ASTM D-6400 - worldcentric.com · ASTM D-6400 Determining Aerobic Biodegradation of Plastic Materials Under Controlled Composting Conditions Tested By: Soil Control Lab 42 Hangar

ASTM D-6400

Determining Aerobic Biodegradation of Plastic

Materials Under Controlled Composting

Conditions

Tested By:

Soil Control Lab

42 Hangar Way

Watsonville, CA 95076

(831) 724-5422

www.compostlab.com

Tested For:

Aseem Das

World Centric Corporation

2121 Strunton Court

Palo Alto, CA 94306

Results:

SCL 9070001

Account No.:

9070001-1-09

Batch

42 Hangar Way Jul. 09

Watsonville CA 95076 CODE: ASTM 6400

www.compostlab.com

Aseem

World Centric

2121 Strunton Court

Palo Alto CA 94306

Date Received: 30 Jun. -09

Sample Id.: Wheat Straw Trays

Sample id. Number 9070001

ASTM D6400 - BPI -Preliminary Report

Total Carbon 41.0 percent dry weight

Ash 5.4 percent dry weight

Thickness of Material 0.516 mm

BioBased (D6866) 93 +/- 1 percent

BioBased (ASTM D6868) Info.

http://www.soybiobased.org/resources/meeting012904_ASTM_Briefing.doc

Heavy Metal Concentration

Constituents Sample U.S. Canadian

mg/kg dry wt. Limits Limits

Arsenic As 0.72 22 38

Cadmium Cd 0.26 20 10

Copper Cu 4.89 750 NA

Lead Pb 3.00 150 250

Mercury Hg < 1 9 3

Nickel Ni 0.60 210 90

Selenium Se < 1 50 7

Zinc Zn 19.3 1400 925

Cobalt Co 0.28 NA 75

Chromium Cr 0.89 NA NA

Molybdenu Mo 0.51 NA 10

Fluorine F < 1 100 100

The above analysis for heavy metals was performed in house by SCL using the following methods:

EPA Method 207 (ICP) for Cd, Cu, Pb, Ni, Zn, Co, Cr and Mo

EPA Method 208 (ICPMS) for As, Se

EPA Method 245.7 (cold vapor) Hg

EPA Method 300.1 (IC) for F

Analyst: Frank Shields Page 2

Soil Control Lab

42 Hangar Way

Watsonville, CA 95076

Description of Test Procedure

Equipment:

Two controlled temperature ovens that have capacities to hold 36 test chambers each for a total of

72 Test chambers; Each cylindrical chamber has a capacity of seven and one half liters.

The containers are sectioned into two parts by use of a porous pad so that the top section has a

volume of six liters. One liter of water is placed in the bottom section and the test material in the top.

Carbon dioxide free air is bubbled through the water to water saturate the air and then through the

porous pad into the main test chamber then out of the the top. The air exiting the test chambers

is plumbed to a solenoid valve which is controlled so that every two hours the air is diverted for two

minutes. This diverted air flows into one liter adsorption units containing a known volume of one normal

sodium hydroxide to adsorb the carbon dioxide produced in the test chambers.

Test Procedure

A suitable three month old stable compost from Monterey District composting facility is used for the

Inoculum of this test. The material is sieved through 9.5 mm (0.371 inches) and then mixed.

A known weight of inoculum is taken so that it is equivalent to 600 grams of dry weight, ammonium

chloride is added so that the C/N ratio is less than 15 plus an appropriate volume of water to adjust

the moisture content to 55%. The inoculum is mixed in a mixer and then transferred to a test .

chamber. Positive and negative controls are established by adding 100 grams of cellulose powder

and polyethylene respectively to the inoculums Finally 100 grams of the test material which has .

been reduced to a size of less than 2cm. is added to the inoculum formula. The above mixtures

are set up in triplicate to give a total of 12 test containers.

The test chambers are placed in the temperature controlled oven which is set to 58 degrees

centigrade. The air is then connected and adjusted so that the flow rate is between 150 and 200

milliliters per minute. After 3 days the adsorption units are removed so that the

amount of carbon dioxide produced can be determined by titrating to pH 8.5 with 0.5 normal

hydrochloric acid after adding barium chloride to precipitate the carbonates formed by the

carbon dioxide produced in the test chambers.

The adsorption units are then replaced and the same procedure is performed for a furthers 4 days.

