Excerpts from "Nutraceutical potential, phytochemistry of hemp seed and its application in food and feed: A review"

Excerpts from “Nutraceutical potential, phytochemistry of hemp seed (Cannabis sativa L.) and its application in food and feed: A review:”

“Studies suggest that these compounds [in hempseed] exhibit therapeutic potential against conditions such as epilepsy, certain neurological disorders (Parkinson’s and Alzheimer’s diseases), and cardiovascular issues. Additionally, hemp seeds possess other notable properties, such as anti-cancer, anti-inflammatory, and immunomodulatory effects. They are also used as a dietary supplement to alleviate headaches, insomnia, high cholesterol levels, and premenstrual syndrome symptoms in women.”

Schultz et al. (2020) used chemical analysis and immunolabeling for the study of complex carbohydrates present in hemp seed. They found that hemp contains crystalline cellulose as a major polysaccharide. Monosaccharide analysis and immunolocalization analysis showed xylan (non-cellulosic polysaccharide) as the second most abundant polysaccharide. Xylan acts a prebiotic in the gut, promoting good bacteria and thereby improving health. Besides there are certain antinutrients present in hemp seed like phytate, lignin, trypsin inhibitor and tannins with phytic acid most prevalent (22.5 mg/g) (Schlemmer et al., 2009). Mostly these antinutrients are located in the cotyledons of the seed. The anti-nutrient nature of phytate is because it binds or chelates with iron and zinc and reduce their bioavailability. It also affects the digestibility of proteins (Muzquiz et al., 2012; Zouaoui, Létourneau-Montminy & Guay, 2018). However, both lignin and phytate show their beneficial nature by acting as an antioxidant.”

Alkaloids
Alkaloids under most physiological conditions can act as antioxidants and hence protect plants against oxidative stress. It has been seen in humans that a statistically significant correlation between nutritional patterns of phenolic compounds and lower incidence of major illnesses such as cardiovascular diseases, neurological disorders and carcinoma (Arts & Hollman, 2005). However, the phenolic antioxidant properties might not always clearly describe these positive health benefits, as they are inadequately bioavailable. Hemp seeds and hemp sprouts of C. sativa are one of the richest sources in phytochemical compounds, particularly for these polyphenols viz., alkaloids such as caffeoyltyramine, cannabisin A, B, C and also saccharides and amino acids (Frassinetti et al., 2018).”

Flavonoids
Flavonoids are a group of compounds that includes polycyclic, aromatic structure with oxygen, and sometimes glycoside (sugar) linkages. Anthocyanins, natural pigment molecules, are a subgroup of flavonoids that play a major part in the phenotypical expressions & signature for purple and green hues. Flavonoids are regarded as an integral part in a various form of nutraceutical, medicinal and pharma-based applications, due to primarily their antioxidant, analgesic, anti-cancer & anti-teratogenic properties in a combination of its ability in altering fundamental enzymatic functions (Feng, Hao & Li, 2017). Isolation and structure identification of flavonoids has reported flavonoids, cannflavin A and B are two specific prenylated flavones with antioxidant activities (Panche, Diwan & Chandra, 2016).
Lately, there has been significant research attention on the connection between gut microbiota and the promotion of health (Soderborg et al., 2018; Wan et al., 2019). It is now widely recognized that gut microbiota play a role in influencing health status (Agus, Planchais & Sokol, 2018; Fan & Pedersen, 2021). The main actors in the transformation of flavonoids, particularly in the O-deglycosylation process, are the majority of gut microbiota, including Bifidobacteriaceae, Lactobacillaceae, Lachnospiraceae, and Enterococcaceae (Kim et al., 2020; Yan & Zhang, 2022). Typically, gut microbiota engage in the catabolism of flavonoids, and the process likely involves the regulation of gut microbiota. However, further exploration is needed to understand the regulatory mechanisms for specific flavonoids and extracts (Li et al., 2023).

Pharmacological properties of hemp seed

Antimicrobial
From long ago, hemp seed has been used as medicinal plant source and have been used in the treatment of some of the diseases like arthritis, asthma, menstrual cramps, glaucoma, atopic dermatitis, autoimmune diseases, cancer, hypertension and inflammatory diseases (Jeong et al., 2014; Jin et al., 2018; Leonard et al., 2020). There have been several studies which prove the antimicrobial property of hemp.

