meduser
07-27-2006, 10:56 PM
For those of you who don't know, a CHROMATOGRAPH is the data report that is produced by a HIGH PRESSURE LIQUID CHROMATOGRAPH, which is one of the MOST accurate ways of testing for THC level and for a CANNABINOID PROFILE of your medicine. (Gas Chromatography is slightly more detailed in some areas)
Here we go with Instructions on how to read them and what they show, along with chromatographs for 4 different mesh sizes of BUBBLEHASH (from one run) as well as the Chromo for some BHO OIL.
Enjoy!
*Note the 100 iu and 45 iu Bubblebags which produced hash that was over 50% THC!!
**The BHO sample comes out even higher at over 65% THC!! (and that is low in the BHO realm)
Cannabinoid Chromatography 101
Chromatography is often called separation science since it is a technique for separating compounds in a matrix. In the study presented here we are separating cannabinoids from a matrix of plant material, including chlorophyll, fats, protein and various other related compounds including the essential oils.
Cannabinoid chromatography focuses on five compounds: Delta 8 and Delta 9 Tetrahydrocannabinol (^8, 9-THC), Cannibinol (CBN), Cannabidiol (CBD) and Canabinolic Acid (THC-A). The most important being the THC-A in Liquid Phase Chromatography as is the practice here, as opposed to gas chromatography, which is practiced in 99% of other laboratories. The vital difference between gas and liquid phase chromatography is that in gas chromatography the sample is heated to greater than 200 degrees centigrade to perform the analysis in the gas phase. In heating the sample the THC-A is converted 1:1 to ^9-THC. Understanding this principle is key to making oral cannabis medicines.
Each of the five cannabinoids has a distinct chemical structure, that is, the carbon, hydrogen and oxygen atoms that make up the different cannabinoids differ in number and spatial arrangement giving them distinct chemical properties.
Two of these chemical properties are exploited by LC Chromatography, the first being their retention properties on a silica based, hydrocarbon coated column on which they are separated. Because of the slightly differing chemistries of the individual cannabinoids they will travel along this column at different rates carried by an organic solvent in which they are dissolved. They therefore enter the detector at different times from the beginning of the run. Subsequently, they enter a beam of UV light, which they absorb, again differently, depending on their chemical structures. This UV absorption is also characteristic of the particular compound and can be used for identification purposes.
In order to confirm which cannabinoid is which and how much of each one there is in our test solution we must run pure standards on the LC. These are the individual cannabinoids in pure form made up to a specific known concentration. These standard runs are used to calibrate the LC. That is, we can use these standards to tell the computer the concentration of each peak made by each compound. The peaks are made by the absorption of UV light by the compound and the area under the peak is directly proportional to the concentration of the test solution. Therefore, knowing the peak area of the standard allows us to calculate the concentration of a sample from its peak area.
In summary, by using pure standards of the individual cannabinoids we can quantify cannabinoids in samples and therefore determine their concentration. Furthermore, the pure standards allow us to identify the individual cannabinoids based on their retention times (time from injection to entering the detector) and their UV spectra.
In the chromatograms on the following pages a few points are in order:
The total THC potential refers to the percentage of THC which will become available on heating, for example smoking or baking into brownies. To calculate THC potential we have to add the % Amount values next to the names ^9-THC and THC-A. The total THC potential is then realized. Note that the ^9-THC in unburned samples is often less than 1% and that the THC-A can be as high as 25%*. CBN and CBD remain constant on heating. ^8-THC is often present in small amounts and also being psychoactive is included with the ^9-THC, both chromatographically and hence in calculations.
CBD/^9 ratios are provided on a number of chromatograms, this value is given for the ‘ground state’ cannabis, before it is heated. Obviously, this value will change dramatically after heating the sample, approaching infinity :). The ground state value is used to provide an indication of the CBD content, since this compound is an important modulator of the ^9-THC effect.
