For metabolome analysis

Online SPE-GCsystem

SPL-M100■ Dimensions (unit: mm)
Main unit

410520Liquid delivery part390340■ SPL-M100 specifications
Dimensions

Solid phasederivatization
methodMain unit

W 340 mm, D 560 mm, H 520 mm (from the installation surface)

Liquid delivery part

W 390 mm, D 570 mm, H 410 mm

Power supply and power consumption 100 V (500 VA)
PC specs for software

Windows XP or later

Number of specimens processed Up to 50 specimens
Liquid delivery

Syringe method

Gas usedN2 gas or inert gas

Installation environment

Temperature: 18-28°C
Humidity: 40-70% RH, but no condensation.
Other: It is desirable to have an environment with minimal interference elements
such as dust, vibrations, ambient noise, corrosive gases, etc.
Separately, the LVI-S250 Large Volume Injector for GC (sold separately) is required.
(If you have LVI-S200, you can upgrade to LVI-S250.)

Product specifications, appearances, configurations, etc. may be changed without notice for improvement.
Company names and product names in this catalog are registered trademarks or trademarks of the respective companies.

Please contact us or our distributors if you have any questions about our products.
XX-1005, December 24, 2021 edition

AiSTI SCIENCE Co., Ltd.
18-3 Arimoto, Wakayama City, Wakayama Prefecture, 640-8390
Tel: (073) 475-0033 Fax: 073-497-5011
Adobansu 610, 1-1-31 Hamasaki, Asaka City,
Saitama Prefecture 351-0033
Tel: (048) 424-8384 Fax: 073-497-5011(Headquarters: Wakayama)

www.aisti.co.jp

Online SPE-GC/MS system forsolid phase derivatization method
Derivatization revolution
What used to take 2 days now
just takes 10 minutes!

Solid phasederivatization■ It is expected that the “solid phase derivatization method”
can be used in a variety of fields.

Medical and pharmaceutical

Dehydration
1 minute

Derivatization
2 minutes

SPL-M100

STEP ❶STEP ❷STEP ❸STEP ❹STEP ❺

Sample loading& retention

Washing

Dehydration/ 1 minute

Derivatization/ 2 minutes

ElutionAmino acids and organic
acids are retained in thesolid phase by ion
exchange interactions

Removing the remaining
non-ionic compounds inthe solid phase withwater-ACN

Removing the remaining
water in the solid phasewith ACN

Derivatization of the target
component in the solidphase by impregnating
the solid phase with aderivatization reagent

SampleWater-ACN

ACN-SO3⊖-N⊕(CH3)3Amino group⊕H3N-R

Carboxyl group⊖OOC-R

Flash-SPE cartridges enablingfull-volume injection
A solid phase cartridge developed
specifically for online SPE-GC. With a very
low filling volume of 2 to 5 mg, thecompact design of the Flash-SPE and the
LVI-S250 large volume injector enable

Hexanefull-volume injection of samples.

⊕H3N-R-SO3⊖⊕H3N-R-SO3⊖

TMS-HN-R

-SO3⊖

-N⊕

⊖OOC-R-N⊕⊖OOC-R-N⊕

TMS-OOC-R-N⊕

Flash-SPE

R-NH-TMS
R-COO-TMS(Solid phase mini cartridge)Non-ionic compounds

02 AiSTI SCIENCE SPL-

Solid-phase extraction equipment is mounted
on GC/MS to achieve fully-automated online processing.
Fully-automated through GC/MS analysis by just setting the sample.

-SO3⊖(CH3)3(CH3)3

H2O(CH3)3Grain harvesting■ Online SPE-GC system for metabolome analysis

Solid phase derivatization method Patent registration

MSTFA

Foods and drinks(CH3)3

Flash-SPE

AiSTI SCIENCE SPL-M10003

■ Online solid phase derivatization method

Hardware functions and features
Capable of direct large volume injection of samples via robotic arm.
Existing sample trays can be used as-is.
It can also be used for conventional analysis by replacing the existing autosampler.
Sensors are installed at various locations to prevent operational problems.