Nissen et al. (2010) hypothesized that the seed sowing time and plant age of industrial hemp can greatly affect the microbial inhibitory action. Furthermore, the essential oils derived from the inflorescences of industrial hemp, primarily the Futura variety, demonstrated significant antimicrobial efficacy against Gram-positive opportunistic and pathogenic bacteria.

Ali et al. (2012) studied the antibacterial effect of seed oil against standard fungi and bacteria and compared the antibacterial activity with reference drugs. The oil extracted from hemp seeds displayed notable antibacterial effects against Bacillus subtilis, akin to the potency of 20 µg/ml Gentamicin. It also exhibited inhibitory properties against Staphylococcus aureus, mirroring the effects of 10 µg/ml Benzyl penicillin and 40 µg/ml Cloxacillin. Additionally, it suppressed the growth of Escherichia coli in a manner comparable to 10 µg/ml Gentamicin and showed similar inhibitory action against Pseudomonas aeruginosa, as seen with 20 µg/ml Gentamicin.

A more recent study by Frassinetti et al. (2020), illustrated the antimicrobial effect against pathogenic strain and biofilm formation against S. aureus. The impact of hemp seed extract on the biofilm-producing strain S. aureus ATCC 35556 was assessed by measuring the percentage of viable versus deceased bacterial cells, utilizing a FD/PI double-staining assay. In the context of Gram-positive bacteria, there was an observed increase (approximately 15 %, p < 0.05 vs CNT mortality) in stained cells that were no longer alive. This increase was not as substantial as the effect caused by the highest concentration of hemp seed extract. Panel B presents the ratio of live to dead cells, indicating a mortality rate surpassing 50 % for all treatments, with the most prominent outcome (∼78 %) occurring after exposing the strain to 1 mg/ml of hemp seed extract. These findings strongly indicate a potent antibacterial influence of the 1 mg/ml hemp seed extract on S. aureus biofilm producers, potentially attributed, at least in part, to mechanisms that damage the bacterial cell membrane.

Antioxidant
It is reported that hydrophobic, acidic and branched-chain amino acids predominantly contribute to escalating antioxidant prospective of hemp seed protein hydrolysate (Girgih et al., 2014). Lu et al. (2010) performed peptide sequencing to analyse amino acids responsible for free radical scavenging. They used a specific technique like microporous adsorption resins (MARs) for separating bioactive constituent from the crude hemp seed protein isolate (HPI). It was observed that the peptide sequence had more than 50 % hydrophobic amino acid residue on both sides of the chain which favoured inhibition of oxidation. It also proved their theory to use an increased dose of HPI extract on PC12 cells to check cell viability. It was found that with increasing dosage there was a significant increase in cell survival.

A recent study by Logarušić et al. (2019) showed the effect of hemp seed protein hydrolysate (HPH) on normal HaCaT and cancer (HeLa) cell line, they observed 25.3 % inhibition of Reactive oxygen species (ROS) formation in pre-treated HPHsingle bondN and a 28.1 % in HeLa cell line. They also observed a stimulatory effect on normal cell growth and pointed out the possible use as a media supplement. Studies also indicate that hemp seed protein extraction method can affect optimum antioxidant activity (Girgih, Udenigwe & Aluko, 2013; Rezvankhah et al., 2019).

In addition, various tocopherol types, such as alpha, beta, gamma, and delta tocopherols, are present in hemp seeds in small amounts, around 0.1 %. These tocopherols belong to the vitamin E group and have antioxidant properties, which are important for human health and nutrition. Similarly, phenols and polyphenols in hemp seeds have antioxidant activity (Martinez et al., 2023)

Antifungal
Currently, there are numerous challenges to combat infections which are particularly caused by multidrug-resistant organisms. A study reports that α-humulene, a bioactive compound found in cannabis oil has proven effective potency against Cryptococcus neoformans. It causes lung infection and secondary infections like fungal meningitis and encephalitis infection in AIDS patient (Wanas et al., 2016). Another study found 9 new cannabinoids, among them 8-hydroxycannabinol was identified using 1D, 2D NMR spectroscopy and also showed antifungal property against Candida albicans (Radwan et al., 2009).

Antiinflammatory
Hemp seed contains PUFAs omega-3 and omega-6, which have shown the anti-inflammatory properties (Latif & Anwar, 2009; Leizer et al., 2000; Seo et al., 2012). Hemp seed (of Bama variety) is being used to reduce the spleen inflammation and systemic inflammation in female rats (Leonard et al., 2020). Additionally, Arab physicians have also been known to use the hemp because of its anti-inflammatory, antipyretic, painkilling, anti-epileptic, anti-emetic and diuretic properties (Fathordoobady et al., 2019; Lozano, 2001).

