Cotinine ELISA KIT

Name: Cotinine ELISA KIT
SKU: DEIA04892
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

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COA

SDS

Datasheet

Sample

Serum, urine or saliva.

Species Reactivity

Human

Detection Method

cELISA

Intended Use

The Cotinine ELISA Kit is intended for the measurement of Cotinine in serum, urine or saliva.

Contents of Kit

1. Microwell coated with polyclonal Ab to Cotinine. 12 × 8
2. Standard Set (ready to use). 0.5 mL
3. Cotinine HRP Enzyme Conjugate (ready to use). 12 mL
4. TMB Substrate (ready to use). 12 mL
5. Stop Solution (ready to use). 12 mL
6. Wash Concentrate. 20×, 1 bottle, 25 mL

Storage

1. Store the kit at 2 - 8°C.
2. Keep microwells sealed in a dry bag with desiccants.
3. The reagents are stable until expiration of the kit.
4. Do not expose test reagents to heat, sun, or strong light.

Performance Characteristics

Accuracy
20 urine samples from non-smokers were screened with this Cotinine ELISA method. All 20 samples screened negative with the ELISA method. 15 samples from smokers which contained various amounts of Cotinine were screened with This Cotinine Direct ELISA Kit. All 15 samples showed a presence of Cotinine at a level greater than 500 ng/mL. Three urine samples submitted by individuals exposed to passive inhalation for over 30 days all showed levels of 5 to 10 ng/mL of Cotinine when extrapolated of a dose response curve.
Sensitivity
Assay sensitivity based on the minimum Cotinine concentration required to produce a three-standard deviation from assay Ao is 1 ng/mL.
Specificity
The specificity of this Cotinine ELISA was determined by generating inhibition curves for each of the compounds listed below the antisera cross-reactivity below.

Detection Range

5-100 ng/mL

Sensitivity

1 ng/mL

General Description

Exposure to tobacco smoke can be detected by measuring nicotine and its metabolites. Nicotine has a short half life and is not used as a marker for tobacco smoke exposure. Cotinine due to its longer half life has been used in research as a reliable marker for smoking status and smoking cessation studies. The Cotinine Direct ELISA Kit is designed for the detection Cotinine in serum and urine. It can also be adapted for other fluids.

Citations

Publication (3)

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Ameliorative Effect of Gum Acacia on Hookah Smoke-Induced Testicular Impairment in Mice

Ali BH, Al-Salam S, Balushi KAA, Za'abi MA, Adham SA, Beegam S, Yuvaraju P, Manoj P, Nemmar A

Biomolecules2020 MayPubMed ID:32414135Read Article

Applications: ELISA

Reactive species: Human

"Abstract: We investigated some reproductive actions of hookah smoke (HS) exposure (30 min/day, for 30 days) in male mice, and the possible mitigative effect of the prebiotic agent gum acacia (GA) thereon. Control mice were air-exposed (AE). Twenty-four hours after the last exposure, the levels of some plasma reproductive hormones, biochemical markers of inflammation, oxidative and nitrosative stress and testicular histopathology were assessed. The urinary level of cotinine, a major nicotine metabolite, was also measured. HS exposure induced significant decreases in testosterone, estradiol, luteinizing hormone, and androgen binding protein, as well as glutathione reductase activity and levels of nitrite and total nitrite. Plasma inhibin B, alkaline phosphatase, lipopolysaccharide binding protein, uric acid, lactate dehydrogenase, lipid peroxidation, 8-oxo-2'-deoxyguanosine, and cytochrome C were significantly increased following HS exposure. In testicular homogenate, nuclear factor-κB (NF-ĸB), nuclear factor erythroid 2-related factor 2 (Nrf2), interleukin- 6 (IL-6), interleukin-1β (IL-1β), transforming growth factor-β1(TGF- β1), and tumor necrosis factor-α (TNF- α) were all significantly elevated, and the steroidogenic acute regulatory protein (StAR) significantly decreased. Histopathologically, there was slight impairment and disorganization of spermatogenesis. Urinary cotinine concentration was elevated significantly in the HS-exposed group compared with the air-exposed group. GA co-administration mitigated the adverse actions of HS measured. In conclusion, daily exposure to HS at the above dose induced adverse actions on the reproductive system of male mice. GA co-administration significantly mitigated these effects by reducing the inflammation, oxidative and nitrosative stress, via a mechanism involving Nrf2, and reduction of StAR expression."

"Article snippet: The urinary level of cotinine was assayed by an ELISA kit from Creative Diagnostics (Shirley, NY, USA)."

