Federal researchers at the National Institute of Standards and Technology (NIST) say they’ve made the first-ever detection of THC in human breath following the consumption of infused edibles—a possible step forward in terms of field testing for cannabis impairment.

But more research is necessary, they add, pointing to “uncertainty of breath measurements” and the need to further analyze the detectability of cannabinoids over longer time periods.

“This is an important step forward, that we can detect THC increases in breath after the ingestion of cannabis,” Jennifer Berry, a NIST research chemist and the lead author on the paper, said in an agency release about the new findings, which were published earlier this month in the Journal of Analytical Toxicology.

Three NIST researchers and one from University of Colorado Anschutz Medical teamed up on the project, which the report describes as a “proof-of-concept study.” It’s part of the federal agency’s broader research into cannabis use and driving performance.

Breath samples of 29 participants were collected before and after ingesting a THC-infused edible, which subjects were asked to bring in themselves. The products ranged in strength from five milligrams to 100 mg THC, though the report notes the amounts were not independently verified.

“NIST and its partners at University of Colorado Anschutz have made the first measurement of tetrahydrocannabinol (THC) in breath from edible cannabis.”

Collection was done using two different devices, one described as an aerosol device and the other as a condensate device. Samples were taken before ingestion and at three timepoints afterward: roughly 47 minutes, 92 minutes and 180 minutes.

Though participants were instructed to abstain from using edible cannabis for 12 hours prior to the study and from inhaling cannabis for 8 hours ahead of time, samples for nearly all participants detected THC even before they ingested the infused edibles.

“THC was detected in breath samples from 27 of the 29 participants before edible ingestion, ranging from trace (10 participants) to greater than 0.4 ng/device,” the report says, noting that similar phenomena around residual THC had been observed in prior studies.

“The finding of detectable and/or quantifiable THC before cannabis use,” authors wrote, “shows how challenging it is to interpret a single THC concentration measurement in breath with no prior information about THC at baseline.”

“The high detection rate of THC in breath after 8 h of requested abstinence,” they added later in the report, “shows that identifying recent use from a single measurement is challenging from any matrix.”

After ingesting the edibles, measured THC levels in the breath of most participants—but not all—rose over time.

“Of the 29 participants in this study, 19 showed a significant increase in THC concentration after edible ingestion (at any of the three post-use timepoints), 4 showed no change, and 6 showed a significant decrease in THC concentration after edible use,” the study says.

Those who showed a decrease in THC levels, it points out, also reported some of the highest THC levels prior to taking the edibles.

Those whose THC breath levels rose or remained stable, however, support the notion that breath-based tests can detect recent cannabis use, authors asserted.

“Breath samples from two-thirds of the participants in this study showed an increase in THC concentration after cannabis ingestion,” they wrote, “but the remaining one-third are not necessarily inconsistent with the hypothesis that recent cannabis use can be detected in breath, regardless of the mode of use.”

Notably, measured levels of THC and some other cannabinoids seem to change in similar ways. But THC and CBD levels didn’t always track with each other.

“When detected, CBN and CBG matched THC trends, but there were multiple instances where CBD and THC disagreed,” the study says, “suggesting differences in biological processing or clearance.”

In the NIST press release, Tara Lovestead, a chemical engineer on the breath research project, said the study “supports the idea that multiple breath measurements over a period of time could be a way to use a breathalyzer to detect cannabis use, regardless of how it’s ingested.”

“However,” the scientist added, “devices will still need standards to ensure that they are accurate and used correctly, standards that don’t yet exist.”

NIST notes in its press release that the agency itself is not developing a cannabis breath-testing device. “Instead, NIST’s role is to help ensure that measurements of cannabis in breath can be accurate, reliable and have scientifically sound standards behind them,” it says.

“Looking forward, we can now tackle the question of when THC increases after edible ingestion, when it goes back to baseline, and how to analyze breathalyzer data to get the information needed.” said Kavita Jeerage, a NIST research chemical engineer who the agency’s cannabis breath research.

The agency is set to host a workshop with device makers in September.

The event was originally scheduled to be held in Boulder, Colorado, in April, but a month ahead of the originally scheduled event, it was abruptly postponed.

According to a preliminary agenda sent to Marijuana Moment by NIST earlier this year, topics to be discussed will include challenges facing marijuana breathalyzer design and development, obstacles to prosecutors handling drugged-driving cases and how NIST and others might partner to advance the technology.

Findings of the workshop, “Building a Path Forward for Meaningful Cannabis Breathalyzer Realization,” are set to be compiled into a NIST internal report that the agency has said will be publicly available.

