IAC-SEP® Bisphenol A Immunoaffinity Column Instructions for Use IAC316

IAC-SEP ^® Bisphenol A Immunoaffinity Column

(Product Code: IAC316)

user target audience

The IAC-SEP ^® bisphenol A affinity column can specifically purify bisphenol A in a sample. It uses a columnar agarose gel as a solid phase carrier, and an agarose gel is coupled with a bisphenol A antibody to form an immunosorbent. , packed into columns to make an immunoaffinity column. It is capable of specifically purifying bisphenol A in a sample. Bisphenol A affinity column is widely used in the extraction of beverage, milk and milk powder, urine and other samples. This method is fast, easy to operate and high in accuracy, and plays an important role in improving the quality and safety of food.

principle

The bisphenol A immunoaffinity column can specifically purify the bisphenol A in the sample, mix the sample with the extract, extract, filter, and then pass the filtrate through the bisphenol A affinity column. At this time, the bisphenol A substance is bonded to the antibody in the affinity column. The impurities on the immunoaffinity column were removed with distilled water. The bisphenol A was separated from the antibody by passing through a separation column with methanol. Finally, the filtered methanol solution was injected into HPLC or LC-MS for measurement.

Storage conditions

The affinity column is stored at 2-8 °C and must not be frozen. The shelf life is 18 months. It is recommended to use at room temperature (18-30 ° C).

Experimental solution preparation

pH 7.4 PBS solution: 8.0 g NaCl + 2.90 g Na ₂ HPO4 12H ₂ O + 0.24 g KH ₂ PO4 + 0.2 g KCl plus 990 mL pure water was dissolved, and pure water was added to 1 liter.

Liquid chromatography conditions

Column: Cloversil-C18, 4.6 × 250mm (5um)

Mobile phase: methanol: water = 70:30 (v/v)

Flow rate: 1.0 mL/min

Detector: fluorescence detector, excitation wavelength 227nm, emission wave, 313nm

Injection volume: 20uL

Analysis step

1. < >: Milk : Accurately measure 20 mL of milk sample in a 50 mL centrifuge tube, accurately add 20.0 mL of pure water, mix, and centrifuge at 4000 RPM for 10 min. Accurately remove 20.0 mL of the lower layer of liquid and set aside.

Milk powder: Weigh 5 g (accurate to 0.01 g) of the sample and place it in a 250 mL beaker. 50 mL of water preheated to 50 °C was added to the milk powder and mixed well with a glass rod. If the milk powder is still not completely dissolved, place the beaker in a 50 ° C water bath for 30 min. After dissolving, cool to 20 °C, transfer to a 100 mL volumetric flask, and wash the beaker with a small amount of water. The washing solution is transferred to a volumetric flask, and the volume is adjusted to the mark with water and shaken. Accurately remove 20.0 mL of the lower layer of liquid and set aside.

Infant rice noodles: Weigh 6 g (accurate to 0.01 g) sample and place in a 50 mL centrifuge tube. Accurately add 30.0 mL of 60% acetonitrile water, mix and shake for 30 min. The mixture was then centrifuged at 4000 RPM for 10 min and the upper liquid was filtered through a common filter paper. Accurately measure 10 mL of the filtrate in a 100 mL Erlenmeyer flask, add 50 mL of pure water, dilute and mix, and set aside.

Beverage: Accurately measure 25mL of sample in a 50mL volumetric flask, adjust the pH value to about 7-8, dilute to 50mL with pH7.4 PBS solution , accurately remove 40.0 mL of the lower layer of liquid, and set aside.

1. < > Milk, milk powder (measurement range: 0-50μg/L for milk samples, 0-500μg/kg for milk powder samples)

   The immunoaffinity column was attached to a 20.0 mL glass syringe. Accurately transfer 20.0 mL of sample extract into a glass syringe, connect the air pressure pump to the glass syringe, adjust the pressure so that the solution slowly passes through the immunoaffinity column at a flow rate of about 2 mL/min (1 drop/second) until 2~3 mL of air. Pass through the cylinder. The column was rinsed once with 10.0 mL of water at a flow rate of 2-3 mL/min (1-2 drops/second), all of the effluent was discarded, and 2 mL to 3 mL of air was passed through the column. Accurately add 1.0 mL of chromatographic methanol elution at a flow rate of 1 mL/min to 2 mL/min (1 drop/second). Collect all the eluate in a glass test tube, concentrate and blow dry at 50 °C, and use a mobile phase. Constant volume to 0.5 mL for HPLC or LC-MS analysis.

   Beverage (measurement range: 0-25μg/L)

   The immunoaffinity column was attached to a 50.0 mL glass syringe. Accurately transfer 40.0 mL of sample extract into a glass syringe, connect the air pressure pump to the glass syringe, adjust the pressure so that the solution slowly passes through the immunoaffinity column at a flow rate of about 2 mL/min (1 drop/second) until 2~3 mL of air. Pass through the cylinder. The column was rinsed once with 10.0 mL of water at a flow rate of 2-3 mL/min (1-2 drops/second), all of the effluent was discarded, and 2 mL to 3 mL of air was passed through the column. Accurately add 1.0 mL of chromatographic methanol elution at a flow rate of 1 mL/min to 2 mL/min (1 drop/second). Collect all the eluate in a glass test tube, concentrate and blow dry at 50 °C, and use a mobile phase. Constant volume to 0.5 mL for HPLC or LC-MS analysis.

   Infant rice noodles: (measurement range: 0-100μg/kg)

   The immunoaffinity column was attached to a 50.0 mL glass syringe. Accurately transfer all 60.0 mL of sample extract into a glass syringe, connect the air pressure pump to the glass syringe, adjust the pressure so that the solution slowly passes through the immunoaffinity column at a flow rate of about 2 mL/min (1 drop/second) until 2~3 mL Air passes through the column. The column was rinsed once with 10.0 mL of water at a flow rate of 2-3 mL/min (1-2 drops/second), all of the effluent was discarded, and 2 mL to 3 mL of air was passed through the column. Accurately add 1.0 mL of chromatographic methanol elution at a flow rate of 1 mL/min to 2 mL/min (1 drop/second). Collect all the eluate in a glass test tube, concentrate and blow dry at 50 °C, and use a mobile phase. Constant volume to 0.5 mL for HPLC or LC-MS analysis.

   Note: In order to save the experiment time, the nitrogen blowing step can be omitted, and 1.0 mL of the chromatographic methanol eluent is directly injected into the HPLC test (provided that the liquid chromatograph conditions permit), so the detection sensitivity is reduced by 2 times.

   Precautions

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   C/N number

   article

   21022

   Clover six-stage pumping operating frame (with 1 air pump, 6 10mL syringes)

   20202

   High speed homogenizer with speeds up to 18,000 r/min ~ 22,000 r/min

   31955

   American viam microfiber filter paper (1.5um, 100 sheets / box)

   31240

   Folding fluted filter paper (100 sheets / box), or medium speed qualitative filter paper

   36010

   Disposable plastic beaker (25/pack)

   34000

   Disposable test tube (250 pcs/pack)

   36020

   Plastic funnel (10 pcs/pack)

   35016

   Chromatography pure grade methanol (4 L / bottle)

   C546150-U

   Cloversil C18 Reversed Phase Column 4.6*150mm (5um)

   C546250-U

   Cloversil C18 reversed-phase column 4.6*250mm (5um), data request qq1609095159

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