Urine cotinine level of mice exposed to air (AE) or hookah smoke (HS) with or without GA treatment. (Ali BH, <i>et al</i>, 2022)

Figure 1. Urine cotinine level of mice exposed to air (AE) or hookah smoke (HS) with or without GA treatment.

Chronic Water-Pipe Smoke Exposure Induces Injurious Effects to Reproductive System in Male Mice

Badreldin H Ali, Khalid A Al Balushi, Mohammed Ashique, Asem Shalaby, Mohammed A Al Kindi, Sirin A Adham, Turan Karaca, Sumaya Beegam, Priya Yuvaraju, Abderrahim Nemmar

Front Physiol2017 AprPubMed ID: 28420996Read Article

Applications: ELISA

Reactive species: Human

"Abstract: There is a global increase in the popularity of water-pipe tobacco smoking including in Europe and North America. Nevertheless, little is known about the male reproductive effects of water-pipe smoke (WPS), especially after long-term exposure. Here, we assessed effects of WPS exposure (30 min/day) in male mice for 6 months. Control mice were exposed to air-only for the same period of time. Twenty-four hours after the last exposure, testicular histopathology, and markers of inflammation and oxidative stress, and the tyrosine–protein kinase vascular endothelial growth factor receptor 1 (VEGFR1) were assessed in testicular homogenates. Moreover, plasma testosterone, estradiol, and luteinizing hormone (LH) concentrations were also measured. Chronic WPS exposure induced a significant decrease of testosterone and estradiol, and a slight but significant increase of LH. Glutathione reductase, catalase, and ascorbic acid were significantly decreased following WPS exposure. Plasma concentration of leptin was significantly decreased by WPS exposure, whereas that of tumor necrosis factor α and interleukin 6 was significantly increased. Histopathological analysis of the testes revealed the presence of a marked reduction in the diameter of the seminiferous tubules with reduced spermatogenesis. Transmission electron microscopy examination showed irregular thickening and wrinkling of the basement membranes with abnormal shapes and structures of the spermatozoa. VEGFR1 was overexpressed in the testis of the mice exposed to WPS and was not detected in the control. The urine concentration of cotinine, the predominant metabolite of nicotine, was significantly increased in the WPS-exposed group compared with the control group. "

"Article snippet: The concentrations of cotinine in urine (ELISA kit from Creative Diagnostics, Shirley, NY, USA) were measured after the last exposure to either WPS or air on urine samples collected overnight using metabolic cages for urine collection."

The concentrations of cotinine in plasma (A) and urine (B) following air exposure (AE) of control mice (open columns), and mice exposed to water-pipe smoke (WPS) for 30 min per day for 6 months (filled columns). (Badreldin H Ali, <i>et al</i>, 2017)

Figure 1. The concentrations of cotinine in plasma (A) and urine (B) following air exposure (AE) of control mice (open columns), and mice exposed to water-pipe smoke (WPS) for 30 min per day for 6 months (filled columns).

Waterpipe Tobacco Smoke Inhalation Triggers Thrombogenicity, Cardiac Inflammation and Oxidative Stress in Mice: Effects of Flavouring

Abderrahim Nemmar, Suhail Al-Salam, Sumaya Beegam, Priya Yuvaraju, Nur Elena Zaaba, Javed Yasin, Badreldin H Ali

Int J Mol Sci2020 FebPubMed ID: 32075078Read Article

Applications: ELISA

Reactive species: Human

"Abstract: The consumption of water-pipe smoking (WPS) has been promoted by the use of flavoured tobacco. However, little is known about the impact of flavouring on the cardiovascular toxicity induced by WPS inhalation. Here, we compared the cardiovascular effects and underlying mechanism of actions of plain (P) (unflavoured) versus apple-flavoured (AF) WPS (30 minutes/day, 5 days/week for 1 month) in mice. Control mice were exposed to air. Both P- and AF-WPS inhalation induced an increase in systolic blood pressure, thrombogenicity and plasma concentration of fibrinogen and von Willebrand factor. In heart homogenates, AF-WPS inhalation caused an increase of 8-isoprostane and a decrease in the levels of reduced glutathione (GSH) and superoxide dismutase (SOD). Nevertheless, P-WPS decreased only the activity of SOD. The concentrations of tumour necrosis factor α and interleukin 1β were increased only in heart homogenates of mice exposed to AF-WPS. "

"Article snippet: The quantiﬁcation of urinary cotinine concentrations was performed using an ELISA kit purchased from Creative Diagnostics (Shirley, NY, USA)."

