Sensitive Determination of 17 Organotin Compounds in Beverages Using Agilent 7890A / 7000B GC-MS/MS System

Agilent Technologies Wang Wenwen Cao Zhe

Background
Organotin compounds (OTCs) have been widely used as polymerized preservatives, insecticides and pesticides. The highest amount of OTCs found among coating materials in boats to prevent biological attachment in the ocean can reach to over 20% of the total which addressed great concerns. OTCs are identified as endocrine disruptive chemicals (EDCs) which may cause negative impact on human health via food chain. OTCs were found in ocean water, sediments, textiles and human urines however studies on OTCs in food were relatively less. There were studies on analysis of OTCs by GCMS (single Quadrupole) and ICPMS or LC-ICPMS, yet conventional GCMS method is with limited separation and sensitivity capacity, which can normally detect less than 10 OTCs in one run. So far, speciation studies of OTCs by GC triple Quadrupole mass spectrometer (GCMS/MS) technique are seldom seen.

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Background
Agilent 7000B GCMS/MS was applied in this research work to establish a precise, rapid and reliable method to study 17 OTCs in beverages. 11 kinds of beverages samples were tested to check preliminary contamination status of OTCs in the market which could be polluted by Tin containers.

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Experiment
Samples, Reagents Standards, Derivatization method

Sample Preparation
Chromatographic Parameters Mass Spec Parameters

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Samples, Reagents
The beverages were obtained from local grocery stores

Acetate buffer: 82g/L sodium acetate in water, adjusted to pH4.5 with acetic acid
Derivatization reagent :Dissolve 2g NaBEt4(Sigma-Aldrich ) in 10mL ethanol. This solution should be freshly prepared Ethanol, Hexane, Methanol chromatography grade purity

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Standards , Derivatization method

Standards: The standards were dissolved in methanol at 1000 ppm (1mg/mL) concentration and were further diluted, depending on the derivatization method used. Derivatization method: To 1 mL standard solution , 1 mL acetate buffer and 50 L derivatization reagent are added. The solution is shaken and allowed to react for 30min. After addition of 5mL water, the derivated compounds are extracted in 1 mL hexane. The mixture is vortexed for 10 s and the two phases are allowed to separate. The clear upper layer (apolar hexane phase) is transferred to an autosampler vial for analysis.

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Sample Preparation
Weigh 10 mL beverage sample into 22mL bottle (P.N 5183-4323), add 5.0 mL Methanol ultrasonic extraction 10 min.

To this solution, 2 mL acetate buffer and 200 L derivatization reagent are added. The sample is shaken and allowed to react for 30 min.

The derivated compounds are extracted in 1 mL hexane. The mixture is vortexed for 30 s and the two phases are allowed to separate. The clear upper layer (apolar hexane phase) is transferred and filtered to an autosampler vial for analysis.

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Chromatographic Parameters
GC system: Agilent 7890A Column: Agilent HP-5 MS UI capillary column (30 m0.25 mm0.25 m)P.N 19091S-433UI; Column temperature: 50 hold 1.5 min , at 10 /min to 300 , hold 1 min; Carrier gas: Helium; Flow rate: 1.1mL/min; Injection port temperature: 280 ; Injection volume: 2 L; Injection mode: Splitless, purge on after 1 min

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Mass Spec Parameters

Mass system : 7000 MS/MS;

Ion source: EI; Ion source polarity: Positive ion; Ionization voltage: 70 eV; Ion source temperature: 230; Interface temperature: 280; Collision GasNitrogen 1.5 mL/min; Quenching Gas Helium 2.25 mL/min;

Solvent delay: 2.0 min;

Optimized MRM Parameters:(See Table1)

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Table 1. Optimized MRM Parameters

No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Compound Trimethyltin Dimethyltin Monomethyltin Monobutyltin Tripropyltin Tetrapropyltin Dibutyltin Monophenyltin Monoheptyltin Tributyltin Monooctyltin Tetrabutyltin Diphenyltin Diheptyltin Dioctyltin Tricyclhexyltin Triphenyltin RT 2.72 3.86 5.34 9.45 10.38 11.35 11.66 12.92 13.29 13.54 14.49 15.16 17.78 18.00 19.83 21.78 21.80
Quantifier MRM transition (m/z) 165-135 179-151 165-137 179-151 193-151 207-165 179-151 255-199 179-151 207-151 179-151 235-179 303-275 249-151 263-151 351-197 233-151 Qualifier MRM transition(m/z) 163-133 151-135 193-165 179-123 193-123 207-123 263-207 255-277 179-123 207-123 179-123 291-179 303-197 249-123 263-123 349-195 233-123

