A Simple, Robust and Handheld Sample Preparation Technique for Quantifying Canthaxanthin in Chicken Liver, Fat and Egg Yolk

The present chapter described a fast, simple, and small-scale method of sample preparation followed by HPLC coupled with a photodiode array detector (PDAD) for quantification of canthaxanthin in chicken liver and fat, egg yolk. The permitted Canthaxanthin (CX) is a red pigment and is commonly added to feeds for chickens in order to achieve the desired chicken product color. The HPLC-PDAD was performed on a C18 column with an isocratic mobile phase. The analyte was extracted from the sample using a handheld ultrasonic homogenizer and purified by MonoSpin®-SI, a centrifugal monolithic silica spin mini-columns, and quantified < 15 min. The system-suitability evaluation is an essential parameter of HPLC determination, and it ascertains the strictness of the system used. The suitability was evaluated as the relative standard deviations of peak areas and retention times calculated for 20 replicate injections of a spiked egg yolk sample. The proposed method obtained average recoveries for the analyte analyte in the range of 93.5 - 101.0% with relative standard deviations ≤ 2.7%. The Limits of quantification in the liver, fat, and egg yolk were 0.48, 0.47, and 0.5 µg/g, respectively. The present procedure provided an easy-to-use, rapid, and space-saving and resulted in high recovery and repeatability with considerable saving of analysis time/cost. The procedure may be proposed as an international harmonized method for determining CX in domestic/imported chicken products.