Then on a weekly basis the chambers are removed from the oven opened to replace whatever

volume of water that has been used and to mix the contents of the chambers. After returning the

chambers to the oven the air is connected and the carbon dioxide adsorption procedure allowed to

proceed for 7 days. The testing is allowed to proceed for a total of 12 weeks or until the amount

of carbon converted to carbon dioxide in the test containers has ceased.

The cumulative amount of carbon dioxide is calculated for each chamber so that the amount of

carbon converted can be calculated for the test sample and the positive and negative controls.

The material in the chambers is removed and sieved through 2mm sieve to establish the percentage

of disintegration. The material from the triplicate test chambers that pass through the sieve is then

mixed in a blender and subjected to the plant phytotoxicity study. Page 2

Work Order Number

SCL - 9070001

Soil Control Lab Batch

42 Hangar Way Jun 2009

Watsonville CA 95076 CODE: ASTM 6400

www.compostlab.com

Aseem

World Centric

2121 Strunton Court

Palo Alto CA 94306

Date Received: 30 Jun. 09


Sample id. Number 1 9070001

ASTM D6400 Test Package for BPI Certification

Description: Wheat Straw Trays cardboard like

Start Date: 06 Jul. 09 End Date: 23 Sept. 09

Sample Pretreatment: Cut into 2 cm squares.

Inoculum Pretreatment: Pre-screened through 9.5mm sieve and stored at 4 deg. C until needed.

Analysis of inoculum: Units Units

Resp. Rate in Unit: 78 *See below Phosphorus (P) 3800 mg/kg dry

pH value: 7.25 units Potassium (K) 11000 mg/kg dry

Moisture: 36.6 percent dry E Conductivity (EC5) 7.80 mmhos/cm

Volatile Solids: 45.0 percent dry AgIndex Ratio 6 Ratio

Total Organic Carbon 26 percent dry Respiration Rate 1 **See below

Total Nitrogen (N) 1.4 percent dry Biological Avail. C (BAC 1.1 **See below

Ammonia Nitrogen 180 mg/kg dry Maturity (germination) 100 % of control

Nitrate Nitrogen 440 mg/kg dry Maturity (vigor) 100 % of control

Organic Nitrogen 1.3 percent dry Metals Pass Pass/Fail

Carbon/Nitrogen (C/N) 18.6 Ratio Collected then screened and stored @4 deg. C

*CO2-C per gram VS in the first ten days ** mg CO2-C/g OM/day

Set Up:

Tweleve reactors were prepared as follows:

Inoculum Blank: (3 reps.):

946 g compost, 254 mls water and 8g NH4Cl.

Positive Control (3 reps.):

946 g compost, 354 mls water,8 g NH4Cl and100 g powdered cellulose.

Negative Control: (3 reps.)

946 g compost, 354 mls water,8 g NH4Cl, and 100 g cut sheet polyehylene.

Sample (3 Reps.):

946 g compost, 354 mls water, 8 g NH4Cl, and 100 g prepared sample.

Mixes prepared to have the final mix 50 percent moisture and a C/N ratio less than 30.

Page 3

Soil Control Lab Work Order Number SCL - 9070001

42 Hangar Way Batch Jun 2009

Watsonville, CA 95076 Code: ASTM 6400


Data Summary

Pass / Fail Readings during first 12 weeks (84 days) of the test.

Day 10 The suggested carbon dioxide production of the inoculum is between 50 and 150 mg

per grams volatile solids in the first ten days. Actual value = 78

PASS Day 45 Cellulose must biodegrade greater than 70%. Actual value is 77.1.

PASS Standard deviation of the positive reference must be less than 20%. Actual value is 11.53

Day 84 The degree of disintigation in 12 weeks was visibly greater than 90 percent.

The degree of disintegration calculated to be approximately 95 percent.

The reactors containing the test material showed signs of decomposition at day 24

as the appearance became covered in fungus and soft. After day 45 the material

started to disintegrate into smaller pieces.

Reading at the end of test:

Day 80 End of Test

Weight of container contents reported dry weight:

Start (g) End (g) Loss (g) Start (g) End (g) Loss (g)

inoculcum 602 530 72 Pos. Control 702 562 140



Neg Control 702 640 62 Sample 702 562 140



PASS The pH value at the end of the test was greater than seven. Actual value = 7.6

PASS The temperature range of the incubator was maintained at 58 deg. C +/- 2 deg C.

