1. Multi-functional purification column + KRC photochemical post-column derivatization method
1 Introduction
Multi-functional purification column is composed of a variety of solid-phase composite fillers, including not only macromolecular substances, but more importantly, a variety of active lipophilic (non-polar) and charged (polar) substances, used to adsorb various esters, leaves Interferors such as flavin, chemicals, carbohydrates and proteins. Multi-functional purification column + KRC derivatization method is to use PRIBOLAB's Pribofast 226/260 multi-functional purification column to separate aflatoxins B1, B2, G1 and G2 well, and then use HPLC/FLD+KRC post-column derivatization system. The content of aflatoxin B1, B2, G1, G2 can be effectively determined.
2 Multi-functional purification column features:
2.1 Multi-functional purification column can simplify the sample extraction and purification steps without cleaning and elution.
2.2 Can achieve a variety of toxin detection, effectively reducing the cost of testing;
2.3 HPLC+FLD+KRC derivatization can effectively improve the detection sensitivity and achieve a lower detection limit (aflatoxin B1 0.05ppb);
2.4 Photochemical derivatization is easy to operate, no need to use derivatization reagents (usually, derivatization reagents are needed after electrochemical column derivatization, the analysis cost is high, the operation is troublesome, and the instrument failure rate is high);
2.5 One-step purification, the recovery rate is as high as 90%.
2.6 Widely applicable to all kinds of agricultural products: grain, starch, animal feed, food, nuts, cottonseed, etc.
3 Multi-function purification column / KRC derivative method steps:
3.1 Extraction of samples: grinding and pulverizing, weighing 25g samples; mixing samples with acetonitrile/water (84/16 V/V),
High speed homogenization for 1 minute.
3.2 Filtration: The extracted sample quantitative filter paper is filtered to collect the filtrate;
3.3 Take 4-6ml of filtrate quickly through the multi-functional purification column PribofastM226 or Pribofast M260
3.4 The column solution was collected and used directly for HPLC detection. The concentration of aflatoxin was directly recorded after KRC derivatization.
Second, immunoaffinity column + KRC photochemical post-column derivatization
1 Introduction
The Pribofast aflatoxin total immunoaffinity column selectively adsorbs toxins from the sample solution, thereby providing a very targeted purification of the toxin sample, and then effectively determining the Aspergillus flavus using the HPLC/FLD+KRC post-column derivatization system. The concentrations of toxins B1, B2, G1, and G2 increase the accuracy and sensitivity of the assay.
2 Immunoaffinity column / KRC derivative characteristics:
2.1 The specificity is very strong, which makes the sample purification effect excellent;
2.2 Using the immunoaffinity column operating frame can make the sample clean quickly and improve the detection efficiency.
2.3 HPLC+FLD+KRC post-column derivatization, effectively improve the detection sensitivity and achieve a lower detection limit (aflatoxin B1 0.05ppb).
2.4 Derivatization is convenient, and photochemical derivatization does not require the use of derivatization reagents (usually derivatization is required after electrochemical column derivatization, the analysis cost is high, the operation is troublesome, and the instrument failure rate is high).
2.5 High recovery rate, reaching over 90%
3 Introduction of immunoaffinity column method:
3.1 Extraction of sample: grinding, weighing 50g sample; sample with methanol / water (80 / 20 V / V) mixture
Mix at high speed for 1 minute.
3.2 Filtration: The extracted sample filter is passed through the glass fiber filter paper.
3.3 Install the regenerative immunoaffinity column in the pump flow frame for column purification: after the Pribofast aflatoxin total immunoaffinity column (IAC-010-3) activation, elution, elution three steps, elution The solution can be directly used for HPLC detection;
3.4 Determination: direct injection of HPLC, direct recording of aflatoxin concentration after column and KRC photochemical column derivatization;
Third, the total amount of aflatoxin detected by enzyme-linked immunosorbent assay
1 A direct competition ELISA method is used to determine the total amount of aflatoxin in the sample by measuring the absorbance of the sample and the sample of the standard using a microplate reader.
2 kit detection time: 15 minutes + 15 minutes
3 Detection limit: 0.1ppb
4 Cross-reaction rate:
Aflatoxin B1
100%
Aflatoxin B2
80%
Aflatoxin G1
101%
Aflatoxin G2
100%
AflatoxinM1
25%
About Pribofast KRC Post-column Derivatives and HPLC Parameters
â—Pribofast KRC post-column photochemical derivative wavelength: 254 nm
â—HPLC-column: Aflatoxin-specific column 150 x 4.6 mm; C-18
â— Mobile phase: methanol / water (45 / 55 v / v)
◠Flow rate: 1.0 mL/min Column temperature: 30 °C
◠Injection volume: 20μL
Fluorescence detector: λ-excitation wavelength: 365 nm λ-emission wavelength: 440 nm
â—High performance liquid chromatograph needs to be prepared: PriboLab Pribofast KRC post-column photochemical derivatizer with 1ml reaction cell, nano tube 254nm
Special column for aflatoxin:
product number
Product name and specifications
Quantity
IAC-010-3
Pribofast, Aflatoxins B1, B2, G1, G2 25 columns/pck 3mL, widebore column
Aflatoxin total immunoaffinity column 3ml, 25 / box
1 box
M226
PriboFast®226 Multifunctional cleanup column for Aflatoxin and zearalenone
Mycotoxin multi-purpose purification column for aflatoxin zearalenone detection 25 / box
1 box
PriboFast® 260
PriboFast® 260 Multifunctional cleanup column for Aflatoxin and zearalenone
Mycotoxin multi-purpose purification column for aflatoxin zearalenone detection 25 / box
1 box
PR K010-96
Pribofast ELISA Kit (96-well plate/stripes) for detection of Aflatoxin B1, B2, G1, G2 in food and feed samples
Aflatoxin total ELISA kit, 96 wells
1 box
STD#1081
Standard, Solution of 4 different aflatoxins in acetonitrile B1, G1
Aflatoxin B1B2G1G2 mixed standard. 1ml, 4 degrees dark storage
Four separate solids standards for aflatoxin B1B2G1G2 are also available.
1 box
KRC-25
Photochemical Reactor for Enhanced Detection Aflatoxin B1, B2, G1, G2; Pribofast KRC-25 230V 50 HzUVE 254 nm lamp
Pribofast KRC photochemical post-column derivatization system, 254nm nano UV lamp
1 set
ODS-150
HPLC-Column for Aflatoxin Analysis, C18 150 x 4.6 mm Aflatoxin-specific LC column
1
Pump Flow Operator / Immunoaffinity Column Operator for Immunoaffinity Column Sample Purification
(including air pump adapter and other accessories, multi-purpose connector is optional)
1 set
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