|
KMID : 1134820180470020168
|
|
Journal of the Korean Society of Food Science and Nutrition
2018 Volume.47 No. 2 p.168 ~ p.175
|
|
|
Validation of Pepsin-Assisted Extraction and Immunoaffinity-HPLC/DAD Analysis for Vitamin B12 in Seafood
|
|
Mun Geum-Ju
Choi Young-Min
Chun Ji-Yeon
|
|
|
Abstract
|
|
|
|
Vitamin B12 exists in free and bound forms in foods. Use of proteolytic enzymes is recommended for vitamin B12 extraction from protein-rich foods, but information applicable to seafood is very limited. In this study, pepsin-assisted extraction (PE) coupled with immunoaffinity-HPLC/DAD (I/H/D) was validated and applied for determination of total vitamin B12 contents (sum of free and bound) in four types of seafood with different matrices (littleneck clam, blue crab, mackerel, and squid). PE-I/H/D showed good specificity and linearity (R2=0.9992). Limits of detection and quantification of PE-I/H/D were about twice as high (0.015 and 0.106 μg/100 g, respectively) as those of non-PE-I /H/D. Both PE- and non-PE-I/H/D showed good accuracy based on higher than 95% recoveries from the certified reference material, BCR-487. Precision of PE-I/H/D was acceptable based on AOAC guidelines, although it decreased upon application of PE (coefficient variations of repeatability and reproducibility: 9.4 and 13.6% for PE-I/H/D and 6.0 and 10.5% for non-PE-I/H/D, respectively). Free and total vitamin B12 contents of the four types of seafood were analyzed by using the validated PE-I/H/D. The total vitamin B12 contents of littleneck clam (55.16 μg/100 g) and mackerel (5.60 μg/100 g) were significantly higher than corresponding free vitamin B12 contents (46.91 and 4.64 μg/100 g, respectively), indicating that 18∼20% of total vitamin B12 exists in bound form in littleneck crab and mackerel. On the other hand, there was no significant difference between free and total vitamin B12 contents in blue crab and squid within the ranges of 6.09∼6.87 and 2.39∼2.45 μg/100 g, respectively. The results of this study suggest that PE-I/H/D assay would be effective to release vitamin B12 from protein-rich seafood tissues and to produce acceptable selectivity, sensitivity, accuracy, and precision of vitamin B12 analysis, indicating PE-I/H/D is applicable for the determination of vitamin B12 at low levels in seafood.
|
|
|
KEYWORD
|
|
|
vitamin B12, pepsin-assisted extraction, seafood, method validation
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|