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# Kd Calculation Question

## Kd Calculation Question - Chromatography Forum

### Kd Calculation Question - Discuss Chromatography techniques in this forum. Includes a HPLC forum, and other "liquid phase" separation techniques.

#1
04-16-2009, 08:47 PM
Pipette Filler

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Kd Calculation Question

hello
i will explain to you what i do with some data for example to learn me how i can do this with any software you can tell and send me link for it.
we make quality control for product as followed:
we take the product and dissolve it in injection buffer (1ml) which consist of elution buffer and contain sodium azide then apply this to column chromatography then receive the eluted in test tube then measure this with spectrophotometer , from this data we determine Kd for the polysacchride in product as the following equation
kd= (Ve-Vo)/(Vt-Vo)
Ve is elution volume for the polysacchirde
Vo is elution volume for polydextran (calibration standard)
Vt is elution volume for sodium azide
this Kd must be equal to 0,4 or less
then we plot optical density on Y axis against volume on X axis to calculate
recovery percentage assuming that Kd equal or smaller than 0,5 at this equation
(area in evaluated peak of polysacchride at Kd equal 0,5/total area in evaluated peak)
and the result multiply it with 100 to get percentage
this all work

2 = 0.447
4= 0.391
6 =0.38
8= 0.327
10= 0.387
12 =0.366
14 =0.376
16 =0.33
18 =0.385
20 =0.361
22 =0.378
24 =0.347
26= 0.527
28 =0.672
30 =0.732
32= 0.436
34 =0.448
36= 0.399
38 =0.465
40 =0.428
42 =0.444
44 =0.428
46 =0.461
48 =0.451
50 =0.453
52 =0.443
54 =0.464
56 =0.422
58 =0.449
60 =0.405
62 =0.426
64 =0.373
66 =0.396
68 =0.358
70 =0.387
72 =0.329
74 =0.364
76 =0.333
78 =0.372
80 =0.338
82 =0.39
84 =0.356
86 =0.371
88 =0.341
90 =0.395
92 =0.471
94 =0.438
96 =0.519
98 =0.469
100 =0.573
102 =0.596
104 =0.408
106 =0.475
108 =0.36
110 =0.397
112 =0.342
114 =0.377
116 =0.335
118 =0.411
120 =0.344
please this is the data help me to how i can calculate with any software and send this link for this software

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