THC is one of the phytocannabinoids present in the hemp seeds which have 20 times more anti-inflammatory property than aspirin and double as that of hydrocortisone. In addition to tetrahydrocannabinol, hemp seed also contains CBD which has similar properties like anti-inflammatory and analgesic (Baron, 2018; EFSA, 2011; Fathordoobady et al., 2019). A large number of cannabidiol modulators containing amide are beneficial for the therapeutics of inflammation, rheumatoid arthritis, pain, allergies, nephritis, multiple sclerosis, glaucoma and brain tumours (Hanuš, 2009). The hemp seed also contains α-, β- and γ- tocopherols, where the proportion of gamma tocopherols are higher than that of other two, and both gamma and alpha tocopherols have shown to have anti-inflammatory properties in injuries where inflammation takes place (Crescente et al., 2018). The pharmacological and physiological study of hemp seed has shown that its actions are organ-specific which regulate the taken food, gastric secretion, gastrointestinal mobility, visceral sensation and intestinal inflammation of the gut (Bapat, 2015).

Immunomodulatory
The immunomodulatory function of tetrahydrocannabinol has been examined in the laboratory on the animal having multiple sclerosis and encephalomyelitis, where the death is observed in animal treated with placebo but not in the animals treated with tetrahydrocannabinol. Moreover, slight or no inflammatory reaction has been observed (ElSohly, 2007). Hemp protein contains all nine essential amino acids which are required by the human body where arginine and glutamic acid are present in larger quantity while sulphur-containing amino acids are present in moderate amount. Clinical studies of arginine show that it is beneficial in detoxification of ammonia, fetal development and decreasing the resistance of insulin. Also, glutamate is essential owing to its crucial role as a neurotransmitter in the brain (Leonard et al., 2020).

Lipopolysaccharides (LPS) of hemp seed stimulate the glial cell of the brain and spinal cord which induces for the activation of nuclear factor kappa B, where the activated kappa B upregulate the pro-inflammatory enzymes and other mediators of inflammation (Leonard et al., 2020). In animal, the hemp seeds active ingredient THC and similar compounds administration have been found to have made a significant difference to many hormonal systems such as gonadal steroids, prolactin, growth and thyroid hormones and to activate the hypothalamic-pituitary-adrenal axis. However, in human similar results are not consistent due to increase in tolerance (EFSA, 2011; Hanuš, 2009). Hemp seed (endocannabinoid system), is considered to be the pertinent modulator of physiological activities of not only the central nervous system but also in the autonomic nervous system such as in the gastrointestinal tract, reproductive system, cardiovascular system, immune system and endocrine network (ElSohly, 2007; Gerra et al., 2010).

Neuroprotective
CBD is one of the psychoactive compounds of hemp seed which has been used for the treatment of Alzheimer’s disease and Parkinson’s disease (Fathordoobady et al., 2019; Hanuš, 2009; Leonard et al., 2020). It is also reported that when the diet is rich with hemp seed then it guards against neurodegenerative disease (Leonard et al., 2020). Tetrahydrocannabinol and other ingredients of hemp, in addition to other benefits, it is also found to be neuroprotective (Bapat, 2015; Baron, 2018). Some researchers have reported that when the rats were on diet with hemp seed, then they have shown noticeable longevity, the performance of memory was better and anti-ageing genes were also expressed. So many studies in case of human, have shown positive results of hemp seed diet in the treatment of neurologic disorders (EFSA, 2011; Leonard et al., 2020). Hemp was used not only for the therapeutics of earache but also for reducing neurological discomfort (Lozano, 2001). The potency of hemp is highly appreciable for the therapeutics of chronic neuropathic pain which is partially assigned by the endocannabinoid system modification of the descending supraspinal inhibitory pathways where these pathways get frequently damage in chronic pain syndromes (Baron, 2018; Crescente et al., 2018). In addition to endocannabinoid, a large number of other groups have been used as neuroprotective agents (Hanuš, 2009). Recently, Canada agreed a drug officially named Sativex A, consisting of cannabis extracts can be used as supplementary for the therapeutics of neuropathic pain related to multiple sclerosis (ElSohly, 2007; Gerra et al., 2010). In addition to this, a large number of cannabis by-products have been used for the therapeutics of Central Nervous System disorder, neurodegenerative disorder and psychiatric disorders and have been patented and published (ElSohly, 2007; Gerra et al., 2010; Hanuš, 2009; Leizer et al., 2000; Wang et al., 2008).

