1-(C12-C18 even numbered, C18 unsaturated)alkyl-1,4,5,6-tetrahydropyrimidin-2-aminium acetate and{[3-((C12-C18 even numbered, C18 unsaturated)alkylamino)propyl]amino}(imino)methanaminium acetate and[(3-{[ammonio(imino)methyl]amino}propyl)(C12-C18 even numbered, C18 unsaturated)alkylamino](imino)methanaminium diacetate

Adsorption / desorption

Administrative data

Endpoint:

adsorption / desorption: screening

Remarks:

adsorption

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-07-27 to 2012-12-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: HPLC estimation method.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: ISO 18749:2004(E)

Deviations:

yes

Remarks:

Bentonite was used as additional test system.

Principles of method if other than guideline:

Bentonite was used as additional test system.

GLP compliance:

yes

Type of method:

HPLC estimation method

Media:

other: activated sludge, bentonite, glass

Test material

Test material information

Constituent 1

Reference substance name:

C12-C16 (even numbered) alkyl-1,4,5,6-tetrahydropyrimidin-2-aminium acetate and {[3-(C12-C16 (even numbered)alkylamino)propyl]amino}(imino)methanaminium acetate and [(3-{[ammonio(imino)methyl]amino}propyl)-C12-C16 (even numbered)alkylamino](imino)methanaminium diacetate

EC Number:

939-650-3

Cas Number:

2770917-88-7

IUPAC Name:

C12-C16 (even numbered) alkyl-1,4,5,6-tetrahydropyrimidin-2-aminium acetate and {[3-(C12-C16 (even numbered)alkylamino)propyl]amino}(imino)methanaminium acetate and [(3-{[ammonio(imino)methyl]amino}propyl)-C12-C16 (even numbered)alkylamino](imino)methanaminium diacetate

Radiolabelling:

no

Study design

Test temperature:

SampleMeasured temperature[°C]Min21.2Max23.7Mean22.5SD ±0.8

HPLC method

Details on study design: HPLC method:

EQUIPMENT- Apparatus:Autosampler Acquity UPLC, WATERSBinary Solvent ManagerAcquity UPLC, WATERSColumn ManagerAcquity UPLC, WATERSDetector Mass selective detector, Xevo TQD MS, Acquity UPLC, WATERSSoftware MassLynxTM 4.1, SCN 791, WATERSAnalytical ColumnAcquity UPLC BEH C18 1.7 µm, 50 x 2.1 mm, batch 0199, serial number: 019931343257 84, WATERSReagents Acetonitrile, gradient grade, VWR HPLC water, gradient grade, VWR Trifluoroacetic acid (TFA), ≥ 99.9 %, ROTHExternal standardThe test item was used as external standard.CONDITIONS OF ANALYSISDetection Type:MRMIonisation ModeES+Capillary Voltage0.6 kVSource temperature150 °CCone gas flow (N2)80 L/hDesolvation temperature550 °CDesolvation gas flow (N2)1000 L/hCollision gas pressure (Ar) 3.11 * 10-3 mbarColumn Temperature30 °CMobile phaseA:HPLC-water with 0.1 % TFAB:Acetonitrile with 0.1 % TFATable 2:Gradient TableTime[min]A[%]B[%]0.050500.150503.020803.150504.05050The first 0.4 min of each run will be wasted to minimize possible matrix effects.Flow rate 0.6 mL/minRun time 4 minInjection volume10 µLMass spectrometric conditionsTable 3:Mass Spectrometric ConditionsAnalyte1-dodecyl tetrahydro pyrimidin-2(1H)-imine1-(3-(dodecyl amino)propyl) guanidine1-dodecyl- 1-(3-guanidino propyl) guanidinePrecursor ion [Da]268.2355285.2621327.2839Product ion, Quan [Da]100.0198100.0196310.3114Product ion, Sec [Da]57.827258.0168268.2979Dwell time [s]0.0240.0240.024Cone Voltage [V]725248Collision energy, Quan [eV]282218Collision energy, Sec [eV]343424REFERENCE SUBSTANCES- Identity: Basic Violet 4SynonymEthyl VioletDETERMINATION OF RETENTION TIMES- Quantity of test substance introduced in the respective incubation vessels: 10 mL- Quantity of reference substances: 5 mg/L (nominal)EVALUATIONDegree of adsorption:Concentrations below the limit of quantification (LOQM) have not been taken into account. Due to analytical reasons nominal test item concentration were used as t0 values.Distribution coefficientThe calculation of the distribution coefficient for the adsorption of 1-dodecyltetrahydropyrimidin-2(1H)-imine on glass surfaces is the only feasible, due to almost complete or lack of adsorption. The adsorbed concentration was related to the glass surface given in cm² and calculated.:

Batch equilibrium or other method

Analytical monitoring:

not required

Results and discussion

Adsorption coefficient

Type:

Kd

Value:

ca. 0.002 other: L/ cm²

Matrix:

glass surface

Remarks on result:

other: , no information on org. carbon% is available

Partition coefficients

Remarks on result:

other: other

Remarks:

The coefficient of determination (r2) of each of the calibration curves was ≥ 0.992.