Collision/eV 15;15 5; 10 5; 5 5; 10 5; 10 5; 10 5; 5 15; 5 5; 10 5; 15 5; 10 5; 10 5; 15 5; 15 5; 15 20; 20 5; 15

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Results and Discussion

Chromatographic separation results Calibration Curve, Linear Fit

Recovery Results
Analysis results of 11 Kinds of Beverages

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Chromatographic separation results

7000 Scan TIC

GC separation was finished in 22 min with 15 of 17 compounds ideally separated. Although the last two OTCs could not be separated as expected, extraction of transition ions and precise quantitation were accomplished by unique feature of GC-MS/MS given that appropriate ions were selected, extracted and monitored, at meantime, optimized GC and MS parameters were applied.

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Calibration Curve, Linear Fit

Calibration curves were prepared at 1.0-200 g/L(1.05.010.020.0 50.0100.0200.0 g/L 7 points)
No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Compound Trimethyltin Dimethyltin Monomethyltin Monobutyltin Tripropyltin Tetrapropyltin Dibutyltin Monophenyltin Monoheptyltin Tributyltin Monooctyltin Tetrabutyltin Diphenyltin Diheptyltin Dioctyltin Tricyclhexyltin Triphenyltin Linear range (g/L) 1-200 1-200 1-100 1-200 1-200 1-200 1-200 1-200 1-200 1-200 1-200 1-200 1-200 1-200 1-200 1-200 1-200 R2 0.999 0.995 0.999 0.999 0.996 0.996 0.999 0.998 0.999 0.999 0.997 0.998 0.998 0.999 0.999 0.998 0.999

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17 Organotin Compounds 1.0ppb MRM

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17 Organotin Compounds 5.0ppb MRM

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Monobutyltin 0.999

Dibutyltin 0.999

(g/ml)

(g/ml)

Tributyltin 0.999

Tetrabutyltin 0.998

(g/ml) (g/ml)

Organotin compounds Standard Calibration Curve Results (1)

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Monomethyltin 0.999

Monophenyltin 0.998

(g/ml)

(g/ml)

Trimethyltin 0.999

Tetrapropyltin 0.996

(g/ml) (g/ml)

Organotin Compounds Standard Calibration Curve Results (2)

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Recovery Results
Beverage samples were spiked with 10 g/L and 50 g/L mixed standard thus precision and recovery study of the method were conducted, majority of RSDs were within 10% and recoveries within 70-120%.
NO.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Compound Name Dibutyltin Diheptyltin Dimethyltin Dioctyltin Diphenyltin Monobutyltin Monoheptyltin Monomethyltin Monooctyltin Monophenyltin Tetrabutyltin Tetrapropyltin Tributyltin Tricyclhexyltin Trimethyltin Triphenyltin Tripropyltin 10g/L(n=3) AVE RSD 108.9 3.2 76.0 6.5 136.3 5.1 67.5 6.3 102.1 5.6 156.2 5.8 116.2 9.4 113.0 7.0 129.4 3.8 115.4 4.6 63.4 8.2 71.0 3.5 117.9 7.8 107.4 6.1 87.3 6.7 82.2 7.5 104.9 4.7 50g/L(n=3) AVE RSD 114.9 2.4 83.7 1.8 136.8 2.4 75.5 0.8 96.7 2.3 135.7 2.8 114.4 9.0 163.4 9.2 112.8 8.1 118.0 1.7 77.9 11.3 87.7 6.1 112.1 8.9 99.8 2.2 119.5 10.6 82.4 3.4 114.3 2.4

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Analysis results of 11 Kinds of Beverages

11 kinds of Beverages were then tested with above established method for the 17 OTCs among which both Dimethyltin and Monobutyltin were detected in 2 brands at sub ppb level.

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Analysis results of Sample 1

Dimethyltin

Monobutyltin

OT Dimethyltin Monobutyltin

RTmin 3.86 9.46

Concentrationg/L 0.27 0.37

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Analysis results of Sample 2

Dimethyltin

Monobutyltin

OT Dimethyltin Monobutyltin

RTmin 3.86 9.46

Concentrationg/L 0.27 0.39

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Summary
Agilent 7000 GCMS/MS technique with MRM mode has the advantage of eliminating background interferences, highly sensitive and selective, so that it turned out to be suitable and effective in analyzing 17 OTCs in beverages.

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Reference
Agilent application note :
Improving the Analysis of Organotin Compounds Using Retention Time Locked Methods and Retention Time Databases 5988-9256EN

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Thank you !!!

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