Absorbances at 600nm

Lipopeptides standard curves

protocol 1 7.059
0.25

y = 0.1867x - 0.0026
R = 0.9884

0.2

0.15
0.1
0.05
0
-0.05

0.2

0.4

0.6

0.8

1.2

Concentration of lipopeptides in mg/ml

Lipopeptides standard curves

protocol 2 1.984
Absorbances at 600nm

0.2
y = 0.1722x + 0.0007
R = 0.9983

0.15
0.1
0.05
0
0

0.2

0.4

0.6

0.8

Concentration of lipopeptides in mg/ml

1.2

0.0508g ofcrude lipopeptides dissolved in autoclaved and 0.2micron filtered TSB+ YE+Mn medium at pH 7.059
Gently vortexed to mix the samples and appropriate volumes of solutions were mixed, gently homogenized and incubated at 4 C.
After 30minutes, the solutions were again mixed and readings were noted at 600nm using the blank.
For 1.984, 3 replicates were used, while for that of 7.059 only one replicate was used
All the reactions were performed in 1ml cuvettes.
Samples

1.984
Blank
Standard 1
Standard 2
Standard 3
Standard 4
Standard 5

Volume of
lipopeptides
stock
ml
0
0.2
0.4
0.6
0.8
1

Volume of TSB+ YE +
Mn stock (pH 1.984 or
7.059)
ml
1
0.8
0.6
0.4
0.2
0

0
0.2
0.4
0.6
0.8
1

1
0.8
0.6
0.4
0.2
0

Volume Concentartion
Absorbances at
Absorbances at
of 6N oflipopeptides
600nm 1.1ml
600nm1.0ml
HCl
ml
mg/ml
1
2
3
1
2
3
0.1
0
0
0.001 0.001
0
0.0011 0.0011
0.1
0.2032
0.031 0.032 0.032 0.0341 0.0352 0.0352
0.1
0.4064
0.064 0.061 0.062 0.0704 0.0671 0.0682
0.1
0.6096
0.103 0.096 0.098 0.1133 0.1056 0.1078
0.1
0.8128
0.134 0.131 0.127 0.1474 0.1441 0.1397
0.1
1.016
0.166 0.152 0.152 0.1826 0.1672 0.1672

Averages

0.000733
0.034833
0.068567
0.1089
0.143733
0.172333

7.059
Blank
Standard 1
Standard 2
Standard 3
Standard 4
Standard 5

0.1
0.1
0.1
0.1
0.1
0.1

0
0.2032
0.4064
0.6096
0.8128
1.016

0
0.032
0.068
0.099
0.124
0.18

0
0.032
0.068
0.099
0.124
0.18

0
0.032
0.068
0.099
0.124
0.18

0
0.0352
0.0748
0.1089
0.1364
0.198

0
0.0352
0.0748
0.1089
0.1364
0.198

0
0.0352
0.0748
0.1089
0.1364
0.198

0
0.0352
0.0748
0.1089
0.1364
0.198