In vitro cytotoxicity of Nicotiana gossei leaves, used in the Australian Aboriginal smokeless tobacco known as pituri or mingkulpa

Moghbel, Nahid; Ryu, BoMi; Cabot, Peter J.; Steadman, Kathryn J.

doi:10.1016/j.toxlet.2016.05.011

Cited by 17 publications

(20 citation statements)

References 26 publications

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“…µg/g) and NNK (6.91 µg/g) for N. gossei leaves, which reflects the levels in the final product, pituri, are consistent with smokeless tobacco products in use around the world. The presence of other alkaloids and carcinogenic NNN and NNK is in line with the observed higher cytotoxicity of these products rather that attributed to nicotine alone (Moghbel et al, 2016) (Chapter 5).…”

Section: Discussionsupporting

confidence: 56%

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Investigations into the pituri plant: nicotine content, nicotine conversion to nornicotine, nicotine release and cytotoxicity of Australian native Nicotiana spp.

Moghbel¹

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The Aboriginal population of Central Australia use endemic Nicotiana spp. to make a smokeless tobacco product known as pituri that they chew/suck for nicotine absorption. This thesis describes the relative abundance of nicotine alkaloids amongst Australian Nicotiana spp., with special focus on the molecular characteristics of nicotine to nornicotine conversion.The most popular chewed species, N. gossei, is investigated for nicotine release and cytotoxicity in comparison to similar products to gain insight into potential hazards to pituri users.To analyse the alkaloids of Nicotiana leaves, a HPLC-UV method was developed to separate and quantify six closely related alkaloids (nicotine, nornicotine, anatabine, anabasine, myosmine, cotinine). A C18 column with a mobile phase of ammonium formate buffer (pH 10.5) separated the six alkaloids within 13 min with detection at 260 nm. Linearity, precision and reproducibility were satisfactory. The limit of quantification was 2.8 and 4.8 µg/mL for nornicotine and nicotine, respectively, and below 2 µg/mL for other alkaloids. This method quantifies more alkaloids and in less time than previously reported methods.Tobacco alkaloids are responsible in formation of carcinogenic tobacco specific nitrosamines such as N'-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). To quantify NNN and NNK, a fast LC-MS/MS method was developed using a HILIC column with a triple quadrupole tandem mass spectrometry. The linearity, accuracy, recovery and repeatability for this method were satisfactory, with quantification limit of 2.6 and 4.3 ng/mL for NNN and NNK, respectively. Nornicotine results from demethylation of nicotine, and is associated with negative effects on health. A group of cytochrome P450 genes that are mainly expressed during senescence or drying is involved in this conversion. The conversion loci were amplified in all studied 24 taxa, and sequenced in 6 selected species that contrast in conversion phenotype. Transcript accumulation of the responsible loci in fresh versus dried leaves of low or nonconverter N. gossei, N. excelsior and N. benthamiana maintained a steady level or a slight increase, but increased by 3 fold in cured leaves of the high converter N. goodspeedii, N. velutina and N. cavicola. This indicates the presence of functional loci that are triggered by curing only in high ii converter species and poses a potential risk for chewers of these species due to their greater potential for nornicotine production.The release of nicotine from the leaves of N. gossei was compared to that from a Swedish snus, the CORESTA reference smokeless tobacco (CRP2), and Nicabate chewing gum. A model buccal cavity system was developed and three different chewing conditions, performed manually were tested over 120 minutes: no chewing action, initial chewing and chewing at 15-minute intervals. To simulate the effect of alkaline wood ash, the effect of alkaline pH on the release of nicotine from N. gossei dry leaves was also evaluated. Samples ...

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Section: Discussionsupporting

confidence: 56%

“…The media in the vessels with maximum chewing action for all tested products was quantified for two major TSNAs, NNN and NNK following the validated LC-MS/MS method reported previously as mentioned in chapter 2 (Moghbel et al, 2016).…”

Section: Tsnas Analysis With Lc-ms/msmentioning

confidence: 99%

Investigations into the pituri plant: nicotine content, nicotine conversion to nornicotine, nicotine release and cytotoxicity of Australian native Nicotiana spp.

Moghbel¹

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“…However, one study suggested other biologically active compounds like NNN, NNK, etc., other than nicotine present in tobacco leave extract to be the source of cytotoxicity. 24 Another probable factor for the decrease in viability of the tuibur treatment groups would be the change in pH of the culture media. As we have shown in our result, the pH of both the two grades of tuibur are alkaline in nature, a slight rise in pH of the culture media was observed after the addition of both the tuibur (data not shown).…”

Section: Discussionmentioning

confidence: 99%

In vitro effect of tuibur (tobacco brew) on the viability of human blood lymphocytes

Lalruatfela

Zoremsiami

Jagetia

2017

Sci Vis

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The use of tobacco and its products are known to cause many illnesses including cancer. A smokeless tobacco locally manufactured called tuibur (tobacco brew) has been consumed by the Mizos from a very long time. In this experiment we aim to determine the cytotoxicity of tuibur by an in vitro study on tuibur-treated human peripheral blood lymphocytes. We have found that 24 h treatment of human lymphocytes with two grades of commercial tuibur and nicotine showed a concentration dependent decrease in cell viability. We, therefore, concluded that as the in vitro use of tuibur has an adverse effect on cell survival, its consumption might have potential side effects on the health of the users.

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“…It should be mentioned that recreational use is not recommended because the compounds are carcinogenic and can cause complications varying from vomiting and headache to severe depression. The chemistry of this species is complex and has both phytotoxicity and zootoxicity (Buta et al ., ; Severson et al ., ; Bentz & Neal, ; Moghbei et al ., ). However, pituri species should perhaps be studied for medicinally beneficial compounds.…”

Section: Platementioning

confidence: 99%

888. Nicotiana Gossei

Chase¹,

Christenhusz²

2018

Curtis's Botanical Magazine

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In vitro cytotoxicity of Nicotiana gossei leaves, used in the Australian Aboriginal smokeless tobacco known as pituri or mingkulpa

Cited by 17 publications

References 26 publications

Investigations into the pituri plant: nicotine content, nicotine conversion to nornicotine, nicotine release and cytotoxicity of Australian native Nicotiana spp.

Investigations into the pituri plant: nicotine content, nicotine conversion to nornicotine, nicotine release and cytotoxicity of Australian native Nicotiana spp.

In vitro effect of tuibur (tobacco brew) on the viability of human blood lymphocytes

888. Nicotiana Gossei

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