SPL-M100 Application Note

No. AS200602EN

Compositional Comparison of Beers by On-Line Solid Phase
Derivatization SPE-GC/MSIntroduction

In conventional GC/MS analysis of metabolomics, extraction, lyophilization, and derivatization werecomplicated and time-consuming, and the data varied from person to person. To solve this problem,SPL-M100for SPE-GC system

we have dramatically shortened the time required, simplified the process, and achieved high accuracy
with our proprietary “solid-phase derivatization method”. Amino acids and organic acids are held inthe solid phase by ion-exchange interactions, and then passed through acetonitrile to dehydrate andSample

wash the solid phase. In this study, we attempted to compare the composition of beers using theSPL-M100 system, which fully automates these processes.

SamplePretreatment flow

A. Beer

Sample 50 µL

Malt content of more than

Add Water 150 µL

50% and less than 5% of

Add ACN 800 µL

secondary ingredients

than 5% of the secondary

Automation
pretreatment：15 minFlash-SPE ACXs

Conditioning
ACN-Water(1/1)
ACN

Load[Retention] Extract 50 µL

Centrifuge 14000 rpm, 3 minMalt less than 50% or more

Beer

SPE-GC：SPL-M100Vials

Shaking 37 ℃, 5 min

B. Third-category beer

On-line solid-phase derivatization

Washing ACN-Water(1/1)

Add 0.1N NaOH

Dewater ACN

ingredients, plus spirits

Extract

derived from wheat, or madeInformation【Sample】
・Beer
・Third-categorybeer・Non-alcoholic

Infiltration Methoxyamine / Pyridinefrom ingredients other than

SPD Methoximation, 3 min

wheat or malt

Infiltration MSTFA-Toluene(3/1)

C. Non-alcoholic

SPD TMS, 1 min

Alcohol content less than 1%.

Elution Hexane【Target】
・Amino acids
・Amines・Nucleobases,
・organic acids

GC-MS Sprit 1:50

Analytical condition
PTV Injector
LVI-S250 (AiSTI Science)
Insert Type
Spiral Insert
Injector Temp. 220℃(0.5min)-50℃/min-290℃(16min)
GC-MS
Inlet Mode
Split 1:50
Flow Mode
Constant Flow, 1 ml/min
Pre-Column
0.25mm i.d. x 1m
ColumnVf-5ms, 0.25mm i.d. x 30m, df;0.25µm
Oven Temp.
100℃(2min)-10℃/min-220℃-30℃/min-320℃
Trans. Temp. 290℃
MS Method
SCAN, m/z;70-470

SPE-GC-MS system for metabolome analysis
SPL-M100 / GCMS-TQ8040NXAiSTI SCIENCE
Product

SPE-GC interface
38. Tyrosine-3TMS

SPL-M100
45. Gluconic acid-6TMS

33. Citric acid-4TMS

28. Phenylalanine-2TMS

23. GABA-3TMS

18. Malic acid-3TMS

11. Proline-2TMS

14. Succinic acid-2TMS

6. Valine-2TMS

2. Alanine-2TMS

Comparison of SCAN total ion chromatograms for each beer by this method

Solid-phase cartridge

Flash-SPEGC Lage volume
injection

LVI-S250

A. Beer

B. Third-category beer
AiSTI SCIENCE CO.,Ltd.

C. Non-alcoholic

Tel：+81-73-475-0033
as@aisti.co.jp

E-mail：as@aisti.co.jp
www.aisti.co.jp

HP：www.aisti.co.jp

AiSTI Application Note
■Comparison of the amount of ingredients in each beer category (Beer, Third-category beer, Non-alcoholic)【Result and Discussion】
The RSD. (reproducibility) of each component
was all less than 10%, confirming high accuracy.

■Reproducibility of peak area values for beer using this method (n=5) The TIC (total ion chromatogram) shows goodpeak shape and separation, indicating that thewashingandextraction

dehydration

functionallybywork

solid-phase
and

that

derivatization is efficiently performed in thesolid phase. This method is considered to be ahighly accurate analytical method quantitatively
after solid-phase extraction.
The solid-phase derivatization of this analyticalmethodsimplifies

theextraction

process,

improves accuracy, and shortens the timerequired. In addition, the method overcomes
the weak points of complicated and time-consuming pretreatment, and is expected tomaximize the advantages of GC/MS, such as
high separation, high qualitative ability, and afull database.
As a result of quantitative comparison (areavalue ratio) of each sample, differences bysample could be clearly determined. The dataobtained from the highly accurate analyticalmethodcancontributetohighly

reliableanalytical results, which can be utilized in the
future development of beer beverages andother products.
※To evaluate the reproducibility of the device, no correction
is made with internal markers, etc.