biocommaIntroduction
Synthetic colorants often rely on chemicals like benzene,toluene, and naphthalene as key raw materials. They’re favoredfor their low cost, vibrant colors, strong pigmentation, and widerange of shades. However, excessive consumption can posehealth risks. Biocomma utilizes the polyamide SPE cartridges,developing a HPLC method with simplified operation and goodrecovery to detect tartrazine, sunset yellow, new red, carminecochineal, amaranthus red, allura red, and brilliant blue infoods for your reference.
Experiment
Preparation
Prepare ethanol-ammonia solution: add 700 mL of ethanol and4 mL of ammonia. Dilute to 1 L by water
Extraction of juice and beverage sample
Weigh 2.0 g of the sample (accurate to 0.001 g) into a 50 mLcentrifuge tube. Add 25 mL of ethanol-ammonia solution, andvortex for 5 min. Extract by ultrasonic treatment for 15 minat 50 °C, and centrifuge at 8000 r/min for 5 min. Transfer thesupernatant into a clean 50 mL centrifuge tube. Add 15 mL ofthe ethanol-ammonia solution to the remaining sample. Repeatthe extraction procedures. Combine the supernatants anddilute to 50 mL. Pipette 10 mL of supernatant to a clean tubeand concentrate to 2 mL at 50°C. Add 10 mL of 5% aqueousmethanol solution and mix well. Adjust the pH to 3~4 by 20%citric acid. The sample is ready for purification.
Extraction of ham, potato chips, bread and chicken wingssample
Weigh 2 g of the sample into a 50 mL centrifuge tube. Add 25mL of ethanol-ammonia solution, and vortex for 5 min. Extractby ultrasonic treatment for 15 min at 50 °C, and centrifuge at8000 r/min for 5 min. Transfer the supernatant into a clean 50mL centrifuge tube. Add 15 mL of the ethanol-ammonia solutionto the remaining sample. Repeat the extraction procedures.Combine the supernatants and dilute to 50 mL. Pipette 10mL of supernatant to a clean tube and concentrate to 2 mL at50°C. Add 10 mL of 5% aqueous methanol solution and mixwell. Adjust the pH to 3~4 by 20% citric acid. The sample isready for purification.
Purification (Copure® Polyamide (PA) SPE Cartridges, 500mg/ 6 mL)
Activation: activated the cartridge by 6 mL of methanol, then 6mL of water.
Loading: add prepared sample to the activated cartridge.
Washing: add 5 mL of water, then 5 mL of methanol. Drain thecartridge.
Elution: add 7 mL of 5% ammoniated methanol solution andcollect the eluate. Evaporate to about 0.3 mL at 45 °C. Diluteto 2 mL with 0.02 mmol/L ammonium acetate (pH=9.0). Vortexto mix well. After filter by PTFE membrane, the sample is readyfor LC-MS/MS analysis.
Instrumental Conditions
Instrument: liquid chromatograph, ThermoFisher U3000
Chromatographic column: Agilent ZORBAX SB-C18 (4.6mm×250 mm, 5 μm)
Mobile phase: A: 0.02 mol/L ammonium acetate solution, B:methanol
Flow rate: 1.0 mL/min
Column temperature: 30 ℃
Injection volume: 10 μL
Detector: UV detector
Detector wavelength range: 400~800 nm. (Tartrazine: 415 nm.New red, carmine cochineal, amaranthus red, sunset yellow,allura red and erythrosine: 520 nm. Brilliant blue: 630 nm.)Table 1. Gradient Elution Program
Results
Table 2. Spiked Recovery Rate
Ordering Information