STEP ❶STEP ❷Sample loading& retention

WashingAmino acids and organic
acids are retained in thesolid phase by ion
exchange interactions

Removing the remaining
non-ionic compounds inthe solid phase withwater-ACN

SampleWater-ACN

STEP ❸STEP ❹STEP ❺Dehydration Derivatization/ 1 minute/ 2 minutes
Removing the remaining
water in the solid phasewith ACN

Derivatization of the target
component in the solid phase byimpregnating the solid phasewith a derivatization reagent

ACN

MSTFA

ElutionHexaneSyringe pump

Solid-phaseextraction
Flash-SPE■ Current status of pretreatment and derivatization inmetabolomics and suggestions for improvement
About pre-treatment operations
The current major challenge is that pretreatment
processes such as centrifugal concentration, freeze-drying,and derivatization are complicated, time-consuming, andrequire skilled techniques and experience.

GC inlet LVI-S250

GC/MS
Spiral (stomach-shaped) insert

Solid phase derivatization method forfaster speed and automation

Non-ionic compounds

H2O

About measurement of multiple samples
When multiple samples are derivatized in a batchprocess, the time from derivatization to measurement
differs for each sample.

Onlinefrom derivatization to measurement via SPE-GC■ Fully automated from pretreatment (automatedpretreatment time of 5-10 minutes) toGC/MS analysis
Dehydration

Extraction
Analysis

Derivatization

Full-volumeinjection

Simply place
the sample in a vialand set it in theSPL-M100

The time between
derivatization andinjection is always
constant■ Various nozzles enable multi-stage liquid flow
Smooth multi-stage liquid flow processing is performed by using dedicated nozzles for eachprocess (conditioning, sample aspiration loading, washing, nitrogen ventilation, and elution).

■ Working layout
designed forefficiency

Sample setting area

A working layout designed to fully
automate processing from solid phaseconditioning, purification, drying, andelution to GC/MS injection.
In addition, a syringe pump dedicated foreach solvent is installed for liquid delivery,
eliminating concern of mixing liquids.

Working area

Elution and injection area

SPE-GC/MS■Use of autosampler
By separating the base plate, the front inlet
can be used with an existing autosampler.

■ Sample setting area (sample collection)
UP TO

50Sample・Elution

04 AiSTI SCIENCE SPL-

■ Flash-SPE solid phase setting area
UP TO

100

Flash-SPE■ Inject into GC at the sametime as elution■ Solid phase disposal
receptacle

AiSTI SCIENCE SPL-M100

05

Performance evaluation of solid
phase derivatization method■ Simultaneous analysis of amino acids, organic acids,
and nucleobases● SCAN total ion chromatogram

Fig. SCAN total ion chromatogram of mixed standard aqueous solutions (concentration in vial:
0.02nmoL / μL) obtained by the online solid phase derivatization SPE-GC/MS system[Chromatogram] TIC: 5977585-024,25

113.41057621● SCAN total ion chromatogram
Fig. SCAN total ion chromatogram of mixed standard aqueous solutions (concentration in vial:
0.02nmoL / μL) obtained by the online solid phase derivatization SPE-GC/MS system[Chromatogram] TIC: 14491961-0

1015,1612817232028

9137261422918127192134

5689111222● ReproducibilityTable: Peak area values and their RSD (%, n=9) for mixed standard aqueous solutions obtained by
the online solid phase derivatization SPE-GC/MS system

06 AiSTI SCIENCE SPL-

■ Simultaneous analysis of saccharides● Linearity
Fig. Relationship between
concentration and area value● Effect of pH
Fig. Relationship between pH andrelative area value

AiSTI SCIENCE SPL-M10007

■ Mouse serum● Additive recovery test

1st: Extraction

2nd: Automated derivatizationand measurement● Sample: 50 μL (50 mg)

Table: Recovery rate (%) obtained by adding tomouse serum extract● Sample loading volumeFig. Relationship between sample loading
volume (10, 20, 40, 80 μL) and peak areavalues● Vial・Added water: 150 μL
・Added ACN: 800 μL● Fractionation: 40 μL● Shaking (37 ℃, 30 min）

● Flash-SPE AOS-3mg● Centrifugal separation(14,000 rpm, 5 mins)

Simply set thevial in thesample tray

● Fractionation:
Supernatant solution: 250 μL
・Added water: 250 μL● Overlay chromatograms ofquantitative ions・Conditioning
・Water: 100 μL
・Acetonitrile: 100 μL
・Washing water-ACN (1/4): 100 μL
・Dehydration ACN: 100 μL

