Ochratoxin 赭麴毒素-定量-快篩試紙-水溶性萃取

• 原理
  側流體免疫層析法 (lateral flow immunochromatographic assay)，為快篩試劑的使用原理，藉由抗體抗原間的專一性與免疫親和力做檢測，被普遍應用在、病毒感染確認、妊娠測試、警局臨檢驗毒等方面。此方法多用於定性檢測，且顯色標定物上也有許多不同的種類，目前大多數的顯色方式是利用抗體吸附在膠體金（colloid gold）來呈色。
  
  Ochratoxin 赭麴毒素，依其在薄層層析法上螢光顏色的不同，通常分成A、B、C三種類型，其中以Ochratoxin A 毒性最強且產量最高，故一般赭麴毒素多以ochratoxin A為代表。赭麴毒素主要是真菌Aspergillus ochraceus、Penicillium verrucosum產生的二次(次級)代謝物。穀類(例如:米、玉米、麥類)、咖啡、香辛植物...等相關加工品或原料可能會受到赭麴毒素的汙染，目前在動物實驗上，已證實赭麴毒素 A 的暴露，會造成腎臟毒性，並引發腎臟癌
   
• 結果判讀
  極限:
  偵測極限(limit of detection, LOD)： 1.1 ppb 
  定量極限(limit of quantification, LOQ)：5 ppb
  偵測範圍:5–25 ppb(樣品超過25ppb需重新稀釋及測量)
• 樣品種類:小麥、米、高粱、玉米和小麥麩皮(For wheat, rice, sorghum, corn and wheat bran)
  
  樣品萃取
  1. Weigh out 10 ± 0.1 g of sample into extraction cup.
  2. Add contents of one (1) MAX 1 aqueous extraction packet to the extraction cup.
  3. Add 50 mL distilled or deionized water to the extraction cup.
  4. Vigorously shake, using hand or mechanical means, for 3 minutes, or blend for 1 minute.
  5. Allow the sample to settle, then filter with a filter syringe or Whatman No. 4 filter paper, collecting a minimum of 3 mL filtrate into a sample collection tube. You may also pipette sample into a 2.0 mL microcentrifuge tube, and centrifuge for 30 seconds using a microcentrifuge.
  
  操作流程
  1. Place the appropriate number of red sample dilution cups and clear sample cups into a sample cup rack. Label cups if necessary.
  2. Add 200 µL of sample diluent (orange-labeled bottle) to each red sample dilution cup.
  3. Add 400 µL of sample extract to the red sample dilution cup containing the sample diluent. Mix by pipetting up and down 5 times.
  4. Fully insert a Reveal Q+ MAX for Ochratoxin test strip into a Raptor cartridge.
  5. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
  a. The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
  b. Scan the QR code found on the tube containing the test strips. The information will be stored on the reader. (The QR code can also be found on the kit Certificate of Analysis.)
  6. Enter the Sample ID if desired.
  7. Select Curve 1 for wheat, rice or sorghum, or Curve 2 for corn and wheat bran samples.
  8. Add 400 µL of sample from the red sample dilution cup to the Raptor cartridge.
  a. The Raptor system will start automatically.
  b. Additional samples can be started in the other ports while the first sample is processing.
  9. Results will be displayed on the Raptor screen after the 5-minute incubation is complete.
  
  稀釋步驟(樣品超過25ppb需重新稀釋及測量)
  1. Add 100 µL of sample filtrate to a sample collection tube.
  2. Add 400 µL of dilution buffer (yellow-labeled bottle) to the sample collection tube. Mix by pipetting up and down 5 times.
  3. Add 200 µL of sample diluent (orange-labeled bottle) to a red sample dilution cup.
  4. Add 400 µL of sample extract (from step 2) to the red sample dilution cup containing the sample diluent. Mix by pipetting up and down 5 times.
  5. Fully insert a Reveal Q+ MAX for Ochratoxin test strip into a Raptor cartridge.
  6. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
  a. The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
  b. Scan the QR code found on the tube containing the test strips. The information will be stored on the reader. (The QR code can also be found on the kit Certificate of Analysis.)
  7. Enter the Sample ID if desired.
  8. Select Curve 1 for wheat, rice or sorghum, or Curve 2 for corn and wheat bran samples.
  9. Add 400 µL of sample from the red sample dilution cup to the Raptor cartridge.
  a. The Raptor system will start automatically.
  b. Additional samples can be started in the other ports while the first sample is processing.
  10. Results will be displayed on the Raptor screen after the 5-minute incubation is complete.
  11. Multiply your result by 5.
   
• 樣品種類:大麥(For barley)
  
  樣品萃取
  1. Weigh out 10 ± 0.1 g of sample into extraction cup.
  2. Add contents of one (1) MAX 1 aqueous extraction packet to the extraction cup.
  3. Add 40 mL distilled or deionized water to the extraction cup.
  4. Vigorously shake, using hand or mechanical means, for 3 minutes, or blend for 1 minute.
  5. Allow the sample to settle, then filter with a filter syringe or Whatman No. 4 filter paper, collecting a minimum of 3 mL filtrate into a sample collection tube. You may also pipette sample into a 2.0 mL microcentrifuge tube, and centrifuge for 30 seconds using a microcentrifuge.
  