Results: HPLC method

Details on results (HPLC method):

- Retention times of reference substances used for calibration: Calibration Data of the Reference Items for Soil and Sewage SludgeReference ItemReplicateExtinktion at 595 nmRRAt0 h24 h[%][%]10.6620.0152.397.720.6950.0172.497.6Concentration and Adsorption of 1-dodecyltetrahydropyrimidin-2(1H)-imineon Activated SludgeSampleReplicateSampling Interval0 h 3 h 6 h Calc. Conc.[µg/L]At[%]Calc. Conc.[µg/L] At[%]Calc. Conc.[µg/L] At[%]FT192010125 88< LOQM 1002929 9126 88< LOQM 100Mean92510126 88< LOQM 100Concentration and Adsorption of 1-dodecyltetrahydropyrimidin-2(1H)-imine on BentoniteSampleReplicateSampling Interval 0 h 3 h6 h Calc. Conc.[µg/L]At[%]Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L]At[%]FM1970 5< LOQM 100< LOQM100291411< LOQM 100< LOQM100Mean942 8< LOQM 100< LOQM100Concentration and Adsorption of 1-dodecyltetrahydropyrimidin-2(1H)-imine on Glass SurfaceSample Rep Sampling Interval0 h 3 h 24 h48 h 72 h Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%]FS 1821 20827196923244656 579 43 2850 17778245944259342 378 63 Mean836 19803226433752049 479 53Concentration and Adsorption of 1-(3-(dodecylamino)propyl)guanidine on Activated SludgeSampleReplicateSampling Interval0 h 3 h6 hCalc. Conc.[µg/L] At[%]Calc. Conc.[µg/L] At[%]Calc. Conc.[µg/L] At[%]FT18291913487< LOQM10029051113387< LOQM100Mean8671513487< LOQM100Concentration and Adsorption of 1-(3-(dodecylamino)propyl)guanidine on BentoniteSampleReplicateSampling Interval0 h 3 h6 hCalc. Conc.[µg/L] At[%]Calc. Conc.[µg/L] At[%]Calc. Conc.[µg/L] At[%]FM11102 0< LOQM 100< LOQM1002 981 4< LOQM 100< LOQM100Mean1042 0< LOQM 100< LOQM100Concentration and Adsorption of 1-(3-(dodecylamino)propyl)guanidineon Glass SurfaceSample RepSampling Interval0 h3 h 24 h 48 h 72 h Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%FS 1743 271209 010220941 8 1008 1 2832 191110 0 963691111 984 4 Mean788 231160 0 993392610 996 3Concentration and Adsorption of 1-dodecyl-1-(3-guanidinopropyl)guanidineon Activated SludgeSampleReplicateSampling Interval0 h 3 h6 hCalc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%]Calc. Conc.[µg/L] At[%]FT1796 227293< LOQM 1002869 157293< LOQM 100Mean833 197293< LOQM 100Concentration and Adsorption of 1-dodecyl-1-(3-guanidinopropyl)guanidineon BentoniteSampleReplicateSampling Interval0 h 3 h6 h Calc. Conc.[µg/L]At[%] Calc. Conc.[µg/L]At[%]Calc. Conc.[µg/L]At[%]FM111560< LOQM100< LOQM 100211150< LOQM100< LOQM 100Mean11360< LOQM100< LOQM 100Concentration and Adsorption of 1-dodecyl-1-(3-guanidinopropyl)guanidine on Glass SurfaceSample Rep Sampling Interval 0 h 3 h24 h 48 h 72 h Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%] Calc. Conc.[µg/L] At[%]FS 1819 201340 0111309497 1083 0 2855 161191 01014 19933 1031 0Mean8371812660106409715 1057 0

Any other information on results incl. tables

The calculation of distribution coefficient is not feasible for the adsorption on activated sludge as well as on bentonitedue to almost complete adsorption on each test system for each component.

For the adsorption on glass surfaces the calculation of distribution coefficients for the component 1-(3-(dodecylamino)propyl)guanidine and 1-dodecyl-1-(3-guanidinopropyl)guanidineis not feasible as well due to no significant adsorption (elimination > 10 %).