The degree of biodegradation determined to be 95% after 80 days

PASS The concentrations of heavy metals was less than the published limits.

There was satisfactory appearance, odor and moisture content of all chambers .

throughout the test.

Phytotoxicity

PASS % Germ. Cucumber The phytotoxicity test for two plant species determined using percent germination

PASS % Germ. Turf Mix and percent vigor PASS % Germ.. Test Method used is TMECC 05.05-A for phytotoxicity.

PASS % Vigor" Cucumber Closed containers to concentrate any volatile and prevent moisture loss is used to

PASS % Vigor Turf Mix keep any toxins present close to the plant. Fast growing, salt tolerant plants are chosen

for the test specimen to aquir lower detection limits. Mixes diluted with vermiculite until

positive control produced healthy plants. Calculations as percent of positive control.

Results: Germination % STDEV Vigor % STDEV

Cucumber 90 13.9 99 2.0

Turf Mix 98 5.8 109 2.7

PASS Single Polymer Carbon converted from cellulose is 79.5 %. and from test material is 75.5%

PASS Multi Polymer Carbon converted from test material using cellulose as positive control is 95%.

The standard deviation and 95% confidence inteval for cellulose was 10.53 and 11.92 res

The standard deviation and 95% confidence inteval for sample was 2.75 and 3.11 resp.

Page 4




Day Reactor 1 Reactor 2 Reactor 3 Day Reactor 1 Reactor 2 Reactor 3

0 0 0 0 0 0 0 0

3 8.00 7.87 8.57 3 8.00 7.87 8.57

7 9.16 7.94 6.91 7 17.16 15.81 15.48

10 5.36 5.44 4.23 10 22.52 21.25 19.71

14 6.58 5.65 5.67 14 29.10 26.90 25.38

17 5.39 5.07 4.55 17 34.49 31.97 29.93

21 6.56 5.63 5.50 21 41.05 37.60 35.43

24 4.53 5.13 4.97 24 45.58 42.73 40.40

31 7.18 5.62 6.76 31 52.76 48.35 47.16

38 7.24 6.00 6.03 38 60.00 54.35 53.19

45 6.35 6.87 5.57 45 66.35 61.22 58.76

52 6.65 6.71 5.50 52 73.00 67.93 64.26

59 5.62 6.15 4.94 59 78.62 74.08 69.20

66 6.85 6.42 5.13 66 85.47 80.50 74.33

73 6.98 7.03 4.67 73 92.45 87.53 79.00

80 5.18 5.76 4.12 80 97.63 93.29 83.12

% Standard 95% Conf.

Day Average STDV % STDV Population Error Limit Coef.of.var

45 62 3.87 6.23 3 2.24 4.38 0.0623

80 91 7.45 8.15 3 4.30 8.43 0.0815

Page 5

Carbon Dioxide Production from Inoculum

Measured Production (gm) Cumulative Production (gm)

Cumulative Production of CO2 by Inoculums

0

20

40

60

80

100

120

0 20 40 60 80 100

day

CO

2 p

rod

ucti

on

(g

m)

Reactor 1

Reactor 2

Reactor 3





0 0 0 0 0 0 0 0

3 8.19 8.26 8.03 3 8.19 8.26 8.03

7 7.81 7.72 7.94 7 16.00 15.98 15.97

10 5.44 5.34 4.87 10 21.44 21.32 20.84

14 6.25 5.85 6.84 14 27.69 27.17 27.68

17 4.61 4.84 5.24 17 32.30 32.01 32.92

21 5.80 5.41 6.47 21 38.10 37.42 39.39

24 4.53 5.13 4.97 24 42.63 42.55 44.36

31 7.18 5.62 6.76 31 49.81 48.17 51.12

38 7.24 12.41 6.03 38 57.05 60.58 57.15

45 6.35 6.87 5.57 45 63.40 67.45 62.72

52 6.65 6.71 5.50 52 70.05 74.16 68.22

59 5.62 6.15 4.94 59 75.67 80.31 73.16

66 6.85 6.42 5.13 66 82.52 86.73 78.29

73 6.98 7.03 4.67 73 89.50 93.76 82.96

80 5.18 5.76 4.12 80 94.68 99.52 87.08



45 65 2.56 3.96 3 1.48 2.89 0.0396

80 94 6.27 6.69 3 3.62 7.10 0.0669

Page 6

Carbon Dioxide Production from Inoculum + Polyethylene

Cumulative Production (gm)Measured Production (gm)

