Spice as Medicine, Mustard seeds, Brassica nigra, Brassica juncea, Sinapis alba, nutrient composition, bioactive compounds, glucosinolate, sinigrin, sinalbin, myrosinase, isothiocyanate, health benefits, toxicity, mustard green, mustard, oil, mustard essential oil

1. SPICE AS MEDICINE:
MUSTARD
GENUS BRASSICA &
GENUS SINAPIS
By
Kevin KF Ng, MD, PhD
Former Associate Professor of Medicine
Division of Clinical Pharmacology
University of Miami, Miami, CL, USA
Email: kevinng68@gmail.com
A Slide Presentation for HealthCare Provider Seminar July 2019

2. Outline of Lecture
▪ Historical milestones..
▪ Scientific classification, production & consumption of mustard seed.
▪ Anatomy of mustard seed, nutrient composition of mustard seed.
▪ Bioactive compounds of mustard seed.
▪ Chemistry, bioavailability.
▪ Health benefits.
▪ Clinical studies and meta-analysis.
▪ Commercial mustard seed products.
▪ Summary.

3. Historical milestones of Mustard

4. What is Mustard ?
Ground
mustard
seed
Liquid
(vinegar)
etc.
Mustard

5. Varieties of mustard seeds
Mustard is a condiment made from the seeds of a
mustard plant. There are three varieties:
• White/yellow mustard, Sinapis alba
• Brown/Indian mustard, Brassica juncea
• Black mustard, Brassica nigra

6. Difference between mayonnaise and mustard
▪ Mayonnaise is a mix of raw eggs, oil, and spices.
▪ A serving of mayonnaise is one tablespoon and has 90
calories, 6 times the amount in ketchup.
▪ A serving of mustard is one teaspoon and has only 5
calories, no sugar, no fat and only 55mg of sodium. The
flavor is stronger than mayo or ketchup.
mayonnaise

7. Scientific classification of Mustard plant

8. World’s Top Mustard Seed Producing Countries
https://www.worldatlas.com/articles/the-world-s-top-mustard-seed-producing-countries.html

9. Top Mustard Seed Consuming Countries in the World
1. Nepal
2. Canada
3. Russia
4. Ukraine
5. Myanmar
https://www.indexbox.io/blog/which-country-eats-the-most-mustard-seeds-in-the-
world/

10. Three principal mustard species

11. Anatomy of a mustard tree and seed
▪ Size of bush/tree: 6-20 ft
▪ Size of seed: 1-2 mm diameter
mustard tree mustard seedmustard seed Magnified mustard seed

12. Comparison of black and white mustard seeds
Adapted from https://www.slideshare.net/MarwaFayed1/seeds-52154912

13. Nutrient composition of ground mustard seed
Source: USDA National Nutrient Database for Standard Reference 1 April 2018 Software v.3.9.5.3_2019-06-13

14. Fatty acids and Bioactive compound in ground mustard seed

15. Summary of nutrient composition of
Mustard Seeds
Sinigrin
Sinalbin
33%

16. Scientific studies on mustard

17. Number of articles on Mustard, Glucosinolate and Sinigrin in
PubMed from 1918 to 2018
https://www.ncbi.nlm.nih.gov/pubmed/?term=sinigrin

18. Formation of pungent compound from glucosinolate
(pungent)
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6273501/figure/molecules-21-00416-f002/

19. Chemical reaction of mustard glucosinolates
from yellow and brown/black mustard
Chemical reaction of mustard glucosinolate:
(a) Sinalbin in the presence of water reacts
with myrosinase to form p-hydroxybenzyl
isothiocyanate (sharp taste without
pungent aroma).
(b) Sinigrin in the presence of water reacts
with myrosinase to form allyl
isothiocyanate (pungent irritating
odour).
(Source: Tainter and Grenis 2001.)
https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/sinigrin

20. Effect of heat on relative myrosinase activity in
black, brown and yellow mustard seeds;
where temperature exposure time was 10 min.
https://docksci.com/thermal-and-pressure-stability-of-myrosinase-enzymes-from-black-mustard-
brassica_5a4a380bd64ab206abf27c44.html

21. Flavors of white, brown and black mustards

22. Enzymatic hydrolysis reaction of glucosinolates and their
breakdown products.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4985713/

23. Fate of glucosinolates and their breakdown products in the
human gut.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4985713/

24. Pharmacology of Bioactive compounds of
Mustard Seeds

25. Effect of glucosinolate and its degradation products
on health
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4031110/
Anticancer
Flavor
Reduce
thyroxine
Reduce
thyroxineToxic?

