Nitrate INFORMER Nitrosamine Study

NCT ID: NCT05045807

Last Updated: 2022-04-01

Study Results

Results pending

The study team has not published outcome measurements, participant flow, or safety data for this trial yet. Check back later for updates.

Basic Information

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Recruitment Status

COMPLETED

Clinical Phase

NA

Total Enrollment

6 participants

Study Classification

INTERVENTIONAL

Study Start Date

2021-10-11

Study Completion Date

2021-12-31

Brief Summary

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Nitrate is a controversial component of vegetables, meat, and drinking water. The now well-established benefits of nitrate, through the enterosalivary nitrate-nitrite-nitric oxide (NO) pathway, on cardiovascular risk factors and long-term cardiovascular disease risk are tarnished by a continuing concern about a link between nitrate ingestion and cancer. This can result in misguided advice to avoid consumption of high-nitrate leafy green vegetables by both the media and the scientific literature. A recent media headline stated, "Cancer alert over rocket: trendy salad leaves exceed safe levels of carcinogenic nitrates in one in every ten samples". One scientific review stated, "the presence of nitrate in vegetables, as in water and generally in other foods, is a serious threat to man's health". Controversy in the literature, and gaps in the knowledge are leading to confusing messages around vegetables that may play a critical role in cardiovascular health.

The major dietary sources of nitrate are vegetables, meat, and drinking water. Source of nitrate could be a crucial factor determining whether the consumption of nitrate is linked with beneficial (such as improving cardiovascular health) versus harmful (N-nitrosamine formation) effects. For example, unlike meat and water-derived nitrate, vegetables contain high levels of vitamin C and/or polyphenols that may inhibit the production of N-nitrosamines. So far, no study has investigated the formation of N-nitrosamines after consumption of these different sources in humans.

A pilot study will be conducted to determine that endogenous N-nitrosamine formation is observed after ingestion of meat with added nitrate and nitrite and that consumption of vegetables with the meat containing added nitrate and nitrite will inhibit the production of N-nitrosamines.

Detailed Description

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Study design:

A crossover study design will be used with a 1-week washout period. Each participant will complete the scheduled study visits for one of the dietary interventions which will be assigned in random order. Following the washout, the participants will then complete the scheduled study visits for the remaining intervention. Participants will be provided with a low nitrate and N-nitrosamine meal on the preceding evening of each study visit. These meals will be consistent across all study visits. Participants will be asked to refrain from drinking coffee and any alcoholic beverage and doing any exercise 24 hours prior to the study visit.

Dietary interventions:

Meat with added nitrate: Prosciutto/pancetta/Parma ham/salami (all derived from pork) prepared by a commercial butcher with sodium nitrate as an additive.

Meat with added nitrate plus vegetables: The same intervention described above consumed together with mixed vegetables.

Assessments:

At each clinic visit, baseline samples of urine will be collected for measurement of N-nitrosamines, nitrate, and nitrite. After the intervention, all urine within the first 240 minutes and then subsequent 20-hour period will be collected for measurement of N-nitrosamines, nitrate, and nitrite.

Conditions

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Health Risk Behaviors

Study Design

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Allocation Method

RANDOMIZED

Intervention Model

CROSSOVER

prospective, mono-centre, randomised, controlled, crossover study
Primary Study Purpose

PREVENTION

Blinding Strategy

SINGLE

Outcome Assessors
Given the nature of the interventions, participants, and the investigators responsible for delivering the interventions will be unblinded throughout the trial. However, all researchers performing the laboratory analyses and data analyses will be blinded to the interventions that the participants received until after the data analysis has been performed.

Study Groups

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Meat with added nitrate

Prosciutto/pancetta/Parma ham/salami (all derived from pork) prepared by a commercial butcher with sodium nitrate as an additive.

Group Type EXPERIMENTAL

Meat with added nitrate

Intervention Type DIETARY_SUPPLEMENT

Prosciutto/pancetta/Parma ham/salami (all derived from pork) prepared by a commercial butcher with the sodium nitrate and nitrite as an additive.

Meat with added nitrate plus vegetables

The same intervention as of Arm "meat with added nitrate" consumed together with mixed vegetables.

Group Type EXPERIMENTAL

Meat with added nitrate plus vegetables

Intervention Type DIETARY_SUPPLEMENT

The same intervention as of intervention "meat with added nitrate" consumed together with mixed vegetables.

Interventions

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Meat with added nitrate

Prosciutto/pancetta/Parma ham/salami (all derived from pork) prepared by a commercial butcher with the sodium nitrate and nitrite as an additive.