Cumulative Production of CO2 by Inoculum +

Polyethylene

0

20

40

60

80

100

120

0 20 40 60 80 100

day

CO

2 p

roduction (

gm

)

Reactor 1

Reactor 2

Reactor 3





0 0 0 0 0 0 0 0

3 42.31 33.11 26.59 3 42.31 33.11 26.59

7 47.03 48.96 49.08 7 89.34 82.07 75.67

10 17.38 15.37 20.69 10 106.72 97.44 96.36

14 15.68 15.98 16.00 14 122.40 113.42 112.36

17 11.08 9.71 10.20 17 133.48 123.13 122.56

21 12.10 10.98 11.44 21 145.58 134.11 134.00

24 8.73 7.51 7.55 24 154.31 141.62 141.55

31 13.91 13.24 12.17 31 168.22 154.86 153.72

38 13.35 10.36 9.73 38 181.57 165.22 163.45

45 12.00 9.13 9.46 45 193.57 174.35 172.91

52 8.40 7.94 8.19 52 201.97 182.29 181.10

59 6.46 6.75 6.92 59 208.43 189.04 188.02

66 5.25 6.50 6.88 66 213.68 195.54 194.90

73 6.49 6.64 6.22 73 220.17 202.18 201.12

80 5.19 5.47 5.50 80 225.36 207.65 206.62



45 180 11.53 6.40 3 6.66 13.05 0.0640

80 213 10.53 4.94 3 6.08 11.92 0.0494

Page 7

Cumulative Production (gm)

Carbon Dioxide Production of Inoculum + Cellulose

Measured Production (gm)

Cumulative CO2 Production of Cellulose + Inoculum

0

50

100

150

200

250

0 20 40 60 80 100

day

CO

2 p

rod

ucti

on

(g

m)

Reactor 1

Reactor 2

Reactor 3





0 0 0 0 0 0 0 0

3 16.88 16.07 19.75 3 17 16 20

7 24.09 24.40 35.48 7 41 40 55

10 13.63 17.91 23.24 10 55 58 78

14 18.23 20.56 19.25 14 73 79 98

17 14.93 22.28 13.83 17 88 101 112

21 17.51 21.12 12.76 21 105 122 124

24 9.90 9.78 8.69 24 115 132 133

31 16.91 15.23 14.36 31 132 147 147

38 17.04 12.41 10.32 38 149 160 158

45 14.39 9.42 8.57 45 164 169 166

52 10.80 7.86 8.14 52 174 177 174

59 8.95 7.24 8.93 59 183 184 183

66 8.31 7.69 6.40 66 192 192 190

73 8.39 6.78 6.48 73 200 199 196

80 7.43 6.52 5.74 80 207 205 202



45 166 2.84 1.70 3 1.64 3.21 0.0170

80 205 2.75 1.34 3 1.59 3.11 0.0134

Page 8

Cumulative Production (gm)Measured Production (gm)

Carbon Dioxide Production of Inoculum + Test Material

Cumulative Production of CO2 of Sample + Inoculum

0

50

100

150

200

250

0 10 20 30 40 50 60 70 80 90

day

CO

2 p

rod

ucti

on

(g

m)

Reactor 1

Reactor 2

Reactor 3




Day Cellulose Sample Polyethylene Inoculum

0 0 0 0 0

3 34 18 8 8

7 82 46 16 16

10 100 64 21 21

14 116 83 28 27

17 126 100 32 32

21 138 117 38 38

24 146 127 43 43

31 159 142 50 49

38 170 156 58 56

45 180 166 65 62

52 188 175 71 68

59 195 184 76 74

66 201 191 83 80

73 208 198 89 86

80 213 205 94 91

Page 9

Mean Cumulative Carbon Dioxide Production in Reactors

Carbon Dioxide Production (grams)

Mean Cumulative Carbon Dioxide Production in Reactors

0

50

100

150

200

250

0 20 40 60 80 100

day

CO

2 p

roduction (

gm

)

