Dihydronicotinamide-adenine dinucleotide, disodium salt

Partition coefficient

Administrative data

Endpoint:

partition coefficient

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 - 13 September

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Reference

Endpoint:

water solubility

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 - 25 August 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 105 (Water Solubility)

Version / remarks:

1995

Deviations:

yes

Remarks:

see principles of method if other than guideline

Qualifier:

according to guideline

Guideline:

EU Method A.6 (Water Solubility)

Version / remarks:

Regulation (EC) No 440/2008

Deviations:

yes

Remarks:

see principles of method if other than guideline

Principles of method if other than guideline:

Due to very high water solubility of the test item, estimated in a preliminary test, a modified TG OECD 105 design was conducted. Instead of equilibrating a large amount of test item with water by stirring over several days and subsequently separating the test item (about five times in excess) from the saturated solution, water was added in a stepwise procedure to a defined amount of test item until complete dissolution occurred.

GLP compliance:

yes (incl. QA statement)

Type of method:

flask method

Key result

Water solubility:

>= 712 g/L

Conc. based on:

test mat. (dissolved fraction)

Temp.:

22.5 °C

pH:

7

Details on results:

Preliminary test
Approximately 0.1 g of the test item were completely dissolved in 0.2 mL of ultra-pure water indicating a water solubility of about 500-1000 g/L.

Main test
The results are given in the table in the next section. The water solubility was determined as the mean of the triplicate test assays analysed with HPLC. As this method does not ensure complete saturation of the solution, it is given as a minimum value in mass per volume of solution. The test is satisfactory, if the concentrations of the three vessels do not differ by more than 15%.

HPLC determinations:

Test vessel	pH	Analysed concentration A (g/L)	Analysed concentration B (g/L)	Mean (g/L)
No. 1	7	706	708	707
No. 2	7	720	718	719
No. 3	7	711	710	710
			Overall Mean	712

The concentrations of the three vessels differed by 2% (validity criterion given by OECD is met).

Further remarks

Based on the test item amount used (ca. 1 g) and the volume of ultra-pure water applied (0.8 ml) to achieve complete dissolution, a theoretical solubility of ca. =1250 g/l results, which is significantly higher than the water solubility determined in this test. It has to be considered in this case that the final volume of the test item solution is most likely higher than just the 0.8 ml of the ultra-pure water added, due to the very high test item concentrations.

Conclusions:

The water solubility of beta-NADH, disodium salt (CAS no. 606-68-8) was determined according to a modified TG OECD 105 design.
The water solubility of the test item at 22.5°C was >=712 g/L. The pH of the aqueous samples was 7.

Executive summary:

The water solubility of beta-NADH, disodium salt (CAS no. 606-68-8) was determined according to a modified TG OECD 105 design.

To this end, increasing volumes of water were added in a stepwise procedure, to approximately 1 g of the test item at 22 ± 2 °C until complete dissolution, which occurred after addition of 0.8 mL of water. The concentration of test item was subsequently determined by HPLC with UV detection.

The determined water solubility of beta-NADH, disodium salt is >=712 g/L at pH 7 and at 22.5 °C.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 107 (Partition Coefficient (n-octanol / water), Shake Flask Method)

Version / remarks:

1995

Deviations:

yes

Remarks:

see principles of method if other than guideline

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method A.8 (Partition Coefficient - Shake Flask Method)

Version / remarks:

Regulation (EC) No 440/2008

Deviations:

yes

Remarks:

see principles of method if other than guideline

Principles of method if other than guideline:

The test was not performed with the non-charged species, as given in the guideline, as the amount of non-charged species in the environmentally relevant pH range of 5 to 9 is negligible. Instead, the single negatively charged species dominates.

