SPL-M100 Application
SPL-M100
ApplicationNoteNoteNo. AS240508EN

Metabolome analysis of chicken liverby online solid-phase analytical derivatization GC-MS system
Introduction

for SPE-GC system

Solid-phase derivatization (SPD) is a techniqueofderivatization

without

theSample

time-consuming

centrifugal concentration and lyophylization, bySpiral insert

retaining the target compounds on a solid phase

Flash-SPEand dewatering it by passing a organic solvent
throughitthenSPL-M100
SPL-M100

infiltrating

theLiver(chicken)

derivatization

reagent and performing the reaction on the solidphase. The example of pretreatment method andanalytical condition for metabolome analysis ofInformation
Information

chicken liver are shown below.
Online SPE-GC-MS system

SPD pretreatment workflow
Sample 1 g

Online SPE-GC: SPL-M100

Vials

Water 1 mL
Transfer 200 µL

Flash-SPE ACXs

ACN 800 µL

Loading [Retaintion] Extract 50 µL
Washing ACN-Water (1/1)

Shaking (37 ℃, 30 min)Dewater ACN

Centrifuge (14000 rpm, 3 min)Infiltration 0.5% Methyoxyamine/Pyridine

Supernatant 200 µL

Derirvatization (Methoximation), 3 min

ACN-Water (4/1) 800 µL

Infiltration MSTFA/Hexane (1/1)
Derivatization (TMS), 1 min

Analytical solution

Elution Hexane

pH adjustment (8-9)
GC-MS

Analytical condition
SPE-GC interface
SPE cartridge
PTV injection port

AutomationConditioning
ACN-Water (1/1)

SPL-M100 (AiSTI SCIENCE)
Flash-SPE
LVI-S250 (AiSTI SCIENCE)

Insert typeSpiral insert

Temp.

220℃(0.5 min)-50℃/min-290℃(23 min)Gas chromatograph
Inlet mode

Split 1:50

Flow mode

Constant flow, 1.0 mL/min

Pre-column

0.25 mm i.d. x 0.5 m

ColumnVf-5ms, 0.25 mm i.d. x 30 m, df=0.25 μm

Oven Temp.

100℃(2 min)-10℃/min-320℃(2min)

Transfer line Temp.

290℃

Mass spectrometer
Acquisition mode

Scan (m/z 70-600)

Data acquisition

3.0-26 min

AiSTI SCIENCEAiSTI SCIENCE
Product

Online SPE-GC

SPL-M100
Solid-phasecartridge

ProductFlash-SPE
GC large volume
injection port

LVI-S250

AiSTI SCIENCE Co., Ltd.
TEL: +81-73-475-0033
E-mail: as@aisti.co.jp
Tel：+81-73-475-0033
www.aisti.co.jp

AiSTI SCIENCE CO.,Ltd.

E-mail：as@aisti.co.jp
HP：www.aisti.co.jp

No. AS240508EN

AiSTI Application NoteResults

ExpandedFigure: Total ion current chromatogram
Table: Result of repeatability test (n=5, RSD%)
No.
12345678910111213141516171819202122232425262728

化合物名Metabolites
Lactic acid_2TMS
Alanine_2TMS
Valine_2TMS
Phosphate_3TMS
Isoleucine_2TMS
Proline_2TMS
Glycine_3TMS
Serine_3TMS
Threonine_3TMS
beta-Alanine_3TMS
Aspartic acid_3TMS
Methionine_2TMS
Hydroxyproline_3TMS
Pyroglutamic acid_2TMS
Glutamic acid_3TMS
Phenylalanine_2TMS
Asparagine_3TMS
Glutamine-TMS
L-Glutamine-3TMS
Lysine_4TMS
Tyrosine_3TMS
Gluconic acid_6TMS
Palmitic acid-TMS
Uric acid-4TMS
Linoleic acid-TMS
Oleic acid-TMS
Stearic acid-TMS
Arachidonic acid-TMS

鶏肝-1
1
9,262,000
18,330,000
11,330,000
13,080,000
7,530,000
13,220,000
11,330,000
4,217,000
2,152,000
1,152,000
8,020,000
2,438,000
37,2204,924,000
8,770,000
2,531,000
173,700
529,400
3,871,000
523,000
6,123,000
198,100
843,200
3,099,000
95,410341,900
385,000
31,050鶏肝-2
28,665,000
17,960,000
11,100,000
14,290,000
7,478,000
13,140,000
11,300,000
4,070,000
2,096,000
1,125,000
7,178,000
2,365,000
34,9804,513,000
7,753,000
2,438,000
163,500
583,500
3,613,000
481,100
5,808,000
157,500
724,400
2,251,000
76,620280,500
325,700
25,380鶏肝-3
39,610,000
18,290,000
11,340,000
15,760,000
7,716,000
13,420,000
11,770,000
4,218,000
2,153,000
1,193,000
7,640,000
2,490,000
39,9104,615,000
8,353,000
2,570,000
158,000
634,500
3,810,000
535,400
6,279,000
190,100
808,800
2,557,000
81,480310,400
348,700
26,610鶏肝-4
48,632,000
18,100,000
11,280,000
15,190,000
7,693,000
13,360,000
11,600,000
4,167,000
2,152,000
1,174,000
7,525,000
2,486,000
36,0904,781,000
8,250,000
2,541,000
170,000
518,900
3,810,000
523,600
6,303,000
171,900
788,200
2,424,000
76,570286,900
349,900
24,410鶏肝-5
59,583,000
18,150,000
11,340,000
15,900,000
7,683,000
13,450,000
11,440,000
4,180,000
2,145,000
1,161,000
7,633,000
2,490,000
36,8104,571,000
8,426,000
2,575,000
169,100
643,500
3,906,000
524,500
6,438,000
195,200
825,600
2,676,000
84,040323,400
360,000
26,570Ave.
RSD, %
9,150,400
5.2
18,166,000
0.8
11,278,000
0.9
14,844,000
7.9
7,620,000
1.4
13,318,000
1.011,488,000
1.7
4,170,400
1.4
2,139,600
1.1
1,161,000
2.2
7,599,200
4.0
2,453,800
2.2
37,0025.0
4,680,800
3.6
8,310,400
4.4
2,531,000
2.2
166,860
3.7
581,9609.9
3,802,000
3.0
517,520
4.1
6,190,200
3.9
182,560
9.5
798,0405.8
2,601,400
12.3
82,824
9.3
308,6208.2
353,860
6.1
26,8049.5