26. In vitro anti-hyperglycemic action by sinigrin (2017)
https://www.sciencedirect.com/science/article/pii/S1995764516307611
sinigrin (enzyme inhibitor)

27. Sinigrin inhibits production of inflammatory mediators by suppressing
NF-κB/MAPK pathways or NLRP3 inflammasome activation in
macrophages. 2017
▪ Sinigrin (2-propenyl glucosinolate) is found mainly in broccoli, brussels sprouts, and black mustard seeds.
▪ This study investigated the effect of sinigrin on macrophage function, including the activity of Nod-like receptor
protein 3 (NLRP3) inflammasome.
▪ In a concentration-dependent manner, sinigrin inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO)
production and the expression of COX-2 and prostaglandin E2 (PGE2) in RAW 264.7 cells.
▪ Sinigrin suppressed the production of tumor necrosis factor (TNF)-α and interleukin (IL)-6 via suppression of MAPK
phosphorylation and nuclear factor-kappa B (NF-κB) activity.
▪ Treatment with sinigrin decreased IL-1β and IL-18 production and concurrently suppressed NLRP3, apoptosis-
associated speck-like protein containing a caspase recruitment domain (ASC), and caspase-1 expression in
LPS/ATP-stimulated cells, suggesting that the blocking of NLRP3 inflammasome activation prevented the
production of both cytokines.
▪ These results suggest that sinigrin has immunomodulatory effects by suppressing the production of inflammatory
mediators, possibly by inhibiting the NF-κB/MAPK pathways or NLRP3 inflammasome activation. Our findings also
provide evidence that the pharmacological modulation of sinigrin could have an anti-inflammatory effect.
https://www.ncbi.nlm.nih.gov/pubmed/28219839

28. Isothiocyanates (ITCs) inhibit NF-kB –mediated inflammation
(B) and induce nuclear translocation of Nrf2 (A)
https://lpi.oregonstate.edu/mic/dietary-factors/phytochemicals/isothiocyanates
ITCs induce nuclear translocation of Nrf2 ITCs induce nuclear translocation of Nrf2

29. Sinigrin attenuates the progression of atherosclerosis in ApoE−/− mice
fed a high-cholesterol diet via reduction of proinflammatory
cytokines and chemokines
https://www.sciencedirect.com/science/article/pii/S0009279717302144
VCAM-1 ICAM-1
CCL2 CCL5
VCAM-1=Vascular Cell Adhesion Molecule 1
ICAM-1=Intercellular adhesion molecule-1
CCL2=Chemokine ligand 2
CCL5=Chemokine ligand 5

30. Anti-cancer Activities of Sinigrin on Carcinogen-Induced
Hepatotoxicity in Rats (2014)
▪ Sinigrin is one of the major ingredients present in Brassica nigra, which has been used in combination with other herbs
for treatment of various diseases.
▪ The anti-proliferative activities of sinigrin were studied in a model of carcinogen-induced hepatotoxicity in rats.
▪ Rats were orally administered with sinigrin on a daily basis for three months before sacrifice.
▪ Sinigrin was found to significantly inhibit the proliferation of liver tumor cells; the number of surface tumors in the rat
liver was dramatically reduced.
▪ Sinigrin induced apoptosis of liver cancer cells through up-regulation of p53 and down-regulation of Bcl-2 family
members and caspases.
▪ Our findings indicated that the liver functions were gradually restored after treatment with sinigrin and that the agent did
not cause liver toxicity.
▪ Cell cycle analysis indicated that sinigrin caused cell cycle arrest in G0/G1 phase. The results suggest that sinigrin
exerts important anti-proliferative activities in carcinogen-induced hepatocarcinogenesis in rats, and highlight the
potential of sinigrin as an anti-cancer agent for liver cancer.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4203766/

31. Anticancer activity of Sinigrin and Sinigrin plus Myrosinase on
cancer cells in vitro. (2015)
Adapted from https://www.mdpi.com/1420-3049/20/9/15748

32. Myrosinase in Mustard Seeds Increase the Bioavailability of Sulforaphane in
Healthy Human Subjects after the Consumption of Cooked Broccoli.
https://www.ncbi.nlm.nih.gov/pubmed/29806738