Intervention Type DIETARY_SUPPLEMENT

Meat with added nitrate plus vegetables

The same intervention as of intervention "meat with added nitrate" consumed together with mixed vegetables.

Intervention Type DIETARY_SUPPLEMENT

Eligibility Criteria

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Inclusion Criteria

* Six healthy, ambulant, community-dwelling men and women aged between 18 to 55 years old and with no history of major chronic disease will be recruited from the Perth general population.

Exclusion Criteria

Individuals volunteering to participate in the study will be excluded according to the following criteria:

* current or recent (\<12 months) smoking
* body mass index (BMI) \<18 or \> 35 kg/m2
* systolic blood pressure \> 160 mmHg
* diastolic blood pressure \> 100 mmHg
* any major illness such as cancer, psychiatric illness, diagnosed diabetes
* use of any of the following medications: statins, antihypertensives, nitric oxide donors, antithrombotic medication, anti-coagulant medication, anti-arrhythmic drugs, beta-blockers, regular aspirin use, regular proton pump inhibitor use
* alcohol consumption \> 30g/day
* who are pregnant, lactating, or wishing to become pregnant during the study
* use of antibiotics within the previous 12 weeks of the study
* regular use of mouthwash and not willing to cease mouthwash use for the duration of the study
* participation on other research studies
* major gastrointestinal tract condition e.g. Crohns disease and inflammatory bowel disease
* and inability or unwillingness to follow the study protocol.
Minimum Eligible Age

18 Years

Maximum Eligible Age

55 Years

Eligible Sex

ALL

Accepts Healthy Volunteers

Yes

Sponsors

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The University of Western Australia

OTHER

Sponsor Role collaborator

Flinders University

OTHER

Sponsor Role collaborator

Edith Cowan University

OTHER

Sponsor Role lead

Responsible Party

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Responsibility Role SPONSOR

Principal Investigators

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Catherine P Bondonno, PhD, RNutr.

Role: PRINCIPAL_INVESTIGATOR

Edith Cowan University

Locations

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Royal Perth Hospital Research Foundation

Perth, Western Australia, Australia

Site Status

Countries

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Australia

References

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Lundberg JO, Weitzberg E. NO-synthase independent NO generation in mammals. Biochem Biophys Res Commun. 2010 May 21;396(1):39-45. doi: 10.1016/j.bbrc.2010.02.136.

Reference Type RESULT
PMID: 20494108 (View on PubMed)

Blekkenhorst LC, Bondonno NP, Liu AH, Ward NC, Prince RL, Lewis JR, Devine A, Croft KD, Hodgson JM, Bondonno CP. Nitrate, the oral microbiome, and cardiovascular health: a systematic literature review of human and animal studies. Am J Clin Nutr. 2018 Apr 1;107(4):504-522. doi: 10.1093/ajcn/nqx046.

Reference Type RESULT
PMID: 29635489 (View on PubMed)

Bondonno CP, Blekkenhorst LC, Liu AH, Bondonno NP, Ward NC, Croft KD, Hodgson JM. Vegetable-derived bioactive nitrate and cardiovascular health. Mol Aspects Med. 2018 Jun;61:83-91. doi: 10.1016/j.mam.2017.08.001. Epub 2017 Sep 7.

Reference Type RESULT
PMID: 28802834 (View on PubMed)

Spiegelhalder B, Eisenbrand G, Preussmann R. Influence of dietary nitrate on nitrite content of human saliva: possible relevance to in vivo formation of N-nitroso compounds. Food Cosmet Toxicol. 1976 Dec;14(6):545-8. doi: 10.1016/s0015-6264(76)80005-3. No abstract available.

Reference Type RESULT
PMID: 1017769 (View on PubMed)

Tannenbaum SR, Weisman M, Fett D. The effect of nitrate intake on nitrite formation in human saliva. Food Cosmet Toxicol. 1976 Dec;14(6):549-52. doi: 10.1016/s0015-6264(76)80006-5. No abstract available.

Reference Type RESULT
PMID: 1017770 (View on PubMed)

Gangolli SD, van den Brandt PA, Feron VJ, Janzowsky C, Koeman JH, Speijers GJ, Spiegelhalder B, Walker R, Wisnok JS. Nitrate, nitrite and N-nitroso compounds. Eur J Pharmacol. 1994 Nov 1;292(1):1-38. doi: 10.1016/0926-6917(94)90022-1.

Reference Type RESULT
PMID: 7867685 (View on PubMed)

Mirvish SS. Role of N-nitroso compounds (NOC) and N-nitrosation in etiology of gastric, esophageal, nasopharyngeal and bladder cancer and contribution to cancer of known exposures to NOC. Cancer Lett. 1995 Jun 29;93(1):17-48. doi: 10.1016/0304-3835(95)03786-V.

