Testing CBD: HPLC Method

WRITTEN by Edd

 | Last revised

Have you ever wondered how companies test the cannabinoids that are in their hemp extracts?

When you buy CBD or any other cannabis product, it will have a labelled potency and total CBD content, along with other cannabinoids.

This information is determined by laboratory testing to find the concentration of major cannabinoids such as THC and CBD in the product.

HPLC (High-performance liquid chromatography) is one of the most widely used and reliable methods of testing cannabinoid standards.

If you have ever looked at a CBD product’s Certificate of Analysis, you have probably looked at results created using this analysis method.

So, knowing the basics of how HPLC works will help you understand those lab results and make more informed decisions about buying CBD and other cannabis products.

What Is HPLC Analysis Of Cannabinoids?

Chromatography is a method of identifying the separate components of a solution using the differing rates at which each component travels through a substrate.

It was originally created to separate pigments used in dyes so that clothing makers could know the precise amounts of each pigment used to create a dye.

Early chromatography simply separated the various components to be studied with the naked eye.

Modern methods involve the precise detection of UV light, spectroscopy, and other methods, to analyse samples more accurately than just by looking for changes in colour with the naked eye.

High-Performance Liquid Chromatography, or HPLC, is a method of chromatography that uses high pressure to push the liquid sample, suspended in another solution called the mobile phase, through a column of porous material to a second solution. [1]

The difference in solubility between the sample and the second solution causes the different compounds in the sample to move through the column at different speeds.

This allows for the detection of the quantity of each component individually, using various types of detectors.

HPLC is used to quantify the precise components of many different products intended for consumption or other uses. For example, histamine HPLC methods are used to determine the freshness of seafood, as histamine levels increase over time in dead fish tissue.

Man in a laboratory looking at CBD oil

Cannabinoid Analysis HPLC

Because of the strict regulations in place controlling the contents of cannabis products, accurate analysis of their components is an important part of buying, selling, and producing high-quality cannabis products.

HPLC is the most common method of analysing products that contain cannabinoids, along with gas chromatography. Both of these methods are used in the creation of a cannabis product’s Certificate of Analysis.

Knowing how HPLC separation of cannabinoids works will help you to understand how the contents of a cannabis product are measured and the limitations of this method that could affect the accuracy of the lab test results and potencies you see on cannabis product labels.

HPLC For Cannabinoids

The parameters of an HPLC cannabinoids test can differ significantly depending on which compounds the analysis is looking for. This includes a wide range of factors, such as: [2]

  • The temperature of the liquid sample
  • The concentration of the liquid sample
  • The type of porous material used in the columns
  • The amount of pressure used in the test

In addition, different mobile phases for HPLC with cannabis are needed to detect different cannabinoids. Furthermore, the HPLC flow rate cannabinoids need also differs for each cannabinoid.

Because of these factors, each HPLC method can only detect a small number of targeted compounds.

Cannabis is known to contain more than 100 different cannabinoids. As a result, no single test can detect every cannabinoid, though most tests cover all the important ones.

Some CBD manufacturers will have their products tested by several different laboratories in order to cover a wider range of testing methods. [3]

Product testing laboratories usually rely on HPLC systems that can identify and quantify the most common cannabinoids found in cannabis, such as CBD and THC.

In other words, even the most thoroughly tested products can contain traces of less common cannabinoids that do not show up in test results for any of the HPLC methods used.

Man in a laboratory holding CBD oil and hemp flower

Cannabinoid Standards For HPLC

There is a range of HPLC methods used by various independent testing companies to assess the contents of a cannabis product.

Testing methods are not standardised between different testing laboratories yet, even within one country. Testing standards can differ even more when looking at test results from another country.

Some of the more common methods include:

Agilent Cannabinoids HPLC Analysis

Agilent is a company that produces scientific equipment for laboratories. The company has published methods of testing for cannabinoids for its HPLC systems that can detect 11 of the most common cannabinoids. [4]

Confidence Analysis HPLC UV

Confidence Analysis is a testing laboratory that analyses cannabis and hemp products. Its method of assessing CBD potency can detect eight cannabinoids by HPLC UV. HPLC UV uses an ultraviolet light detector to identify each component in a sample. [5]

Alliance HPLC

Alliance employs one of the most comprehensive cannabinoid HPLC systems, which is capable of quantifying 16 different cannabinoids. However, this only covers a fairly small number of the 140+ cannabinoids found in the cannabis plant. [6]

HPLC Is A Precise Analysis Method For Cannabinoids

HPLC can accurately and precisely measure the different compounds in your cannabis products.

While it cannot test for every cannabinoid, most HPLC testing methods will cover all the major cannabinoids found in the extract.

However, most of the commonly used methods will identify the most common cannabinoids and any unsafe contaminants.

So even though you cannot use HPLC analysis to fully understand a cannabis product’s effects, it does provide an accurate picture of a product’s quality and consistency.

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References

1. Vera Koester, "What Is HPLC?".
ChemistryViews.
https://www.chemistryviews.org/details/education/9464911/What_is_HPLC/.
2. Masoumeh Pourseyed Lazarjani et al., "Methods for quantification of cannabinoids: a narrative review".
Journal of Cannabis Research.
https://jcannabisresearch.biomedcentral.com/articles/10.1186/s42238-020-00040-2.
3. N/A, "Cannabis (Marijuana) and Cannabinoids: What You Need To Know ".
National Center for Complementary and Integrative Healt.
https://www.nccih.nih.gov/health/cannabis-marijuana-and-cannabinoids-what-you-need-to-know.
4. Christy Storm, Michael Zumwalt, and Anthony Macherone, "Dedicated Cannabinoid Potency Testing in Cannabis or Hemp Products Using the Agilent 1220 Infinity II LC System ".
Agilent Technologies.
https://www.agilent.com/cs/library/applications/application-dedicated-cannabinoid-potency-testing-5991-9285-en-us-agilent.pdf.
5. Pat Reynolds, "Eight Cannabinoids by HPLC-UV ".
Confidence Analytics.
https://www.conflabs.com/eight-cannabinoids-by-hplc-uv/?doing_wp_cron=1655745567.8364210128784179687500.
6. Andrew J Aubin, Catharine Layton, and Shawn Helmueller, "Separation of 16 Cannabinoids in Cannabis Flower and Extracts Using a Reversed Phase Isocratic HPLC Method".
Waters.
https://www.waters.com/waters/library.htm?cid=534293&lid=135003295.

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Disclaimer. While we strive to relay the most factual education available, this shouldn’t replace official medical or legal consultation and recommendation. This is for educational and entertainment purposes only. Happy days.

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