Dear Rana,

Did you perform this analysis? I have to do exactly the same thing and I am wondering if you found any issues.

Thank you very much!

Dear Rana and C Filipe,

Did any of you done this analysis on the nanodrop already?

If so how did it work out and what did you do? Waiting for response..

Thank you!

Dear Filipe (sorry for the late reply) and Dear Alex:

I actually used this method and it worked very well for me. I can depend on the absorbance results to exclude the haemolysed samples from my cohort. however I noticed that samples with lipaemia give also a high absorbance at 414. and I am excluding those too.

Best luck.

Rana

Hi, thank you for your reply!

I have 1 remaining question, which method did you use from the nanodrop? Was it the UV/VIS protocol?

Because when I tried it, I wanted to measure the absorbance at wavelength's like 400, 380 and 415nm and it wasn't possible to choose them.. same as 380.. I only could choose to measure at for example 379nm and 382nm..

So which method did you use to measure hemolysis?

Thanks for helping

Dear Alex,

I also used this method. It worked great. Like Rana said, the lipaemic samples can also give strange spectrums and should be excluded too.

I used the UV-VIS function and it worked fine.. I was able do set the measures at 414nm and another one in each sample. I don't know how to solve your issue..

In a few weeks, I will have to work on this again. Perhaps we can talk about it then. Besides the identification of hemolytic samples, I also want to translate the given absorbance to free-hemoglobin value, and we can share our protocols.

Cheers, Carlos 

that's really a great idea Filipe thanks for sharing.

Dear Alex: we set our nanodrop as follows:

method type: manually entered factor/extinction cooefficient

measurement:

analysis wavelength:414 nm

result name: concentration

result unit: ng/ul

molecular weight 1user defined factor: 1 ng/ul

under correction:

use single point: 414 nm

baseline correction: no

under additional measurement:

Do post processing on measurement: No

under instrument setting:

display range: 414nm - 840 nm

is the pathlength automati.....: No

so you need to set up a new method for the haemolysis