Význam tématu
V oblasti analytické chemie je přesné stanovení detekčního limitu přístroje klíčové pro posouzení citlivosti metod a zajištění spolehlivosti výsledků. Tradiční výpočet signál-šum (S/N) často nedostatečně zohledňuje variabilitu měření a může podléhat subjektivním zásahům při výběru šumových oblastí.
Cíle a přehled studie
Tato studie se zaměřuje na praktické stanovení a validaci instrumentálního detekčního limitu (IDL) na příkladu Thermo Scientific ISQ LT Single Quadrupole GC-MS. Hlavní cíle zahrnují porovnání tří samostatných přístrojových sestav z různých laboratoří a ověření reprodukovatelnosti metody pomocí standardu octafluoronaphthalenu (OFN).
Použitá metodika a instrumentace
IDL byl stanoven podle vzorce IDL = t × Amount × %RSD, kde t (2,998) je Studentův koeficient pro n=8, Amount představuje množství analyzovaného OFN (10 fg on-column) a %RSD relativní směrodatná odchylka z osmi opakovaných injekcí. Experimentální nastavení zahrnovalo
- GC-MS: Thermo Scientific ISQ LT Single Quadrupole
- Autosamplery: AS 1310 Liquid Autosampler a TriPlus RSH
- Kolona TraceGOLD SQC, 15 m × 0,25 mm × 0,25 µm
- Injekce 1 µL, splitless; helia 1,2 mL/min; program 40 °C (1 min) → 150 °C (30 °C/min) → 250 °C (100 °C/min)
- MS: EI 70 eV; DynaMax XR detektor; SIM m/z 272; transfer line 250 °C; ionizátor 200 °C
Hlavní výsledky a diskuse
Na všech třech systémech provedených v Singapuru, Itálii a UK byla naměřena opakovatelnost špičkových ploch s %RSD pod 4 % a vypočtené IDL se pohybovalo mezi 1,0 fg až 1,2 fg. Výsledky potvrzují vysokou citlivost a stabilitu IDL bez ohledu na stáří přístroje či typ autosampleru. Kombinace S-tvaru iontového vedení a DynaMax XR detektoru minimalizuje chemický a neutrální šum.
Přínosy a praktické využití metody
Statistický výpočet IDL poskytuje objektivní hodnocení citlivosti GC-MS metod, usnadňuje srovnání mezi přístroji a podporuje splnění regulačních požadavků. Metoda je vhodná pro QA/QC laboratoře, výzkumné studie i průmyslové aplikace stopové analýzy.
Budoucí trendy a možnosti využití
Dynamický rozvoj iontových vedení a detekčních systémů a automatizace výpočtů přinese další snížení detekčních limitů. Plánováno je rozšíření metody na komplexní vzorkové matrice a integrace pokročilé analýzy dat pro zvýšení efektivity.
Závěr
Thermo Scientific ISQ LT Single Quadrupole GC-MS vykazuje vynikající detekční limity pod 1,5 fg on-column s vysokou reprodukovatelností mezi různými systémy. Statistický přístup k výpočtu IDL zajišťuje spolehlivou charakterizaci citlivosti přístroje.
Reference
- US Code of Federal Regulations: definice MDL, 49 FR 43430 (Oct. 26, 1984); 50 FR 694, 696 (Jan. 4, 1985), novelizováno na 51 FR 23703 (June 30, 1986).
- IUPAC Analytical Compendium: definice detekčního limitu, sekce 2.2.1.
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Practical Determination and Validation
of Instrument Detection Limit of
Thermo Scientific™ ISQ™ LT Single
Quadrupole GC-MS
Richard Law, Senior Application Scientist, Thermo Fisher Scientific, Runcorn UK
Tommaso Albertini, Marketing Manager, Thermo Fisher Scientific, Rodano Italy
RT: 11.60 - 12.00
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RT: 11.93
SN: 3671RMS
NL:
1.85E6
m/z=
283.5-
284.5 MS
ICIS
HCB_Sig_
To_Noise
×250
Figure 1.