33. Some biological activities of selected glucosinolate
breakdown products (ITCs)
Adapted from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4985713/

34. Uses of mustard plant: leaf, seed, mustard, oil

35. Varieties of Mustard
▪ Yellow mustards: American mustard
▪ Dijon : classic French mustard
▪ Spicy Brown: coarser grind
▪ Wine mustards: white burgundy, pinot noir, champagne
▪ Coarse mustard: creole, stone ground, whole grain, Dijon, whole grain stout, whole
grain whisky
▪ International mustards: English, German, Chinese
▪ Sweet mustards: honey mustard, pecan-honey, sweet-hot, brown sugar, honey Dijon
▪ Flavored mustards: horseradish, sriracha, balsamic, habanero
https://www.foodrepublic.com/2016/03/10/a-guide-to-all-the-different-kinds-of-mustard/

36. What is mustard oil?
https://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-mustard-essential-oil.html

37. Fatty acid composition of vegetable and mustard oils
Adapted from https://slideplayer.com/slide/14689533/

38. Fatty acid composition of mustard oil
▪ Mustard oil has about 60% monounsaturated fatty acids:
▪ 42% erucic acid (omega 9 fatty acid)
▪ 12% oleic acid
▪ About 21% polyunsaturated fats:
▪ 6% the omega-3 alpha-linolenic acid
▪ 15% the omega-6 linoleic acid
▪ About 12% saturated fats. erucic acid

39. What is Mustard Essential Oil?
▪ Mustard essential oil, often mistaken for mustard oil, is
made from mustard seeds through a distillation process.
▪ Mustard essential oil is also called volatile oil of mustard.
▪ The essential oil contains 92% allyl isothiocyanate, which
is the compound responsible for the pungent taste of
mustard.
▪ It is this allyl isothiocyanate, along with important fatty
acids like oleic acid, linoleic acid, and erucic acid, which
contribute to the long list of medicinal benefits of mustard
essential oil.
▪ While it is safe to consume in small amounts, the essential
oil is usually used topically.
https://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-mustard-essential-oil.html

40. Fatty acid composition of mustard essential oil
Adapted from https://vdocuments.mx/physico-chemical-properties-of-mustard-oil-bio-medical-application-of-ultrasonic.html
Mustard essential oil
distillation
Toxic

41. GLC analysis of fatty acids in vegetable and Mustard oil
https://vdocuments.mx/physico-chemical-properties-of-mustard-oil-bio-medical-application-of-ultrasonic.html

42. Toxicity of Erucic acid in mustard oil
▪ Erucic acid is a monounsaturated omega-9 fatty
acid, denoted 22:1ω9.
▪ Research showed that long term usage of erucic
acid in animals caused myocardial lipidosis
▪ Mustard is not safe for use as a cooking oil.
▪ Mustard oil is banned in US, Canada and EU

43. Ingredients in modern day mustard
Mustard seed
White
Brown
Black
Liquid
Grape juice
Vinegar
Wine
Syrup
Water
Ingredient
Salt
Garlic
Chili Pepper
Spice
Tumeric

44. Safety issues
▪ Adverse effects: no serious effects of glucosinolates and
isothiocyanates in humans have been reported
▪ Thiocyanate generated from consumption of indole
glucosinolate can be assumed to have minimal adverse
risks for thyroid health.
▪ Drug interaction: glucosinolates and isothiocynates are
not known to interact with any drugs.
▪ The statement: “For External Use Only” indicates that
the mustard oil should not be used as a food ingredient.
▪ EU Directive 80/891/EEC requires that erucic acid
content of foods to be no greater than 5%

45. Examples of some delicious mustard cuisine

46. Summary
▪ Mustard is a paste made by mixing ground mustard seed with a liquid.
▪ The mustard seed is derived from three varieties of mustard plants.
▪ Present in the mustard seed are macronutrients, micronutrients and
bioactive glucosinolates.
▪ The enzyme myrosinase acts on glucosinolate to produce isothiocyanate
and other products which are responsible for the pungent flavor,
antimicrobial, anti-inflammatory, immuno-modulatory and anti-cancer
properties.
▪ The high content of erucic acid in mustard oil makes it unsafe to be used
as a cooking oil.
▪ Mustard is safe for ordinary consumption as a spice. No serious adverse
reaction has been reported

47. ANY QUESTION?
Please contact
Kevin KF Ng, MD, PhD
Email: kevinng68@gmail.com