Nitrate INFORMER Nitrosamine Study

NCT05045807 | Completed

• 1 other identifier: 2021-02629-BONDONNO(pilot)
• Study Type: interventional
• Target: 6
• Locations: 1 country
• Sites: 1

Brief Summary

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.

Conditions

Health Risk Behaviors

Outcome Measures

Primary Outcomes (3)

• N-nitrosamines in urine pre each intervention

  Participants will be provided with a sterilized container and instructions to collect the first urine sample of the day which will be brought into the clinic. Urine aliquots will be frozen at -80°C until analysis. Level of N-nitrosamines will be measured by gas chromatography mass spectrometry (GCMS).

  At each clinic visit, baseline samples of urine will be collected.

• N-nitrosamines in urine post each intervention up to 240 min

  Participants will be provided with a sterilized container and instructions to collect all urine until 4 hours post intervention. Urine aliquots will be frozen at -80°C until analysis. Level of N-nitrosamines will be measured by gas chromatography mass spectrometry (GCMS).

  At each clinic visit, all urine samples within the first 240 min will be collected.

• N-nitrosamines in urine post each intervention after 4 hours up till 24 hours

  Participants will be provided with a sterilized container and instructions to collect all urine from 4 hours until 24 hours post intervention. Urine aliquots will be frozen at -80°C until analysis. Level of N-nitrosamines will be measured by gas chromatography mass spectrometry (GCMS).

  After each clinic visit, all urine from 4-hour to 24-hour period will be collected.

Study Arms (2)

Meat with added nitrate

EXPERIMENTAL

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

Dietary Supplement: Meat with added nitrate

Meat with added nitrate plus vegetables

EXPERIMENTAL

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

Dietary Supplement: Meat with added nitrate plus vegetables

Interventions

Meat with added nitrate 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

Meat with added nitrate plus vegetables DIETARY_SUPPLEMENT

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

Meat with added nitrate plus vegetables

Eligibility Criteria

• Age: 18 Years - 55 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)

You may qualify if:

• 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.

You may not qualify if:

• 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.

Sponsors & Collaborators

• Edith Cowan University: lead
• The University of Western Australia: collaborator
• Flinders University: collaborator

Study Sites (1)

Royal Perth Hospital Research Foundation

Perth, Western Australia, 6000, Australia

Location

Related Publications (18)

• 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.

  PMID: 20494108 RESULT

• 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.

  PMID: 29635489 RESULT

• 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.

  PMID: 28802834 RESULT

• 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.

  PMID: 1017769 RESULT

• 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.

  PMID: 1017770 RESULT

• 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.

  PMID: 7867685 RESULT

• 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.

  PMID: 7600541 RESULT

• 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.

  PMID: 21141240 RESULT

• 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.

  PMID: 19439460 RESULT

• 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.

  PMID: 28105786 RESULT

• 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.

  PMID: 3057363 RESULT

• 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.

  PMID: 11038239 RESULT

• 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.

  PMID: 6541617 RESULT

• 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.

  PMID: 22336776 RESULT

• 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.

  PMID: 24676365 RESULT

• 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.

  PMID: 27214047 RESULT

• 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.

  PMID: 24288368 RESULT

• 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.

  PMID: 30148503 RESULT

MeSH Terms

Conditions

Health Risk Behaviors

Interventions

Meat; Vegetables

Condition Hierarchy (Ancestors)

Health Behavior; Behavior

Intervention Hierarchy (Ancestors)

Food; Diet, Food, and Nutrition; Physiological Phenomena; Food and Beverages

Study Officials

• Catherine P Bondonno, PhD, RNutr.

  Edith Cowan University

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: SINGLE
• Who Masked: OUTCOMES ASSESSOR
• Masking Details: 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.
• Purpose: PREVENTION
• Intervention Model: CROSSOVER
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: prospective, mono-centre, randomised, controlled, crossover study

Study Record Dates

• First Submitted: August 30, 2021
• First Posted: September 16, 2021
• Study Start: October 11, 2021
• Primary Completion: December 31, 2021
• Study Completion: December 31, 2021
• Last Updated: April 1, 2022

Record last verified: 2022-03

Data Sharing

• IPD Sharing: Will not share

Locations

AU (1)