Noise range 11.7-11.8
RT: 11.60 - 12.00
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NL:
1.85E6
m/z=
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ICIS
HCB_Sig_
To_Noise
×250
Noise range 11.75 – 11.85
Figure 2.
Chromatograms show the effect of noise range selection on S/N value calculated.
Introduction
Signal to Noise (S/N) has been used
for decades to convey the detection
limit of an instrument. Technological
innovation has driven instruments to
exhibit lower noise and greater
sensitivity; whilst advancements in
gas separation technology mean
lower bleed columns and sharper
peaks, again, leading to significant
improvements in S/N values
obtained. Moreover, S/N calculations
are also typically performed on a
single injection, not demonstrating
the repeatability of the instrument in
question. S/N measurements usually
always rely on some form of operator
involvement, such as selecting the
start and end of the peak being
measured and more importantly
choosing the noise range. Figures 1
and 2 show the differences in S/N
value for the m/z 284 fragment ion
of hexachlorobenzene when the
selected noise range differs by only
0.05 minutes. On many GC triple
quadrupole instruments the added
selectivity leads to much lower
baseline noise, rendering the use
of S/N as an accurate measure of
instrument detection limit inadequate.
Therefore, a more accurate or
statistically significant method of
calculating the detection limit of an
instrument is required.
IDL = t * Amount * %RSD
Where,
t = student t-value for one-tailed distribution:
for n = 8 injections; t = 2.998
Amount = amount of analyte (on-column)
%RSD = relative standard deviation of the response
During this study, the consistency in IDL for octafluron-
apthalene (OFN), a commonly used reference compound
in GC-MS, was determined on three separate Thermo
Scientific ISQ LT Single Quadrupole GC-MS systems
using the method described above.
2
Instrument Detection Limit (IDL)
One way of determining the instrumental Limit of
Detection (LOD) is by using the standard deviation of
the response of an analyte of choice at a concentration
close to the estimated detection limit. This approach gives
much more reliable LOD values as it takes into account
not only the signal intensity, but also the consistency of
the response. Method Detection Limits (MDL) are firmly
established with many regulatory bodies and can be
defined in many ways. According to the most recent
U.S. Code of Federal Regulations1, MDL is defined as:
“the minimum concentration of a substance that can be
measured and reported with 99% confidence that the
analyte concentration is greater than zero and is deter-
mined from analysis of a sample in a given matrix
containing the analyte.” This methodology can be
seamlessly transferred when calculating Instrument
Detection Limits (IDL).
Unlike MDL, the IDL uses solvent based standards
containing the test chemical at concentrations that give
a consistent response over several repeat injection
(ex: %RSD <5). Therefore, the IDL is a statistically
rigorous method that uses the precision of a measurement
at low analyte levels and accurately reflects the true
detection limit of an instrument, ultimately defining how
sensitive an analytical system is. Simply, the IDL can be
determined by using the following equation2:
GC Conditions
Instrument
Thermo Scientific™ TRACE™ 1310 ISQ™ LT Single Quadrupole GC-MS
Column
Thermo Scientific™ TraceGOLD™ SQC GC Column (p/n: 26070-1300)
Column dimensions
15 m × 0.25 mm × 0.25 µm
Injection volume
1 µL
Inlet temperature
220 °C
Injection Liner
Thermo Scientific™ LinerGOLD™ Splitless Liner single taper with wool, 4 mm ID, 78.5 mm length (p/n:
453A0924)
Injection mode
Splitless
Split flow rate/Splitless time
50 mL/min at 0.8 minutes
Septum purge flow
5 mL/min (constant)
Carrier mode
Constant flow
Carrier gas
Helium
Carrier flow
1.2 mL/min
Temp. gradient
40 °C (hold for 1.0 min); ramp 30 °C/min to 150 °C; ramp 100 °C/min to 250 °C; (hold for 5 min)
Total run time
10.67 min
MS Conditions
Ionization mode
EI, 70eV
Emission Current
50 µA
MS transfer line temperature
250 °C
Ion source temperature
200 °C
Acquisition mode
Selective Ion Monitoring (SIM) m/z 272
Software (CDS)
Dionex™ Chromeleon™ 7.2 Chromatography Data System
Table 1.