Eﬀects of plain water-pipe smoke (P-WPS) or apple-ﬂavoured (AF)-WPS on cotinine concentrations in urine. (Abderrahim Nemmar, <i>et al</i>, 2020)

Figure 1. Eﬀects of plain water-pipe smoke (P-WPS) or apple-ﬂavoured (AF)-WPS on cotinine concentrations in urine.

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Elevated plasma cotinine is associated with an increased risk of developing IBD, especially among users of combusted tobacco

PLOS ONE

Authors: Widbom, Lovisa; Schneede, Jorn; Midttun, Oivind; Ueland, Per Magne; Karling, Pontus; Hultdin, Johan

Abstract

Objective Smoking has previously been associated with inflammatory bowel disease (IBD), but no study has reported on cotinine, an objective, biochemical measure of tobacco use. We aimed at testing the hypothesis that cotinine levels among healthy subjects are associated with an increased risk of developing IBD in later life. Design We analysed plasma cotinine and evaluated corresponding lifestyle questionnaires that included tobacco habits in subjects (n = 96) who later developed late-onset IBD (70 ulcerative colitis (UC) and 26 Crohn's disease (CD)) and in sex and age-matched controls (n = 191). Results Patients who later developed IBD had significantly higher plasma cotinine levels compared to controls. In multivariable analysis, higher log-cotinine was associated with a higher risk of developing IBD (OR 1.34 (95% CI 1.01-1.63)). After stratifying for time to diagnosis, the association was only significant in subjects with shorter time (< 5.1 years) to diagnosis (OR 1.45 (1.09-1.92)). The findings were similar for UC- and CD-cases, but did not reach statistical significance in CD-cases. Although plasma cotinine concentrations were higher in snuff users compared to combusted tobacco users, no increase in the risk of IBD and lower risk of developing IBD among subjects with shorter time (< 5.1 years) to diagnosis was seen among snuff users. Conclusions Cotinine, a biomarker of tobacco use, is associated with increased risk of developing late-onset IBD in general, and UC in particular. No increased risk among snuff users indicates that other components in combusted tobacco than nicotine may be involved in the pathogenesis of IBD among smokers.

Exposure to perfluoroalkyl substances (PFAS) and associations with thyroid parameters in First Nation children and youth from Quebec

ENVIRONMENT INTERNATIONAL

Authors: Caron-Beaudoin, Elyse; Ayotte, Pierre; Sidi, Elhadji Anassour Laouan; McHugh, Nancy Gros-Louis; Lemire, Melanie

Abstract

Background: Perfluoroalkyl substances (PFASs) are found in several consumer goods. Exposure to PFASs in children has been associated with alteration in thyroid hormones, which have critical roles in brain function. Objective: In 2015, 198 children and youth (3-19y) were recruited as part of the pilot project Jeunes, Environnement et Sante/Youth, Environment and Health (JES!-YEH!), realized in collaboration with four First Nation communities in Quebec. We aimed to evaluate serum concentrations of PFASs in relation to concentrations of thyroid-stimulating hormone (TSH), free thyroxine (T4) and thyroglobulin while adjusting for relevant confounders. Methods: PFASs (PFOS, PFOA, PFHxS, PFNA), 2,2',4,4'-Tetrabromodiphenyl ether (PBDE-47) thyroid parameters (TSH, free T4, and thyroglobulin) were measured in serum samples of 186 participants. Iodine, creatinine, and cotinine were measured in urine samples. Serum levels of PFASs were compared to those measured in the general Canadian population and elsewhere. Multivariate regression analyses were performed to determine associations between PFASs and TSH, free T4 and thyroglobulin. Results: PFOS, PFOA and PFHxS serum concentrations were low. However, PFNA concentrations among participants aged 12 to 19 years old from Anishinabe communities were three times higher than those measured in the Canadian Health Measures Survey (2009-2011) for the same age group (Geometric Means: 3.01 mu g/L and 0.71 mu g/L, respectively) and were particularly higher in the Anishinabe participants aged 6 to 11 years old (GM: 9.44 mu g/L). Few participants had levels of TSH, free T4, and thyroglobulin outside age-specific paediatric ranges. When adjusted for relevant covariates and other contaminants, PFNA serum concentrations were positively associated with free T4 levels (Adjusted beta = 0.36; p = 0.0014), but not with TSH and thyroglobulin levels. No association was observed between the other PFAS and thyroid hormones parameters. Conclusion: This pilot project reveals among the highest exposure to PFNA in children reported until today, and suggests effects of PFNA as an endocrine disruptor, highlighting the importance of investigating the sources and effects of disproportionate exposure to emerging contaminants in some indigenous communities and ban all PFAS at the international scale.