Unlike with alcohol, there’s currently no widely accepted field test to determine whether someone is under the influence of marijuana.

In 2023, a federally funded report by researchers at NIST and the University of Colorado Boulder concluded that evidence does “not support the idea that detecting THC in breath as a single measurement could reliably indicate recent cannabis use.”

“A lot more research is needed to show that a cannabis breathalyzer can produce useful results,” Jeerage at NIST said at the time. “A breathalyzer test can have a huge impact on a person’s life, so people should have confidence that the results are accurate.”

More recently, a U.S. Department of Justice researcher cast doubt on whether a person’s THC levels are even a reliable indicator of impairment.

States may need to “get away from that idea,” Frances Scott, a physical scientist at the National Institute of Justice (NIJ) Office of Investigative and Forensic Sciences under DOJ, said on a podcast early last year.

Scott questioned the efficacy of setting “per se” THC limits for driving that some states have enacted, making it so a person can be charged with driving while impaired based on the concentration of cannabis components in their system. Ultimately, there may not be a way to assess impairment from THC like we do for alcohol, she said.

One complication is that “if you have chronic users versus infrequent users, they have very different concentrations correlated to different effects,” Scott said. “So the same effect level, if you will, will be correlated with a very different concentration of THC in the blood of a chronic user versus an infrequent user.”

That issue was also examined in a federally funded study last year that identified two different methods of more accurately testing for recent THC use that accounts for the fact that metabolites of the cannabinoid can stay present in a person’s system for weeks or months after consumption.

The THC impairment question has nevertheless been a major focus for lawmakers and the research community, particularly as it concerns driving laws.

Last October, a study preprint posted on The Lancet by an eight-author team representing Canada’s Centre for Addiction and Mental Health, Health Canada and Thomas Jefferson University in Philadelphia identified and assessed a dozen peer-reviewed studies measuring “the strength of the linear relationship between driving outcomes and blood THC” published through September 2023.

“The consensus is that there is no linear relationship of blood THC to driving,” the paper concluded. “This is surprising given that blood THC is used to detect cannabis-impaired driving.”

Most states where cannabis is legal measure THC intoxication by whether or not someone’s blood THC levels are below a certain cutoff. The study’s findings suggest that relying on blood levels alone may not accurately reflect whether someone’s driving is impaired.

“Of the 12 papers included in the present review,” authors wrote, “ten found no correlation between blood THC and any measure of driving, including [standard deviation of lateral position (SDLP)], speed, car following, reaction time, or overall driving performance. The two papers that did find a significant association were from the same study and found significant relationship with blood THC and SDLP, speed and following distance.”

Earlier last year, researchers behind a federally funded study said they’d developed new procedures to enhance the selectivity of a popular forensic testing method, allowing better detection of delta-9 THC and its metabolites in blood.

A 2023 congressional report for a Transportation, Housing and Urban Development, and Related Agencies (THUD) bill said that the House Appropriations Committee “continues to support the development of an objective standard to measure marijuana impairment and a related field sobriety test to ensure highway safety.”

A year earlier Sen. John Hickenlooper (D) of Colorado sent a letter to the Department of Transportation (DOT) seeking an update on that status of a federal report into research barriers that are inhibiting the development of a standardized test for marijuana impairment on the roads. The department was required to complete the report under a large-scale infrastructure bill that President Joe Biden signed, but it missed its reporting deadline.

A study published in 2019 concluded that those who drive at the legal THC limit—which is typically between two to five nanograms of THC per milliliter of blood—were not statistically more likely to be involved in an accident compared to people who haven’t used marijuana.

Separately, the Congressional Research Service in 2019 determined that while “marijuana consumption can affect a person’s response times and motor performance … studies of the impact of marijuana consumption on a driver’s risk of being involved in a crash have produced conflicting results, with some studies finding little or no increased risk of a crash from marijuana usage.”

Another study from 2022 found that smoking CBD-rich marijuana had “no significant impact” on driving ability, despite the fact that all study participants exceeded the per se limit for THC in their blood.

Evan as far back as 2015, a U.S. National Highway Traffic Safety Administration (NHTSA) concluded that it’s “difficult to establish a relationship between a person’s THC blood or plasma concentration and performance impairing effects,” adding that “it is inadvisable to try and predict effects based on blood THC concentrations alone.”

In a separate report last year, NHTSA said there’s “relatively little research” backing the idea that THC concentration in the blood can be used to determine impairment, again calling into question laws in several states that set “per se” limits for cannabinoid metabolites.