* We had one sample in our lab that measured out at 25% THC-A and 1% ^9-THC giving it a total THC potential of 26%. Samples with this level of THC potential are rare.
__________________________________________________ _____________
Here we go with Instructions on how to read them and what they show, along with chromatographs for 4 different mesh sizes of BUBBLEHASH (from one run) as well as the Chromo for some BHO OIL.
Enjoy!
*Note the 100 iu and 45 iu Bubblebags which produced hash that was over 50% THC!!
**The BHO sample comes out even higher at over 65% THC!! (and that is low in the BHO realm)
Cannabinoid Chromatography 101
Chromatography is often called separation science since it is a technique for separating compounds in a matrix. In the study presented here we are separating cannabinoids from a matrix of plant material, including chlorophyll, fats, protein and various other related compounds including the essential oils.
Cannabinoid chromatography focuses on five compounds: Delta 8 and Delta 9 Tetrahydrocannabinol (^8, 9-THC), Cannibinol (CBN), Cannabidiol (CBD) and Canabinolic Acid (THC-A). The most important being the THC-A in Liquid Phase Chromatography as is the practice here, as opposed to gas chromatography, which is practiced in 99% of other laboratories. The vital difference between gas and liquid phase chromatography is that in gas chromatography the sample is heated to greater than 200 degrees centigrade to perform the analysis in the gas phase. In heating the sample the THC-A is converted 1:1 to ^9-THC. Understanding this principle is key to making oral cannabis medicines.
Each of the five cannabinoids has a distinct chemical structure, that is, the carbon, hydrogen and oxygen atoms that make up the different cannabinoids differ in number and spatial arrangement giving them distinct chemical properties.
Two of these chemical properties are exploited by LC Chromatography, the first being their retention properties on a silica based, hydrocarbon coated column on which they are separated. Because of the slightly differing chemistries of the individual cannabinoids they will travel along this column at different rates carried by an organic solvent in which they are dissolved. They therefore enter the detector at different times from the beginning of the run. Subsequently, they enter a beam of UV light, which they absorb, again differently, depending on their chemical structures. This UV absorption is also characteristic of the particular compound and can be used for identification purposes.
In order to confirm which cannabinoid is which and how much of each one there is in our test solution we must run pure standards on the LC. These are the individual cannabinoids in pure form made up to a specific known concentration. These standard runs are used to calibrate the LC. That is, we can use these standards to tell the computer the concentration of each peak made by each compound. The peaks are made by the absorption of UV light by the compound and the area under the peak is directly proportional to the concentration of the test solution. Therefore, knowing the peak area of the standard allows us to calculate the concentration of a sample from its peak area.
In summary, by using pure standards of the individual cannabinoids we can quantify cannabinoids in samples and therefore determine their concentration. Furthermore, the pure standards allow us to identify the individual cannabinoids based on their retention times (time from injection to entering the detector) and their UV spectra.
In the chromatograms on the following pages a few points are in order:
The total THC potential refers to the percentage of THC which will become available on heating, for example smoking or baking into brownies. To calculate THC potential we have to add the % Amount values next to the names ^9-THC and THC-A. The total THC potential is then realized. Note that the ^9-THC in unburned samples is often less than 1% and that the THC-A can be as high as 25%*. CBN and CBD remain constant on heating. ^8-THC is often present in small amounts and also being psychoactive is included with the ^9-THC, both chromatographically and hence in calculations.
CBD/^9 ratios are provided on a number of chromatograms, this value is given for the ‘ground state’ cannabis, before it is heated. Obviously, this value will change dramatically after heating the sample, approaching infinity :). The ground state value is used to provide an indication of the CBD content, since this compound is an important modulator of the ^9-THC effect.
* We had one sample in our lab that measured out at 25% THC-A and 1% ^9-THC giving it a total THC potential of 26%. Samples with this level of THC potential are rare.
__________________________________________________ _____________