a clean glass beaker, blow air over surface until solvent evaporates completely. Re-dissolve residue in 200 µl sample diluent (included with the test strip) by shaking and mixing thoroughly. Take 100 µl of this re-dissolved solution and add another 100 µl sample diluent to obtain test solution. d) Add 2 drops of test solution described above into sample loading well with a disposable dropper. e) Observe result in 5-10 minutes. Result Interpretation Test result is interpreted by observing test line and control line shown in result window. Negative (-): both test and control lines are present, indicating ochratoxin A concentration in sample is lower than 2 ng/ml; Positive (+): control line is present, and test line is absent. This result indicates ochratoxin A centration is higher than 2 ng/ml in the sample; Invalid test: no control line is present. Please repeat the test using a new test strip following instructions on this user guide. - 3 -
Negative Positive Invalid Sample Loading Sample Loading Sample Loading Test Line Test Line Test Line Control Line Control Line Control Line Precautions 1. Test strip is for one-time use only. Please use the test strip on the same day the package is opened. 2. Do not use tap water, purified water, or distilled water as negative control. 3. Test should be performed at room temperature, and test strip and sample both need to be equilibrated to room temperature before the test. 4. If no liquid movement is observed in the test window 30 seconds after test solution is added to sample loading well, add one more drop of test solution. 5. Positive samples identified by test strip are recommended to be verified with other detection methods (such as HPLC or GC-MS). Storage and Expiration Date Storage: Store in a cool (room temperature) and dry place in intact original packaging. Expiration Date: 12 months after manufacturing date. Technical Assistance For ordering or technical assistance regarding this product, or for additional information about TribioScience products, please email info@tribioscience.com or call (650) 917-9269. General Limited Warranty - 4 -
TribioScience, Inc. warrants its manufactured products against defects in materials and workmanship when used in accordance with the applicable instructions for a period not to extend beyond a product s printed expiration date. TribioScience makes no other warranty, expressed or implied. There is no warranty of merchantability or fitness for a particular purpose. The warranty provided herein and the data, specifications and descriptions of TribioScience products appearing in published catalogues and product literature may not be altered except by express written agreement signed by an officer of TribioScience. Representations, oral or written, which are inconsistent with this warranty or such publications are not authorized and, if given, should not be relied upon. In the event of a breach of the foregoing warranty, TribioScience Inc. s sole obligation shall be to repair or replace, at its option, any product or part thereof that proves defective in materials or workmanship within the warranty period, provided the customer notifies TribioScience promptly of any such defect. The exclusive remedy provided herein shall not be deemed to have failed of its essential purpose so long as TribioScience is willing and able to repair or replace any nonconforming TribioScience product or part. TribioScience shall not be liable for consequential, incidental, special or any other indirect damages resulting from economic loss or property damage sustained by a customer from the use of its products. However, in some states the purchaser may have rights under state law in addition to those provided by this warranty. For research use only. - 5 -
Tribo TM 赭曲霉毒素 A 快速检测试纸卡使用说明书 ( 产品编号 :TBS11161) 1. 产品概述 : 赭曲霉毒素 A(OTA) 是由曲霉菌和青霉属菌株产生的一种有毒真菌次级代谢产物, 广泛存在于谷物及谷物产品 香料和咖啡豆等产品中 赭曲霉毒素 A 具有很强的肝脏和肾脏毒性, 并有致畸 致突变和致癌作用, 严重危害人类健康 赭曲霉毒素一般用薄层色谱 高效液相色谱法检测, 但因其操作繁杂 费用昂贵而影响实际应用 赭曲霉毒素 A 快速检测试纸卡将胶体金免疫层析原理应用于赭曲霉毒素 A 含量的快速测定, 用于测定谷物及谷物产品 香料和咖啡豆等产品中赭曲霉毒素 A 的存在 本品具有灵敏度高 特异性好, 并且操作简便的特性, 不需要任何仪器设备, 可通过肉眼 直接判读结果 样本中赭曲霉毒素 A 的最低检出限为 2ng/ml(2ppb) 2. 用途及灵敏度 : 本产品赭曲霉毒素 A 快速检测试纸卡适用于测定谷物及谷物产品 香料和咖啡豆等产品中 赭曲霉毒素 A 的存在 检测灵敏度 :2ng/ml(2ppb) 3. 检测原理 : 赭曲霉毒素 A 快速检测试纸卡将胶体金免疫层析原理应用于赭曲霉毒素 A 含量的快速测定 在试纸卡中, 抗赭曲霉毒素 A 抗体偶联到胶体金颗粒上, 赭曲霉毒素 A 测试抗原固定在纤维素滤膜上 如果样品不含有赭曲霉毒素 A, 胶体金抗体偶联物与固定抗原结合, 在膜上显示检测线, 检测结果为阴性 如果样品中含有赭曲霉毒素 A, 赭曲霉毒素 A 结合在胶体金抗体偶联物上, 抑制了胶体金抗体偶联物与固定抗原的结合, 不显示检测线, 检测结果为阳性 4. 试纸卡产品组成 : 试纸卡 (1 块 ); 一次性吸管 (1 支 ); 干燥剂 (1 块 ); 样品稀释液 (1 瓶 ) 5. 样品处理 : 1. 取 5g 以上有代表性的样品粉碎 ( 过 20 目筛 ), 准确称取 2g 均匀粉碎试样加入到 - 6 -
离心管中 2. 向离心管中准确加入 8mL 乙酸乙酯, 将瓶塞盖紧密封, 用力振荡 5 分钟 ( 如上清不足 3ml,4000rpm 离心 1 分钟 ) 3. 用吸管取 3mL 上清液到小玻璃杯中, 吹干滤液, 用 200μl 稀释液复溶, 振荡混匀 ; 取 100μl 复溶液再加入 100μl 稀释液混匀, 待测 6. 检测步骤 用一次性吸管垂直滴加 2 滴样本于加样孔内, 开始计时,5-10 分钟判读结果 7. 结果判定 : 阴性 (-): 检测线 (T) 和控制线 (C) 都显色,T 线深度等于或深于 C 线 表明样品中赭曲霉毒素 A 的浓度低于 2ng/ml 阳性 (+): 检测线 (T) 无色, 控制线 (C) 显色,T 线不显色或者浅于 C 线深度 表明样品 中赭曲霉毒素 A 的浓度高于 2ng/ml 无效 : 控制线 (C) 无色, 表示使用方法不正确或试纸卡失效, 结果无效 请对照使用说明书, 用新的试纸卡重新测定 阴性阳性检测无效 加样孔加样孔加样孔 T 检测线 T 检测线 T 检测线 C 控制线 C 控制线 C 控制线 8. 注意事项 : (1) 赭曲霉毒素 A 快速检测试纸卡为一次性使用 打开包装后, 请在当日使用 (2) 请勿使用自来水 纯化水及蒸馏水作为阴性对照 (3) 在室温 (20-25 o C) 下使用, 使用前将检测卡和待检样本恢复至室温 (20-25 o C) 测试 (4) 如滴加检测液后 30 秒内, 在测试窗口无液体移行, 则再补加 1 滴检测液 (5) 本品检测为阳性的样本, 建议采用更精确的方法 ( 如 HPLC 或 GC-MS) 加以复核 9. 储存条件和有效期 : 赭曲霉毒素 A 快速检测试纸卡原包装在室温 (20-25 o C) 下储存, 置于阴凉避光干燥处 有效期为 12 个月, 产品批号及有效期见包装袋 本产品仅供研究使用 - 7 -