# dilution factor

Hello all,

I need some help understanding the agilent application note: Quantification of THC and CBD in Gummies and Hard Candies, specifically where the dilution factor is calculated

The SOP states:

1g gummy into 10ml water, then homogenise

add 10ml ACN to solution, homogenise

add quechers salt, shake + centrifuge

top 10ml ACN layer contains analytes

remove 2ml of the supernatant.

filter into sample vial

when i use x10 DF factor i am getting really high numbers which dont add up. Can anyone explain where the DF x10 comes from? I emailed one of paper authors and he just repeats that its x10.  his reply was "You have a good question. The dilution is indeed 10X because you finish in 10 ml acetonitrile. In other words, you go from 20ml to 10 ml."  but surely going from 20ml to 10ml, is a dilution factor of 0.5

im really confused, please help. i typicall use final-volume/initial-volume (or aliquot) to determine DF which seems to work, except in this example

Tan

Parents
• Step 3 adds the first 10 mL ACN/NH3 - 1 g in 10 mL
Step 5 adds another 10 mL ACN/NH3 - 1 g in 20 mL

Step 10 you take 2 mL out of 20 mL, therefore the DL is 10 = 2/20

• the paper i look at STEP 3 is just 10ml water. Step 5 ACN/NK3

i believe the quechers salts separates the water (10ml) from the ACN/NH3 (10ml)

• True, you will have a ca. 10 mL supernatant layer of ACN, where you take 2 mL from, which is a 1:5 ratio. Just ignore the DL in the text, it changes with adding standards or using different volumes of solvents. Again, be sure about the absolute amounts of the sample and what you analyze for your calculations. I will contact the author to remove the sentence from the application note.

• Thanks.  That being said, in my opinion the sample NEEDS dilution, maybe not x10 but def some.

I have THC and CBN calibrated from 1ug/ml -200ug/ml and when i perform the prep as stated my results is way beyond the calibration curve. I was going to start diluting this.

yes, thanks I will be sure to take care to make sure the amounts are correct. thanks for the help

• Thanks.  That being said, in my opinion the sample NEEDS dilution, maybe not x10 but def some.

I have THC and CBN calibrated from 1ug/ml -200ug/ml and when i perform the prep as stated my results is way beyond the calibration curve. I was going to start diluting this.

yes, thanks I will be sure to take care to make sure the amounts are correct. thanks for the help

Children
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