  操作流程
  1. Place the appropriate number of red sample dilution cups and clear sample cups into a sample cup rack. Label cups if necessary.
  2. Add 200 µL of sample diluent (orange-labeled bottle) to each red sample dilution cup.
  3. Add 400 µL of sample extract to the red sample dilution cup containing the sample diluent. Mix by pipetting up and down 5 times.
  4. Fully insert a Reveal Q+ MAX for Ochratoxin test strip into a Raptor cartridge.
  5. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
  a.  The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
  b.  Scan the QR code found on the tube containing the test strips. The information will be stored on the reader. (The QR code can also be found on the kit Certificate of Analysis.)
  6. Enter the Sample ID if desired.
  7. Select Curve 1 for barley samples.
  8. Add 400 µL of sample from the red sample dilution cup to the Raptor cartridge.
  a. The Raptor system will start automatically.
  b. Additional samples can be started in the other ports while the first sample  is processing.
  9. Results will be displayed on the Raptor screen after the 5-minute incubation  is complete.
  
  稀釋步驟(樣品超過25ppb需重新稀釋及測量)
  1. Add 100 µL of sample filtrate to a sample collection tube.
  2. Add 400 µL of dilution buffer (yellow-labeled bottle) to the sample collection tube. Mix by pipetting up and down 5 times.
  3. Add 200 µL of sample diluent (orange-labeled bottle) to a red sample dilution cup.
  4. Add 400 µL of sample extract (from step 2) to the red sample dilution cup containing the sample diluent. Mix by pipetting up and down 5 times.
  5. Fully insert a Reveal Q+ MAX for Ochratoxin test strip into a Raptor cartridge.
  6. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
  a. The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
  b. Scan the QR code found on the tube containing the test strips. The information will be stored on the reader. (The QR code can also be found on the kit Certificate of Analysis.)
  7. Enter the Sample ID if desired.
  8. Select Curve 1 for barley samples.
  9. Add 400 µL of sample from the red sample dilution cup to the Raptor cartridge.
  a. The Raptor system will start automatically.
  b. Additional samples can be started in the other ports while the first sample is processing.
  10. Results will be displayed on the Raptor screen after the 5-minute incubation is complete.
  11. Multiply your result by 5.
   
• 樣品:燕麥和綠咖啡(For oats and green coffee)
  
  樣品萃取
  1. Weigh out 10 ± 0.1 g of sample into extraction cup.
  2. Add contents of one (1) MAX 1 aqueous extraction packet to the extraction cup.
  3. Add 40 mL distilled or deionized water to the extraction cup.
  4. Vigorously shake, using hand or mechanical means, for 3 minutes, or blend for  1 minute.
  5. Allow the sample to settle, then filter with a filter syringe or Whatman No. 4 filter paper, collecting a minimum of 3 mL filtrate into a sample collection tube. You may also pipette sample into a 2.0 mL microcentrifuge tube, and centrifuge for 30 seconds using a microcentrifuge.
  
  操作流程
  1. Place the appropriate number of red sample dilution cups and clear sample cups into a sample cup rack. Label cups if necessary.
  2. Add 250 µL of sample diluent (orange-labeled bottle) to each red sample dilution cup.
  3. Add 250 µL of sample extract to the red sample dilution cup containing the sample diluent. Mix by pipetting up and down 5 times.
  4. Fully insert a Reveal Q+ MAX for Ochratoxin test strip into a Raptor cartridge.
  5. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
  a. The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
  b. Scan the QR code found on the tube containing the test strips. The information will be stored on the reader. (The QR code can also be found on the kit Certificate of Analysis.)
  6. Enter the Sample ID if desired.
  7. Select Curve 1 for oats and green coffee samples.
  8. Add 400 µL of sample from the red sample dilution cup to the Raptor cartridge.
  a. The Raptor system will start automatically.
  b. Additional samples can be started in the other ports while the first sample is processing.
  9. Results will be displayed on the Raptor screen after the 5-minute incubation is complete.
  
  稀釋步驟(樣品超過25ppb需重新稀釋及測量)
  1. Add 100 µL of sample filtrate to a sample collection tube.
  2. Add 400 µL of dilution buffer (yellow-labeled bottle) to the sample collection tube. Mix by pipetting up and down 5 times.
  3. Add 250 µL of sample diluent (orange-labeled bottle) to a red sample dilution cup.
  4. Add 250 µL of sample extract (from step 2) to the red sample dilution cup containing the sample diluent. Mix by pipetting up and down 5 times.
  5. Fully insert a Reveal Q+ MAX for Ochratoxin test strip into a Raptor cartridge.
  6. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
  a. The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
  b. Scan the QR code found on the tube containing the test strips. The information will be stored on the reader. (The QR code can also be found on the kit Certificate of Analysis.)
  7. Enter the Sample ID if desired.
  8. Select Curve 1 for oats and green coffee samples.
  9. Add 400 µL of sample from the red sample dilution cup to the Raptor cartridge.
  a. The Raptor system will start automatically.
  b. Additional samples can be started in the other ports while the first sample is processing.
  10. Results will be displayed on the Raptor screen after the 5-minute incubation is complete.
  11. Multiply your result by 5.

• 產品說明書：連結
• 操作手冊：連結
• 物質安全資料表(Safety Data Sheet; SDS)：中文連結、英文連結

參考規範：食品中污染物質及毒素衛生標準

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