Cardio-protective
Hemp seed is rich in essential fatty acids and PUFAs, in addition to protein, vitamin, carbohydrate, and minerals and when essential fatty acids are deficient in the cell then atherogenic process occurs. Hence, hemp seed is considered to be one of the suitable candidates for anti-atherosclerotic result and also helps in lowering cholesterol and high blood pressure. Higher utilization of PUFAs is directly related to a decrease in risks for cardiovascular disease (Latif & Anwar, 2009; Leizer et al., 2000; Leonard et al., 2020; Seo et al., 2012). Phytosterols present in hemp seed also reduces low-density lipoprotein in serum which directly helps in preventing cardiovascular disease as well as atherosclerosis (Seo et al., 2012). Hemp seed oil is also considered to be essential supplements for the treatment of coronary heart disease (Leonard et al., 2020). The ratio of omega-6 to omega-3 in hemp seed oil is 2:1 to 3:1 which is crucial properties of hemp seed oil and helps in maintaining various physiological mechanisms including vasodilation (Fathordoobady et al., 2019).

Recently, the endocannabinoid system has gained recognition as a significant regulator of various physiological functions within the body’s different systems, including the cardiovascular system (ElSohly, 2007; Gerra et al., 2010; Brierley et al., 2023). In hemp seed, the amino acids arginine is found to be in high levels which work as a precursor for the formation of nitric oxide which helps in the signalling pathway of the cardiovascular system and maintains homeostasis of blood pressure (Crescente et al., 2018). It is also reported that supplementary hemp seed can be used as anti-thrombotic and anti-vasoconstrictive (Leizer et al., 2000). Additionally, hemp seed free from THC has been used as one of the supplements for lowering the level of plasma cholesterol in rats (Saberivand et al., 2010).

Premenstrual syndrome (PMS) and menopause
PMS is an uneasiness in women during their reproductive period before the menstruation. This decreases with increase in age and dismissed at menopause (Freeman et al., 2004). The use of hemp seed as a medicine in obstetrics and gynaecology from the ancient time has been restudied. It was used as a supplement for the herbal formulation of headaches, insomnia, agoraphobia, depression, uterine haemorrhage and gastroenteritis. The phytoestrogens such as genistein from hemp cause an increase in the level of high-density lipoprotein cholesterol (HDL-C) and decrease in the level of low-density lipoprotein cholesterol (LDL-C) (Liu et al., 2023). The women who are in the proximity of menopause have decreased estrogen level because of which it causes depression, sleep disturbances, anxiety, irritability, cognitive dysfunction and panic disorders. Hence, the estrogens from hemp seed help to enhance the well-being, mood and better feelings as well prevent all the disturbances occurs because of deficiency of estrogens (Saberivand et al., 2010).

THC from hemp seed has control over hypothalamus regions which in turns controls the neuroendocrine physiology. When there is a decreased level of norepinephrine release, it stimulates the CB1 receptors which result in a decreased level of gonadotropin-releasing hormone and suppress the release of follicle-stimulating hormone and luteinizing hormone from the pituitary gland. And finally have control on Premenstrual Syndrome (PMS) to a certain extent (ElSohly, 2007; Freeman et al., 2004; Jin et al., 2018; Mukhtar, Elbagir & Gubara, 2012).

Anticancer
Hemp seed contains a large number of phenolic compounds and they have shown the anti-cancer effects in a clinical trial, also in Canada certain products have been used for therapeutics of cancer pain (Baron, 2018; Crescente et al., 2018; Hanuš, 2009; Jin et al., 2018; Wang et al., 2008). As PUFAs are present in hemp seed and excessive intake of PUFAs are related to the reduction in the risk of cancer diseases (Leonard et al., 2020). The omega-3 fatty acid of hemp seed is necessary for both growth and development. In addition to this, it is used as therapeutic agent for cancer and other diseases like cardiovascular, autoimmune, etc. (Jeong et al., 2014; Leizer et al., 2000). Beta-sitosterol is one of the ingredients of hemp seed whose clinical trial shows that it can be used as a therapeutics to a certain extent for prostatic hyperplasia (Leonard et al., 2020). Gamma tocopherol as one of the ingredients of hemp seed largely has been used as therapeutics for cancer especially for colon cancer (Leizer et al., 2000).”