SPSFAM ERP

Table 8. Reproducibility. For each sample, duplicate extractions were processed and applied to the assay on 4 separate occasions by 2 independent analysts.

Interday: Fructan, % (w/w), mean, ±2 SD, (%RSD r )

Fructan, % (w/w), mean ±2 SD, (%CV)

Sample

A1 n1

A1 n2

A1 n3

A1 n4

A2 n1

A2 n2

A2 n3

A2 n4

99.8 ± 4.1 98.2 ± 1.2 98.3 ± 0 97.2 ± 1.5 98.9 ± 0.4 104.8 ± 19.5 103.3 ± 2 101.9 ± 1.7

100.3 ±7.4

Raftiline Inulin (native chicory fructan)

2.04

0.61

0.00

0.77

0.19

9.32

0.98

0.82

3.70

97.5 ± 1.6 98.3 ± 2

99.2 ± 1 98.6 ± 0.9 97 ± 0.2

97.9 ± 2.3 99.8 ± 0.1 101.3 ± 1

98.7 ±2.8

P-­‐LEVAN #171114

0.83

1.01

0.50

0.45

0.09

1.17

0.07

0.49

1.44

85.7 ± 3.5 86.4 ± 2

86.8 ± 3 83.2 ± 1.5 78.7 ± 0.9 80.4 ± 4.3 84.3 ± 2.5 80.9 ± 1.1

83.3 ±6.2

Frutafit Agave fructan

2.07

1.17

1.75

0.91

0.55

2.66

1.49

0.66

3.69

14.4 ± 0.8 14.3 ± 0.1 14.8 ± 0.4 14.8 ± 0.5 15.6 ± 0.2 15.9 ± 1.1 15.2 ± 1 14.9 ± 0.8

15 ±1.2

Timothy grass #171201

2.93

0.50

1.50

1.61

0.51

3.58

3.20

2.84

4.01

14.4 ± 0.2 14 ± 1.4 14.7 ± 0.7 14.3 ± 0.6 12.7 ± 0.6

-­‐ -­‐

14.6 ± 2.6 14.2 ± 2.3

14.1 ±1.6

Barley Max

0.60

4.92

2.46

2.27

2.51

8.84

7.94

5.77

11 ± 0.4 10.8 ± 0.3 10.4 ± 0.6 10.4 ± 0.1 10 ± 0.1

10.3 ± 0.2 10.3 ± 0.5 10.4 ± 0.3

10.4 ±0.7

Rye Grass

1.73 2 ± 0 0.78 0.32 1 ± 0 1.73

1.25 2 ± 0 0.31

2.98

0.46

0.57

0.74

2.27

1.63

3.25

2 ± 0.1

1.9 ± 0.1

1.9 ± 0.1

2 ± 0.1

2.3 ± 0.1 2.1 ± 0.4

2.01 ±0.26

Brett Brother swine feed

3.05

2.64

3.11

2.33

1.69

9.46

6.36

1.7 ± 0

1.7 ± 0.1

1.7 ± 0.1

1.5 ± 0.1

1.4 ± 0

1.5 ± 0.1

1.7 ± 0.3

1.7 ± 0

1.6 ±0.2

Barley Flour #60301a

2.80

3.04 1 ± 0 0.16

2.78 1 ± 0 0.78

1.12

1.91

8.23

0.50

5.79

1 ± 0.2

0.9 ± 0

0.9 ± 0

1 ± 0.1

1 ± 0.1

0.98 ±0.1

Purina GOCat dry cat food

9.18

1.54

1.17

4.55

3.65

4.84

0.2 ± 0

0.2 ± 0

0.2 ± 0

0.2 ± 0

0.2 ± 0

0.2 ± 0

0.2 ± 0

0.2 ± 0.1

0.19 ±0.03

Top spec alfalfa

4.20

3.82

2.91

8.53

8.53

9.37

3.48

20.75

8.47

A1 = Analyst 1 A2 = Analyst 2

6.2.8 Stability The Fructan Assay Kit (K-FRUC) as formulated by Megazyme comes with a two- year stability guarantee. If the kit components are prepared and stored as described in the kit booklet, they also are stable for at least 2 years. Individual kit components may have longer stability guarantees: this information is available on the component label as the expiry date. Regular quality control testing is performed within the Megazyme QC laboratory. As the Fructan Assay Kit has been commercially available for a number of years (excluding the endo -levanase enzyme which is a new addition), the historical data available from the Megazyme Quality Control laboratory shows that the stability of each component does not vary from batch to batch (data not shown). 6.2.9 Robustness The effect of the temperature of incubation with the sucrase/ β -amylase/pullulanase mixture, the fructanase mixture and sodium borohydride was evaluated by performing incubations at both 30 o C and 40 o C. With the sucrase/ β -amylase/pullulanase mixture and the fructanase mixture very similar results were obtained at both 30 o C and 40 o C.

25