GLP compliance:

yes (incl. QA statement)

Type of method:

flask method

Partition coefficient type:

octanol-water

Test material

Test material information

Constituent 1

Reference substance name:

Dihydronicotinamide-adenine dinucleotide, disodium salt

EC Number:

210-123-3

EC Name:

Dihydronicotinamide-adenine dinucleotide, disodium salt

Cas Number:

606-68-8

Molecular formula:

C21H27N7O14P2.2Na

IUPAC Name:

disodium [(2R,3S,4R,5R)-5-(3-carbamoyl-1,4-dihydropyridin-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl ({[(2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphonato}oxy)phosphonate

impurity 1

Reference substance name:

Dihydronicotinamide-adenine dinucleotide

EC Number:

200-393-0

EC Name:

Dihydronicotinamide-adenine dinucleotide

Cas Number:

58-68-4

Molecular formula:

C21H29N7O14P2

IUPAC Name:

[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2R,3S,4R,5R)-5-(3-carbamoyl-4H-pyridin-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl hydrogen phosphate

impurity 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

Oxidane

Test material form:

solid: crystalline

Remarks:

crystaline powder

Details on test material:

Batch No.: 18243100
Storage conditions: Store at -15 to -25°C,
Handling: Keep frozen on dry ice.

Study design

Analytical method:

high-performance liquid chromatography

Results and discussion

Partition coefficient

Key result

Type:

log Pow

Partition coefficient:

< -1.96

Temp.:

21 °C

pH:

7

Details on results:

Calculation method
The log POW of the test item molecule was estimated to be -9.00, as reported in EPI Suite™ v4.11, KOWWIN v1.68. This predicted value is out of the range of the EPI Suite training set (log POW values = -5). Nevertheless, it may be used as indication that the log Pow of the test item is very low.

Flask method
The determined log POW values were all < -1.96 for all three volume ratios and all replicates. The mass balance recovery rates in the range of 94% to 96% seem to indicate a slight test item loss. However, the recovery rate for the aqueous, buffered stock solution, which was used for the preparation of the test assays, was in the same range with 95%. This confirms the initial dosage as well as the determined test item concentrations in the 1-octanol phase, which were all below the LOQ of 67.9 mg/L.
The determined log POW value of beta-NADH, disodium salt (CAS no. 606-68-8) at pH 7.0 is < -1.96 with a standard deviation of ± 0.002.

The log Pow of the test item is given as the mean value of all measurements. Detailed results are given in the table in the field 'Any other information on results incl. tables'.

Any other information on results incl. tables

Ratio n-octanol:water (v:v)	Analysed concentration n-octanol (mg/L)	Analysed concentration water (mg/L)		Pow	log Pow	pH
1:4	<67.9	6196	<0.0110		<-1.96	7.0
1:4	<67.9	6209	<0.0109		<-1.96	7.0
1:9	<67.9	6152	<0.0110		<-1.96	7.0
1:9	<67.9	6153	<0.0110		<-1.96	7.0
1:19	<67.9	6149	<0.0110		<-1.96	7.0
1:19	<67.9	6121	<0.0111		<-1.96	7.0
Mean					<-1.96

Further remarks:

The result obtained in this study confirms that the log POW of beta-NADH, disodium salt is quite low at a pH in the environmentally relevant range of pH 5 to 7. This is in line with a determined water solubility of >= 712 g/L at pH 7 and at 22.5 °C (see cross-ref to water solubility).

Applicant's summary and conclusion

Conclusions:

The shake flask method according to TG OECD 107 was applied for the determination of the log Pow. The determined log POW value of beta-NADH, disodium salt (CAS no. 606-68-8) at pH 7.0 is <-1.96 with a standard deviation of ± 0.002.

Executive summary:

The log POW of the test item beta-NADH, disodium salt (CAS no. 606-68-8) was determined according to TG OECD 107.

Aqueous phosphate buffer at pH 7.0, 1-octanol and the test item were equilibrated in test vessels by shaking, applying three different volume ratios of the two solvents. Subsequently, the test item concentrations were determined in both solvent phases.

The determined log POW values were all <-1.96 for all three volume ratios and all replicates.

The determined log POW value of beta-NADH, disodium salt at pH 7.0 is <-1.96 with a standard deviation of ± 0.002.

The validity criterion was fulfilled.