“Several states have determined legal per se definitions of cannabis impairment, but relatively little research supports their relationship to crash risk,” that report said. “Unlike the research consensus that establishes a clear correlation between [blood alcohol content] and crash risk, drug concentration in blood does not correlate to driving impairment.”

Earlier this month, meanwhile, a Rutgers University law professor published commentary arguing that roadside tests for marijuana impairment used widely by police departments across the country are “inadequate” and “pseuduscientific,” urging policymakers to take a more evidence-based robust approach to transportation safety while relying less on law enforcement’s ostensible expertise.

The paper, by William J. McNichol of Rutgers University’s Camden School of Law, asserts that current approaches to detecting cannabis impairment “either merely imitate tools used to identify alcohol based impairment without taking into account important differences between the effects of these molecules or they are pseudoscientific ‘police science.’”

Original publication: November 1986.

BREEDING FOR BIG BUDS

HARVEST ’86

BY JORGE CERVANTES

“Look at the size of that thing, I must be hallucinating,” said Tim as he stared in awe at the huge six-inch diameter flower top. “What did you put in this joint to make me see things twice as big as life? I mean, that is the biggest bud I’ve ever seen!”

“We call her Big Bud. I think she is 100% Indica,” replied James with a big smile. “I brought her back to Detroit from out West. Here, give her a try,” said James, passing Tim a bomber of Big Bud.

Tim put the spliff between his lips and gently drew a few breaths through the unlit joint. “So fresh and minty, it’s the best I’ve ever tasted. I take back all the bad stuff I said about indoor smoke,” Tim said with a pleased sheepish grin.

“They’ve been breeding some incredible plants out on the West Coast. When I was there, indoor growers were talking about the Hash Plant that matures into a 36″ tall two-three ounce plant (dried tops only) after eight weeks of flowering: stout, broad tops laden with thick gooey resin. Or Lime Bud, a Sativa/lndica cross, with long, dainty, lime green leaves and lightweight buds packed with pistils and glistening resin glands. Or Northern Lights, a mainstay variety in Vancouver, B.C. that matures into a four-foot, four ounce plant at the ripe old age of 85 to 90 days. But everyone’s favorite is Big Bud because she grows huge buds and has the sweetest, mintiest taste in the world. My last crop weighed in at 3 1/2 pounds, that’s better than four ounces a plant!”

Narrow leaves show classic qualities.

All of these varieties were developed by industrious, careful and caring individuals that have kept meticulous detailed records of plant growth habits and the offspring that resulted when they crossed a male plant with a female. Indoor breeding is two or three times faster than outdoors. As many as three generations may easily be completed in one year indoors. The breeder controls the hours of light and darkness per day, causing the plants to flower and produce seed at will.

Selective breeding (also known as selective sexual propagation) has completely changed the quality and yield of indoor marijuana crops. The starting point is to collect marijuana seed from well known pot growing regions throughout the globe. The various seed strains are grown and studied thoroughly without any cross breeding. Once each is understood, then selective breeding can begin, either to attain a pure seed stock, or to improve the strain. Once the ultimate plant is developed through this sexual propagation, it can be cloned to preserve the genetic blueprint created by selective breeding.

Cannabis Indica seeds are highly prized for their potency and adaptability to the indoor environment. The characteristics of Indica plants include: squat, bushy, vigorous growth; early, sustained potency; disease resistance; and heavy bud yield. An Indica plant may yield two to four times the dry bud weight of a Sativa at harvest. Thai, Colombian, and Mexican are examples of Cannabis Sativa. These Sativa varieties are tall, lanky, and disease resistant, with narrow, long leaves and light buds that take up to three months to mature. But the high is energetic, soaring, and clear.

Ideally, skilled breeders will strive to retain the desirable dominant growth characteristics of both Cannabis Sativa and Cannabis Indica to form super strains. Breeders may cross a Sativa with an Indica, hoping to retain the desirable soaring high while keeping the short, stocky growth characteristics and early maturity of the Indica.

This single indica female bud is being fertilized by male pollen in the plastic baggie.

Selective breeding requires the breeder to simply assume the role of Mother Nature once again. In nature, pollen from the male cannabis plant is shed into the wind to randomly fertilize any receptive female plant. A breeder adds precision and control to this process by catching pollen from a desirable male and carefully placing it in contact with chosen female pistils.

Two basic kinds of breeding are: (1) inbred or true bred—plants of the same strain or ancestry that are crossed with one another. (2) outbred or hybrid— plants of different strains that are crossed or cross-pollinated.

