Page 45 - Application Notebook - Solution for Food Development

LAAN-A-LM-E103

Application Liquid Chromatography Mass Spectrometry

News Comprehensive Analysis of Primary and Secondary Metabolites

in Citrus Fruits Using an Automated Method Changeover
No.C132 UHPLC System and LC/MS/MS System [LCMS-8050]

Due to an increasing trend for healthy lifestyles, the In food analysis, it is very important to carry out
functional properties of agricultural products and foods comprehensive analysis of primary and secondary
are being emphasized alongside their taste qualities, metabolites that contribute to the color, smell, taste,
and the production and development of foods with and functional properties of food. To date, there has
high additional value is becoming increasingly been only a few examples of performing these analyses
important. Metabolome analysis, which is the simultaneously.
comprehensive analysis of all metabolites present in a In this article, we describe a method of performing
living organism, is now being applied in a variety of comprehensive analysis of primary and secondary
areas and not just to analyze the metabolites of living metabolites in food (major organic acids, amino acids,
organisms. Metabolome analysis is becoming an sugars, carotenoids, and flavonoids) using LC/MS/MS,
important tool in the food sector in areas such as food and also present an example application of this
processing and plant breeding. analytical method using citrus fruits as an actual
sample.

n Sample Preparation
We used seven citrus fruits (mikan orange, ponkan, shiranui orange, amakusa orange, hassaku orange, buntan
orange, and hyuganatsu) shown in Fig. 1. Each metabolite was fractionated and extracted using two solvents, as
shown in Fig. 2.

Mikan Trovita Homogenize the flesh and peel of each citrus fruit
(C. unshu) Orange (separate flesh and peel in advance)
Ponkan Weigh out 10 g of flesh and 5 g of peel
Kiyomi
(C. reticulata)
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orange Add 20 mL of water/ethanol Add 20 mL of hexane
Shiranui
(C. reticulate shiranui)
Amakusa
Centrifugation step
True cultivars
Hybrid cultivars Filtration step

Buntan
??? Yuzu
(C. maxima) Dilution Dilution Dilution
(1/1000) (1/10000) (1/5)
? Organic acids Sugars Carotenoids
Hassaku Amino acids
(C. hassaku) Flavonoids
Hyuganatsu
(C. tamurana)
Fig. 2 Sample Pretreatment Protocol

Fig. 1 Taxonomic Tree of Seven Citrus Fruits