Fig. Overlay chromatograms of quantitativeions of mouse serum obtained using thesystem (n=9)

・Impregnation Methoxyamine-Pyridine-MSTFA: 8 μL

● Solid phase derivatization

reaction: 2 mins

● Vial・Elute acetone-hexane (1/4): 40 μL● GC/MS: LVI-Split (1:50)● SCAN total ion chromatogram● Reproducibility

Table: SCAN total ion chromatogram for mouse serum obtained by the online solid phase
derivatization SPE-GC/MS system[Chromatogram] TIC: 4331844-0

Table: Peak area values and their RSD (%, n=9) for mouse serum obtained by the online solid phase
derivatization SPE-GC/MS system

3,4710111961220524,25272815808 AiSTI SCIENCE SPL-M100

1312

AiSTI SCIENCE SPL-M100

09

Software functions
and features

Flexible withconvenientfeatures● Intuitive and easy-to-use software

Large volume injector for GC with spiral(stomach-shaped) insert

LVI-S250
For Gas Chromatography

The ability to print sequences, logs,
and more helps in document management
and equipment management.

メニューバー メソッド・シーケンス作成や環境設定。

■ High-sensitivity analysis of low-concentration
samples due to greatly improved sensitivity■ Faster pretreatment by reducing sample volume
and omitting concentration operations■ Derivatization is possible in the insert■ Compatible with various injection methods,including large volume injection

4cm

動作状況をリアルタイムで表示。

Controller box

Large volume injection split mode

Flash-SPE
モニター

Inlet main partTarget substanceSpiral insert

Impurities

Volume: 0.6 mL*Actual size
Sample is held in the insert in aliquid state for flexible
concentration. Also, because it
can be introduced into thecolumn at a low temperature, it
can be used for heat-sensitive
substances.

スケジュール
スケジュールを直 接 入力（シーケンスファイルとして保存可能）。
処理したいサンプルにチェックを入れ、スタートをクリック。

● Method creation is possible■ List of preset registered methods○ Simultaneous analysis of amino acids,organic acids, nucleobases, and saccharides● Various log functions record
operating conditions■ Operation log & error log

Log files are automatically saved in one-monthincrements for easy management.

Also possible to create other original
methods. By registering frequently
used methods, anyone can easily workwith them.

Removing

Set the inlet
temperature lower
than the boiling pointof the elution solventand inject the eluate.

The elution solvent
is discharged whileevaporating toconcentrate thetarget component.

Increase the inlet
temperature andintroduce the target
component into thecolumn while splitting.

High-boiling
impurities areremoved by split.

Low filling volume for efficient analysis

For Online SPE-GC(Solid phase mini cartridge)
Piping■ Linear structure for smooth flow of sample and solution■ Fast ventilation drying (30 seconds)■ Simple structure optimized for automation■ Small solid phase filling volume of 2-5 mg■ Piping and needle can be connected from both top and bottom ends

Purpose of use andtype of solid phase

SPL-M100*Actual size
Frit

○ Analysis of fatty acids○ Analysis of saccharides

4th Stage

Split introduction

Various print-outs○ Analysis of organic acids○ Analysis of nucleobases

3rd Stage

Concentration

Solid phase cartridges for online SPE-GC○ Simultaneous analysis of amino acids
and nucleobases○ Analysis of amino acids

2nd Stage

Injection

Flash-SPE
Patent registration○ Simultaneous analysis of amino acids,organic acids, and nucleobases

1st StagePCPrinter■ Print functions

Sequence log

Error log

FritConnecting function

The ability to print sequences, logs, and more
helps in document management and equipment
management.

Solid phase (filler)
C-18, etc.

Straight structure forsmooth liquid flow anddrying. Efficient
pretreatment is realized.

Target10 AiSTI SCIENCE SPL-M100

Time of elution
and GC injection

Product nameFlash-SPE
ACXs

Flash-SPE
CXs

Flash-SPE
AXs

Amino acids●●

×Amines●●×

Nucleobases

●

●

×

Organic acids

●

×

●

Fatty acids

●

×

●

Saccharides

●

×

●

NeedleBoth ends are press-fitted.
Designed to enable firmconnection and flexibility.

*Quantity is 100 each

AiSTI SCIENCE SPL-M10011