Experimental
Thermo Scientific ISQ LT Single Quadrupole GC-MS
systems were used in all of the experiments, with AS 1310
Liquid Autosampler (two experiments) and a TriPlus RSH™
Autosampler (one experiment). Choosing different robotic
arms on ISQs used for this test adds to the rigorosity of
the experimental setup and the reduction of any potential
autosampler bias. See Table 1 for detailed conditions.
All experiments were performed using a 1 µL injection
solution containing 10 fg/µL octafluoronaphthalene
(OFN) in iso-octane. The IDL was calculated from n=8
sequential replicate injections.
3
Results and Discussion
Three different ISQ LT Single Quadrupole GC-MS
systems were evaluated using eight sequential replicate
injections of a 10 fg/µL octafluoronaphthalene standard in
iso-octane. The results from each instrument, along with
calculated relative standard deviations from which the
IDL is derived, are summarized in Table 2.
The instruments used in the experiment are based in three
different laboratories around the world and have been
deployed in environments running routine applications,
in instrument engineering settings, and as training
instruments for a number of years (Figure 3). Three
separate operators were responsible for setting up the
systems, using three different standard preparations and
utilizing two different autosampler types. Even with these
variations, all three instruments demonstrated peak area
repeatability with %RSD < 4% and displayed outstand-
ingly consistent IDL values, regardless of their age and
typical usage. The combination of the ISQs off-axis ion
source and unique s-shaped ion-guide creates a curved
path allowing chemical and neutral noise to be kept to a
minimum, enabling lower detection limits, whilst the
Thermo Scientific DynaMax XR detection system provides
the sensitivity to detect and amplify the low number of
transmitted ions needed at this challenging analyte
concentration.
Injection
Peak area (counts per second)
Instrument 1
Singapore
AI/AS 1310 ISQ LT
Instrument 2
Italy
AI/AS 1310 ISQ LT
Instrument 3
UK
TriPlus RSH ISQ LT
1
415.02
315.48
249.12
2
417.78
326.94
244.68
3
402.36
304.92
245.52
4
393.48
289.20
266.34
5
412.26
301.92
253.32
6
403.26
308.22
260.88
7
372.12
307.98
266.16
8
388.68
319.80
257.28
Average
400.62
309.31
255.41
SD
15.37
11.59
8.66
%RSD
3.8%
3.7%
3.4%
IDL (fg)
1.2
1.1
1.0
Table 2.
TN10494-EN 0816S
Conclusion
The Thermo Scientific ISQ LT Single Quadrupole
GC-MS system exhibits outstandingly low instrument
detection limits ensuring confidence in your analysis.
The use of a statistical method to determine the IDL
demonstrates the consistent and sensitive detection that
can be achieved at this challenging analyte concentration.
The instrument-to-instrument detection limits using a
10 fg on-column amount have been shown to be highly
reproducible and consistent in a variety of settings,
regardless of age and use.
The exceptionally sensitive Thermo Scientific DynaMax
XR detection system, complemented by the innovative
S-Shaped Ion Guide, effectively reduces noise whilst
maintaining maximum sensitivity leading to IDLs
significantly below 1.5 fg on column.
References
1. U.S. Code of Federal Regulations, 49 FR 43430, (Oct.
26, 1984); 50 FR 694, 696 (Jan. 4, 1985), as amended
at 51 FR 23703 (June 30, 1986), http://www.ecfr.gov/
cgi-bin/text-idx?SID=efe93db42854f88dffcf66ba8de73
7e6&mc=true&node=ap40.23.136_17.b&rgn=div9
2. International Union of Pure and Applied Chemists
(IUPAC), http://www.iupac.org/publications/analytical_
compendium/Cha18sec437.pdf
Figure 3.
UK
Italy
Singapore
Injections
Peak Area (OFN)
IDL 1.0fg
Injections
Peak Area (OFN)
IDL 1.1fg
IDL 1.2fg
Injections
Peak Area (OFN)
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