Testing for Betel Nut Alkaloids in Hair of Papuans Abusers using UPLC–MS/MS and UPLC–Q-Tof-MS

Abstract

Betel nut is the fruit of Areca palm, growing in Papua New Guinea. Mixed with limestone and stick mustard, arecoline and guvacoline, which are present in betel nut, are hydrolyzed into arecaidine and guvacine, respectively. As part of the study on dietary habits of Papuans residents, our laboratory was asked to analyze the four alkaloids in hair to document long-term exposure. Hair samples were collected from 19 adult subjects (males = 11; females = 8), by some of the authors, and were sent to the laboratory for analysis. The four alkaloids have very similar chemical structures. In order to accurately identify the drugs, two methods were developed. First, the compounds were identified using an ultra-high-performance liquid chromatography system coupled to time-of-flight mass spectrometry. Then, they were quantified by an ultra-high-performance liquid chromatography system coupled to tandem mass spectrometry. After decontamination with dichloromethane, hair samples were cut into very small segments and 20 mg were incubated in methanol for 2 h 30 min in an ultrasound bath. After cooling, the methanol was evaporated to dryness in presence of 20-μL octanol to prevent volatilization. Nicotine-d4 was used as an internal standard. Linearity was observed for concentrations ranging from the limit of quantification to 20 ng/mg for arecoline, arecaidine, guvacine and guvacoline. Measured concentrations were in the range 60 pg/mg to 18 ng/mg for arecoline (n = 19), 14 pg/mg to 2.5 ng/mg for guvacoline (n = 11), 63 pg/mg to 3.8 ng/mg for arecaidine (n = 11) and 100 pg/mg to 3.2 ng/mg for guvacine (n = 6). There was no correlation between concentrations of arecoline and arecaidine (ratio from 0.01 to 0.18) and guvacoline and guvacine (ratio from 0.06 to 3.50). However, the identification of these substances in hair is a good marker of consumption of betel nut and allows us to document a local practice that remains difficult to evaluate just by questioning.

Pharmacology of Herbal Sexual Enhancers: A Review of Psychiatric and Neurological Adverse Effects

“…The content of major alkaloids arecoline, arecaidine, guvacoline, and guvacine in the fresh nut is approximately 0.30–0.63%, 0.31–0.66%, 0.03–0.06%, and 0.19–0.72%, respectively, although it may vary with maturation [ 160 , 161 , 162 , 163 ]. Due to the presence of calcium oxide, arecoline and guvacoline are hydrolyzed to arecaidine and guvacine, respectively, in basic conditions [ 163 , 164 ]. Areca alkaloids mediate the autonomic responses of the parasympathetic nervous system and the synaptic transmission in the peripheral nervous system [ 164 , 165 , 166 ], and impact various aspects of brain function and regulation [ 167 , 168 ].…”

“…Due to the presence of calcium oxide, arecoline and guvacoline are hydrolyzed to arecaidine and guvacine, respectively, in basic conditions [ 163 , 164 ]. Areca alkaloids mediate the autonomic responses of the parasympathetic nervous system and the synaptic transmission in the peripheral nervous system [ 164 , 165 , 166 ], and impact various aspects of brain function and regulation [ 167 , 168 ]. Arecoline is a partial muscarinic (M) agonist and possesses a higher affinity for M receptors than guvacoline [ 24 , 169 ].…”

“…Ashwagandha has numerous applications in traditional Indian medicines and has been used for more than 5000 years as an aphrodisiac, antioxidant, antimicrobial, adaptogenic, diuretic, tonic, narcotic, immuno-stimulant, anti-inflammatory agent, anti-stress, antiulcer, among many other purposes. Positive effect in insomnia, anxiety, chronic stress, weight management, thyroid gland function, telomerase, cardio-respiratory endurance, muscle strength, recovery of male and female sexual function, and senescence was demonstrated in preclinical and clinical studies [ 164 ]. Excellent tolerability is generally reported after using ashwagandha for weeks at therapeutic doses, with statistically insignificant adverse effects compared to placebo [ 281 , 282 ].…”

Genome skims analysis of betel palms (Areca spp., Arecaceae) and development of a profiling method to assess their plastome diversity

“…Nowadays, as many as 600 million people worldwide, concentrated in the Indo-Pacific region, regularly chew betel nut. Seeds contain four major alkaloids (Gheddar et al, 2020) which are central nervous system stimulants and are highly addictive (acting on the same receptor proteins in the brain as nicotine). They have psychoactive effects and are appreciated for their appetite suppressant and antimycobacterial properties (Gautam et al, 2007).…”

The degradation of forchlorfenuron in the model kiwifruit juice by ultrasonic treatment

“…Due to the powerful resolution, high sensitivity and good separation, high‐performance liquid chromatography (HPLC), and ultra‐performance liquid chromatography‐Q‐time of flight mass spectrometry (UPLC‐Q‐TOF‐MS) have become favorable approaches in metabolism identification of drug, Chinese herbs and pesticides (Jiao et al, ; Wen & Zhu, ; Xiao, Meichen, Li, Yun, & Cheng, ; Xie, Zhong, & Chen, ). Rapid identification and analysis for metabolism profiles and chemical components of Xuanmai Ganjie Granules, Nanxing acesodyne plaster and betel nut alkaloids in hair, UPLC‐Q‐TOF‐MS was applied (Gheddar et al, ; Guo et al, ; Wu, Wang, Yang, & Sui, ). Hu and Li () disposed the CPPU residues in watermelon by solid‐phase extraction (SPE) and HPLC.…”