Cellulose

Sample

Polyethylene

Inoculum




Cellulose Sample Polyethylene

Day gm gm gm Day Cellulose Sample Polyethylene

0 0 0 0 0 0 0 0

3 7.05 2.57 0.00 3 17 6 0.00

7 18.06 8.02 -0.05 7 43 20 -0.05

10 21.55 11.63 0.01 10 52 28 0.01

14 24.25 15.28 0.11 14 58 37 0.12

17 25.71 18.56 0.08 17 62 45 0.09

21 27.24 21.62 0.08 21 65 53 0.09

24 28.07 22.87 0.08 24 67 56 0.09

31 29.87 25.32 0.08 31 71 62 0.09

38 31.15 27.18 0.66 38 75 66 0.77

45 32.23 28.42 0.66 45 77 69 0.77

52 32.74 29.14 0.66 52 78 71 0.77

59 33.05 29.91 0.66 59 79 73 0.77

66 33.07 30.27 0.66 66 79 74 0.77

73 33.14 30.54 0.66 73 79 75 0.77

80 33.24 30.96 0.66 80 80 76 0.77

Total Carbon

Cellulose Sample Polyethylene

41.80 40.99 85.60 Carbon (g) / 100 g

Page 10

Cumulative Added Carbon Converted to CO2 in Reactors (mean of three)

Percent Converted

Percent Carbon Converted to CO2

-10

0

10

20

30

40

50

60

70

80

90

0 20 40 60 80 100

day

% C

convert

ed

Cellulose

Sample

Polyethylene




Amount of Carbon Converted to Carbon Dioxide

Work Order Number SCL - 9070001

Day % converted

0 0

3 37

7 45

10 55

14 64

17 74

21 81

24 83

31 86

38 89

45 90

52 91

59 92

66 93

73 94

80 95

Page 11

(as compared to cellulose as positive control)

Percent Carbon Converted Using Cellulose as Control

0

20

40

60

80

100

0 20 40 60 80 100

day

% C

convert

ed

% converted




Day Cellulose Sample Polyethylene Inoculum

0 0 0 0 0

3 8.24 4.26 1.98 1.97

7 10.27 5.21 2.32 1.64

10 6.99 3.95 1.70 1.32

14 8.44 4.75 2.57 1.41

17 7.57 3.81 1.94 1.35

21 4.58 3.94 1.46 1.58

24 4.86 4.99 1.28 1.29

31 3.62 3.20 1.31 1.10

38 3.42 3.24 1.67 1.05

45 2.80 2.58 1.15 0.87

52 2.56 2.43 1.29 0.85

59 2.21 2.15 1.41 0.87

66 2.21 2.08 1.27 0.87

73 2.07 1.58 0.97 0.86

80 2.13 1.66 1.36 0.91

Page 12

Oxygen Consumption (grams per day)

Mean Oxygen Consumption in Reactors

Oxygen Consumption of Reactor

0

2

4

6

8

10

12

0 20 40 60 80 100

Day

gra

ms p

er

day

Cellulose

Sample

Polyethylene

Inoculum




Plant Growth Study

Cucumber

Germination Height

Init. Dilution.(sample:vermiculite) v/v 10 : 1 10 : 1

# Seeds per pot 15 15

# of replicates 3 3

Percent Average

% STDEV (Less than 20) 14 2

% of Control (Greater than 90) 90 99

Pass/Fail PASS PASS

Turf Mix

Germination Height

Init. Dilution.(sample:vermiculite) v/v 10 : 1 10 : 1

# Seeds per pot 30 30

# of replicates 3 3

Percent Average

% STDEV (Less than 20) 6 3

% of Control (Greater than 90) 98 109

Pass/Fail PASS PASS

Description of Test:

Germination and vigor of the seeds planted were reported against the

positive control and reported as percent of the positive control. The pots used were

cups with clear plastic cover. This to hold in moisture reducing the need to water and

possible leaching of phytotoxins and to hold and concentrate any volitile toxins around

the growing plant. Because compost is not a good potting soil due to excessive salts

and nutrients several dilutions of the two mixes (sample and positive controll -cellulose)

were made. Triplicate of each of the dilutions and another set of plain vermiculite

were all seeded. Vermiculite was used as a negative control to make sure the dilution

media was suitable. Test procedure is taken from the TMECC for the testing of phytotoxins .

in compost materials. Also adapting interpretation from OECD "Terrestrial Plant Test".

Plants were monitored during the two weeks growth.

The most concentrated dilution of the positive control that produced the best growth was

used when interpretating the results.

Page 13

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ASTM D-6400 - worldcentric.com · ASTM D-6400 Determining Aerobic Biodegradation of Plastic Materials Under Controlled Composting Conditions Tested By: Soil Control Lab 42 Hangar