LaviStomaⓇ - Large
Large volume injection system with a liner in the Shape of a Stomach
StomachApplication of derivatiztion in stomach shapedliner

Y.K. KWON and R. SASANO

Saika Technological Institute Foundation, Wakayama-city, JAPAN[Introduction] Derivatization analysis is usually performed in gas chromatography. In this case, derivatization
proceeds in sample preparation step. But many of derivatization reagent are harmful for human body, and also,
derivatization process contains laborious steps. In this research, we have studied the possibility of completing
derivatization reaction in the stomach shaped liner of LaviStoma injection system, where injected sample is kept
as liquid.
[Method] We have studied these samples of 1ppm Pentachlorophenol(PCP), Bisphenol A, 2,4-Dichlorophenol(2,4-D)
and 10ppm fatty acids in 20% Acetone/Hexane. Derivatization reagent used here is 1% BSTFA(N,Obis(trimethylsilyl)trifluoroacetamide) in 20% Acetone/Hexane. Operating conditions of GC/MS and this injection
system are shown in Table 1. Using sandwich mode of Auto-injector, 20ul sample and 5ul derivatization reagent
are sucked in from each vial consequently, and then injected into the liner. The injector temperature is set justbelow the boiling point of the solvent to avoid its bumping. Sample is derivatized and concentrated while solvent isvaporized in split mode. Next, injector temperature is raised, and derivatized sample is introduced into column in
splitless mode. In this way, derivatization analysis is performed in GC/MS.
Temperature
Injector TemperatureHigh

1st stage

2nd stage

InjectingConcentrating

3rd stage

Transferring

4th stage

Eliminating

Solvent Boiling PointColumn Oven Temperature
LowSplit
1st stage
Injecting

2nd stage
Concentrating

TimeSplitlessSplit

3rd stage
Transferring

4th stage
Eliminating and Separating

Derivatization
reagent

SampleDerivatives

Table 1. Operating conditions of GC/MS and injector
Instrument

MainColumn

MSPreColumn

ColumnGC method

MS method

Fig. 1. Installation of large volume injection system: GCMS-QP5050A(SHIMADZU): AOC-201(SHIMADZU) with Sandwich method: Injector : LaviStoma(EMINET) with middle size liner(0.5ml): Pre-column : Deactivated silica capillary tube 0.53mm✕0.3m
Main-column : DB-5MS 0.25mm✕30m, 0.25µm: Injection Temp. : 69℃(0.5min)-100℃/min-260℃(18min)
Solvent Purge Time : 5sec
: GC Temp. : 60℃(5min)-15℃/-300℃(2min): SCAN Mode, 9-22min, 60-500m/z
Insertof Syringe

30.0e6TIC

Cl

25.0e6

1.0E+05

Fig. 2. Scheme of injection method using LaviStoma injection system

Peak AreaCl

10.0e6

1.0E+04

5.0e6

1.0E+031230.0e613141516

1.0E+020.0001TICCl

25.0e6

ClOH + BSTFA

Cl

20.0e6

ClCl

B

0.001

0.01

0.1

1Concentration (ng/µl)

TMS

ClFig. 4. Calibration curve of PCP-TMS
Cl15.0e6ClClCl

10.0e6

Table 2. Peak area of PCP-TMS of 7 consecutive analysis, theiraverage area and relative standard deviations(R.S.D.)

5.0e6

1213

1415

Compound16Fig. 3. Chromatogram of PCP and PCP-TMS. PCP-TMS is obtained asa result of derivatization in the stomach shaped liner of LaviStoma
injector. BSTFA is used as derivatization reagent.

TMS

23

4

56Ave.

7RSDSample : 0.01 ng/µl PCP 20µl

Derivatization reagent : 1% BSTFA 5µlOO

R‐COOH + TMS-Diaazomethane

TMS

R‐COO‐TMS
TMS

60000000

2,4-D

50000000

Bisphenol A

20000000

OCH2COOH+ BSTFA

Cl

Palmitic acid TMS

18000000

Cl40000000

OCl

16000000

TMS

Linoleic acid TMS

14000000ClIn tensity

A b s o lu te In te n s ity

1

PCP-TMS 11,206 10,993 11,146 11,285 11,393 11,038 10,910 11,139 1.53

OH + BSTFA

HO

70000000

30000000
20000000

Oleic acid TMS

12000000
10000000

Stearic acid TMS

8000000
6000000
4000000

10000000

2000000
00910

11

1213

14

15

16Ret.Time

17

Injectionofderivatization
reagent

R2 = 0.9985

OHCl

15.0e6

1.0E+06A

ClCl

20.0e6

Injectionof Sample

18192021221717.2

17.4

17.6

17.8

18

18.2

Retention time (min)Fig. 5. Chromatogram of TMS-derivatized Bisphenol A and 2,4-Dchlorophenol (left) and fatty acids (right). BSTFA is used as theirderivatization reagent.

18.4[Results] Figure 3 is a chromatogramof PCP (A) and PCP-TMS (B). Inchromatogram B, BSTFA is used as
derivatization
reagent
andderivatization proceeds in the liner ofLaviStoma injection system. Only
PCP-TMS peak, and no PCP peak, isfound in chromatogram B. Thisshows that PCP is fully derivatized
into PCP-TMS. Good linearity ofPCP-TMS between its concentrationand peak area is obtained as shownin figure 4. Also good repeatability ofthe peak area of PCP-TMS isobtained as shown in Table 2.
Moreover, Bisphenol A, 2,4-D andfatty acids are completely derivatized
as shown in figure 5. In this case,
BSTFA is used as derivatization
reagent.
[Conclusion] As mentioned above, we
can obtain results of derivatization
analysis quite easily and we need nolaborious preparation process ofderivatization before sample injection.
In conclusion, derivatization analysis
can be performed quite easily, simply
and quickly by using LaviStoma
injection system.