Reference Type RESULT
PMID: 7600541 (View on PubMed)

IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. IARC monographs on the evaluation of carcinogenic risks to humans. Ingested nitrate and nitrite, and cyanobacterial peptide toxins. IARC Monogr Eval Carcinog Risks Hum. 2010;94:v-vii, 1-412. No abstract available.

Reference Type RESULT
PMID: 21141240 (View on PubMed)

Hord NG, Tang Y, Bryan NS. Food sources of nitrates and nitrites: the physiologic context for potential health benefits. Am J Clin Nutr. 2009 Jul;90(1):1-10. doi: 10.3945/ajcn.2008.27131. Epub 2009 May 13.

Reference Type RESULT
PMID: 19439460 (View on PubMed)

Blekkenhorst LC, Prince RL, Ward NC, Croft KD, Lewis JR, Devine A, Shinde S, Woodman RJ, Hodgson JM, Bondonno CP. Development of a reference database for assessing dietary nitrate in vegetables. Mol Nutr Food Res. 2017 Aug;61(8). doi: 10.1002/mnfr.201600982. Epub 2017 May 3.

Reference Type RESULT
PMID: 28105786 (View on PubMed)

Bartsch H, Ohshima H, Pignatelli B. Inhibitors of endogenous nitrosation. Mechanisms and implications in human cancer prevention. Mutat Res. 1988 Dec;202(2):307-24. doi: 10.1016/0027-5107(88)90194-7.

Reference Type RESULT
PMID: 3057363 (View on PubMed)

Levallois P, Ayotte P, Van Maanen JM, Desrosiers T, Gingras S, Dallinga JW, Vermeer IT, Zee J, Poirier G. Excretion of volatile nitrosamines in a rural population in relation to food and drinking water consumption. Food Chem Toxicol. 2000 Nov;38(11):1013-9. doi: 10.1016/s0278-6915(00)00089-2.

Reference Type RESULT
PMID: 11038239 (View on PubMed)

Bartholomew B, Hill MJ. The pharmacology of dietary nitrate and the origin of urinary nitrate. Food Chem Toxicol. 1984 Oct;22(10):789-95. doi: 10.1016/0278-6915(84)90116-9.

Reference Type RESULT
PMID: 6541617 (View on PubMed)

Bondonno CP, Croft KD, Puddey IB, Considine MJ, Yang X, Ward NC, Hodgson JM. Nitrate causes a dose-dependent augmentation of nitric oxide status in healthy women. Food Funct. 2012 May;3(5):522-7. doi: 10.1039/c2fo10206d. Epub 2012 Feb 16.

Reference Type RESULT
PMID: 22336776 (View on PubMed)

Bondonno CP, Downey LA, Croft KD, Scholey A, Stough C, Yang X, Considine MJ, Ward NC, Puddey IB, Swinny E, Mubarak A, Hodgson JM. The acute effect of flavonoid-rich apples and nitrate-rich spinach on cognitive performance and mood in healthy men and women. Food Funct. 2014 May;5(5):849-58. doi: 10.1039/c3fo60590f.

Reference Type RESULT
PMID: 24676365 (View on PubMed)

Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJ, Holmes SP. DADA2: High-resolution sample inference from Illumina amplicon data. Nat Methods. 2016 Jul;13(7):581-3. doi: 10.1038/nmeth.3869. Epub 2016 May 23.

Reference Type RESULT
PMID: 27214047 (View on PubMed)

Cole JR, Wang Q, Fish JA, Chai B, McGarrell DM, Sun Y, Brown CT, Porras-Alfaro A, Kuske CR, Tiedje JM. Ribosomal Database Project: data and tools for high throughput rRNA analysis. Nucleic Acids Res. 2014 Jan;42(Database issue):D633-42. doi: 10.1093/nar/gkt1244. Epub 2013 Nov 27.

Reference Type RESULT
PMID: 24288368 (View on PubMed)

Parks DH, Chuvochina M, Waite DW, Rinke C, Skarshewski A, Chaumeil PA, Hugenholtz P. A standardized bacterial taxonomy based on genome phylogeny substantially revises the tree of life. Nat Biotechnol. 2018 Nov;36(10):996-1004. doi: 10.1038/nbt.4229. Epub 2018 Aug 27.

Reference Type RESULT
PMID: 30148503 (View on PubMed)

Other Identifiers

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2021-02629-BONDONNO(pilot)

Identifier Type: -

Identifier Source: org_study_id

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