For 1-dodecyltetrahydropyrimidin-2(1H)-iminethe distribution coefficient on glass surfaces is:

K_d = 0.00227 L/cm²

For the adsorption on bentonite no extraction was performed due to impracticability. Therefore a calculation of the mass balances was not feasible.

Table:               Calculated Mass Balances for Activated Sludge and Glass Surface

Sample	Calculated Concentration
	Aqueous phase [µg/L]	Extraction [µg/L]	Sum [µg/L]	RR [%]
1-dodecyltetrahydropyrimidin-2(1H)-imine
Activated sludge (FT)	LOQM	1027	1027	101
Glass surface (FS)	479	LOQM	479	47
1-(3-(dodecylamino)propyl)guanidine
Activated sludge (FT)	LOQM	1026	1026	101
Glass surface (FS)	996	LOQM	996	98
1-dodecyl-1-(3-guanidinopropyl)guanidine
Activated sludge (FT)	LOQM	1102	1102	108
Glass surface (FS)	1057	LOQM	1057	104

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The components 1-dodecyltetrahydropyrimidin-2(1H)-imine, 1-(3-(dodecylamino)propyl)guanidine and 1-dodecyl-1-(3-guanidinopropyl)guanidine of the test item showed a high tendency to adsorb on activated sludge and bentonite with adsorption > 90 % after 6 h and 3 h, respectively.The adsorbed analytes could be successfully extracted from activated sludge, leading to a valid mass balance. During method development it was proven, that the analysed components of the test item adsorb irreversible to bentonite and therefore no mass balance could be achieved.Adsorption on glass surfaces was only observed for the component 1 dodecyltetrahydropyrimidin-2(1H)-imine with a degree of adsorption (At) of 53 %, corresponding to a distribution coefficient of 0.00227 L/cm². Extraction from the glass surface was not successful, resulting in a mass balance of 47 % for this component.For the other two components, 1-(3-(dodecylamino)propyl)guanidine and 1-dodecyl-1-(3-guanidinopropyl)guanidine, no adsorption was observed.With respect to the test conditions as well as to the successful mass balances, abiotic degradation, e.g. hydrolysis or photolysis, can be negated.The degree of adsorption of the reference item was determined to be 98 % after 24 h confirming the validity of the study design.

Executive summary:

The adsorption behaviour of the test item on activated sludge, bentonite and glass surfaces was determined according to ISO 18749:2004(E) from 2012‑07‑27 to 2012‑12‑03 at Dr.U.Noack-Laboratorien, 31157 , .

Analyses were performed via LC-MS/MS on a reversed phase column using an external standard. The analytical method for determination of the test item was validated and tested with satisfactory results in regard to linearity, accuracy, precision and specificity.

The components 1-dodecyltetrahydropyrimidin-2(1H)-imine, 1-(3-(dodecylamino)propyl)guanidineand 1-dodecyl-1-(3-guanidinopropyl)guanidineof the test item were selected as representative analytes and showed a high tendency to adsorb on activated sludge and bentonite with an adsorption > 90 % after 6 h and 3 h, respectively.
The adsorbed matter on activated sludge could successfully be extracted, confirmed by the valid mass balance of 101 – 108 %. During method development it was proven, that the analysed components of the test item adsorb irreversible to bentonite and therefore no mass balance could be achieved.

Adsorption on glass surfaces was only observed for the component 1‑dodecyltetrahydropyrimidin-2(1H)-iminewith a degree of adsorption (A_t) of 53 %, corresponding to a distribution coefficient of 0.00227 L/cm². Extraction from the glass surface was not successful, resulting in a mass balance of 47 % for this component.
For the other two components, 1-(3-(dodecylamino)propyl)guanidineand 1-dodecyl-1-(3-guanidinopropyl)guanidine, no adsorption was detected.

With respect to the test conditions as well as to the successful mass balances, abiotic degradation, e.g. hydrolysis or photolysis, can be negated.

The degree of adsorption of the reference item was determined to be 98 % after 24 h confirming the validity of the study design.

The adsorption of test item was determined to be:

·               > 90 % on activated sludge after 6 h for each analysed component

·               > 90 % on bentonite after 3 h for each analysed component

·               47 % on glass surfaces after 72 h for the component
1-dodecyltetrahydropyrimidin-2(1H)-imine
with a distribution coefficient of K_d = 0.00227 L/cm²

·                5 % on glass surfaces after 72 h for the components
1-(3-(dodecylamino)propyl)guanidineand 1-dodecyl-1-(3-guanidinopropyl)guanidine