Abstract
Objective: The aim of the present study was to validate HPLC method for analysis bromelain levels in pineapple water and application that method to determine the storage time of pineapple water. Methods: The reversed phase of HPLC method was tested and optimized before it is validated. The composition and the flow rate of the mobile phase is the optimized parameter. The analytical parameters validated were detection limits, linearity, accuracy and precision. Pineapple water was stored for 8 h at 10ºC and bromelain was determined using the validated HPLC method. Result: The optimum mobile phase composition was methanolwater (70:30) with a flow rate of 1 ml/min. The detector concentration-response was linear with coefficient of determination of 0.993. The accuracy of HPLC method at a recovery of 1 and 2% bromelain was 106.37 ± 1.94% and 98.12 ± 1.29% (n = 5), respectively. The precision, expressed as the coefficients of variation (CV), at 1 and 2% bromelain were 1.83 and 1.32% (n = 5), respectively. Bromelain level at zero time was 81.53%. After storage for 8 h at 10ºC, bromelain levels in pineapple juice appeared to decrease not statistically significant (p > 0.05), with to mean value of 78.46 ± 2.88%. Conclusion: The HPLC method developed was valid to analyze accurately concentrations of bromelain in pineapple water and it can be used to study the shelf life of pineapple water based on bromelain content. The bromelain content in pineapple juice was not statistically significantly different after 8 h storage at 10°C.
Highlights
Pineapple water was stored for 8 h at 10oC and bromelain was determined using the validated high performance liquid chromatography (HPLC) method
The HPLC method developed was valid to analyze accurately concentrations of bromelain in pineapple water and it can be used to study the shelf life of pineapple water based on bromelain content
Bromelain can be used as an indicator of the quality of pineapple fruit, because bromelain is the largest component of the protease enzyme in pineapple.[2]
Summary
Base on vitamin C level, shelf life studies of pineapple juice have been reported.[7,8] Bromelain can be used as an indicator of the quality of pineapple fruit, because bromelain is the largest component of the protease enzyme in pineapple.[2]. Bromelain is sensitive to high temperatures, acidity and organic solvents.[10,11,12] This instability will result in decreased enzymatic activity and limit its pharmacological effects.[2] a study is needed to determine the bromelain content during storage of pineapple fruit water. Validation of High-Performance Liquid Chromatography for Determination of Bromelain in Pineapple (Ananas comosus (L) Merr) Water. Et al.: Validation of High-Performance Liquid Chromatography for Determination of Bromelain in Pineapple (Ananas comosus (L) Merr) Water. The validated method is used to determine the effect of storage of pineapple fruit water on levels of bromelain
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