The goal of inbreeding is to establish a pure breed to start from. Selected females are pollinated with male pollen of the same strain, then the resulting seeds are planted and checked for variability. This process may take several generations to produce a strain that actually breeds true. Pure bred plants are grown until the fifth to sixth generation, after which negative characteristics, like low potency, legginess and lack of vigor tend to dominate.

This true or pure breed is essential to establish common growth characteristics. Without a pure breed on one or both sides, it is difficult to predict the outcome of a hybrid cross. Ideally, both male and female are pure-bred plants, with known ancestry and growth attributes.

A hermaphroditic plant (a plant displaying both male and female flowers) will self-pollinate and produce inbred seeds. Although this is a shortcut to inbreeding, the seedlings resulting from self-pollination may tend to be hermaphroditic themselves. This can be a real headache for the sinsemilla grower or breeder, with unwanted seeds and uncontrolled pollen. Nonetheless, the seed produced by a hermaphrodite that has just a few male flowers on a female plant will be 80-90% female seed. One easy way to produce seeds from an all female clone crop is to stress one of the female branches by twisting a piece of wire around one of the branches a couple of weeks before flowering is induced. The extra stress will usually cause the plant to produce male flowers.

Outbreeding, or producing hybrid seed, has increased agricultural production more than anything. Farmers, breeders and cannabis horticulturists have found that crossing parents of different pure strains, especially those exhibiting exceptional, dominant, complementary characteristics, will result in a super plant, also referred to as hybrid vigor.

It is possible to cross Sativa/lndica. Some growers swear by the cross, saying you get the best of both plants: vigor and size from the Sativa, and squat, bushy, early, potent growth from the Indica.

Choosing from a large and varied plant stock is the key to successful breeding. Since the origin of most seed is dubious, an accomplished breeder waits to see what kind of plant it produces, before deciding which plants to cross to form hybrid seed. If the breeder plants only a few seeds, there is no guarantee that they will grow into ideal breeding stock, even if they came from dynamite smoke. If the breeder instead selects the best plants from many seedlings of various strains of Cannabis Indica (and/or Sativa), he or she will have a better chance of achieving the desired outcome.

BREEDING: STEP-BY-STEP

STEP ONE: Choose male breeding stock exhibiting desirable characteristics.

STEP TWO: One branch full of male flowers is all that will be needed, unless a large crop is desired. Unused branches may be stripped of flowers to help contain pollen and guard against accidental, random pollination. The male can be isolated from the females, once flowers have developed, but not yet opened, by placing him in a sunny window or in a vegetative grow room. This will slow flower development and not hurt the male in any way.

A branch of ripe, male flower may be cut and placed in water. It will remain healthy for several weeks. When the pollen sacks open, proceed with Step Three. The remaining male plant may then be cut back or harvested.

Skilled breeders will cross a Sativa with an Indica, hoping to retain the Sativa’s soaring high while keeping the stocky growth and early maturity of the Indica.

STEP THREE: When the pollen pods start to open, place a clean, plastic sack or baggie over the branch to collect pollen. Secure the bag at the bottom with a piece of string or wire tie. Keep the bag over the branch for several days to collect pollen.

STEP FOUR: When enough pollen has been collected, shake remaining pollen off into the bag. Remove spent branch. Pollen may be kept for several weeks by removing all vegetative matter to prevent mold, and storing the pollen in a dark, dry container in the refrigerator.

STEP FIVE: Ideally, pistils should be ready for fertilization three to four weeks after the first calyx has appeared. Receptive pistils are white and fuzzy, not starting to turn brown. Cover the selected female branch that has many ripe, receptive pistils with the pollen-filled bag. Shake the bag.

STEP SIX: Use a small paint brush to apply the pollen from the bag to the pistils if just a few seeds are desired from many different females. Be very careful. Just use a little pollen on each calyx and keep it from spreading to the sinsemilla crop.

STEP SEVEN: Leave the bag for two or three days, to ensure fertilization. Be careful not to scatter pollen on adjacent sinsemilla crop when removing the bag.

STEP EIGHT: After fertilization, seeds will be ripe in three-six weeks. Harvest seeds when they split open the containing calyx or rattle in the pod.

STEP NINE: Let seeds dry for two to three months, in a cool, dry place, before planting.

This is the eighth in a series of articles by Jorge Cervantes, author of the bestselling INDOOR MARIJUANA HORTICULTURE, known by many as the Bible of indoor growing. The series outlines indoor growing from A-Z, for both novices and experienced gardeners alike.