Whats up doc? Bugs Bunny asked, crunching on a renowned beta-carotene source. The simple answer? Carotenoids are whats up. Both on screen and off they appear in Technicolor. They are found in a cornucopia of vegetables and fruitspeaches to peppers, cantaloupe to kale and, of course, carrots. Beta-carotene is one of 600 types of carotenoids. Other major ones seen in the dietary supplements and functional foods industry are lycopene, astaxanthin, lutein, zeaxanthin and alpha-carotene.

Carotenoids protect cells from damage caused by unstable free radicals, which are believed to contribute to certain chronic diseases and play a role in the degenerative processes seen in aging. Their antioxidant properties plus added health benefits are driving market demand.

However, the rising star in the field is lutein. Consumers have heard the scientific message that lutein may help reduce age-related macular degeneration (AMD), pushing luteins market value up to $139 million in 2004, according to BCC Research. While no recommended daily allowance (RDA) currently exists for lutein, as it does for other nutrients, the only definitive side effect of excess lutein consumption is the same observed for beta-caroteneoverdose can result in tanning of the skin.

Beta-carotene is used in the dietary supplements area primarily as a source of vitamin A and in the food industry as a colorant along with apocarotenal and canthaxanthin. Supplements based on free lutein, lutein esters, astaxanthin and zeaxanthin are available in the form of tablets, soft-gelatin and two-piece liquid capsules, hard gelatin capsules, dry mixes, beverages and functional foods.They are used in nutritional supplements to replenish carotenoids in people leading urban lifestyles, who are typically deficient in plant products.

Carotenoid inclusion in nutritional beverages (powder & RTD) and bars is slowly gaining momentum. Dietary supplement applications have expanded to include effervescents, oral strips, gummies, liquid drops and an assortment of chewable formats including hard candies, said Philip J. Gowaski, sales and marketing manager, Chrysantis. And according to Charles DePrince, president, Fuji Health Science Inc., carotenoids can be used for a variety of functionsmuscle endurance and recovery (reducing the build-up of lactic acid and limiting damage to the muscle cell membrane following strenuous exercise), skin health (reducing the appearance of fine wrinkles, increasing the elasticity and moisture level of skin), asthenopia (reducing eye fatigue), gastric health (reducing inflammatory effect of H. pylori infection and dyspepsia) and other areas of application still under study, such as cardiovascular, diabetes, weight loss and metabolic syndrome.

Targeted Nutrition

Each carotenoid offers specific health benefits, including vitamin A activity and antioxidant properties, anti-cancer properties, eye health support, immune enhancement, inflammation modulation, and much more. As long as the bodys level of vitamin A is adequate, it uses betacarotene as a singlet oxygen quencher; typically, only about one-third of ingested beta-carotene converts to vitamin A.

According to the Journal of the Science of Food and Agriculture (81: 559-568, 2001), there are three main categories of antioxidants: excellent antioxidants that perfectly quench excited states as well as ground state radicals (e.g., astaxanthin); good antioxidants that strongly inhibit peroxide formation but are less efficient in quenching excited states (flavanols, tocopherols)or lead to considerable degradation of the antioxidant itself (beta carotene, lycopene); and moderate antioxidants that fail to excel in both states (alphacarotene, flavone).

Improved general eye health is a well-known benefit from carotenoids. Within the central macula of the eye, zeaxanthin is the dominant component, whereas in the peripheral retina lutein dominates. Therefore, more product developers are using a combination of both lutein and zeaxanthin for eye health formulations. In a study published in Opthalmic and Physiological Optics (26, 4: 362, 2006), 34 subjects received six months of daily supplementation of zeaxanthin (Z, Optisharp, from DSM Nutritional Products), lutein (L), a combination of the two (L-Z) or placebo. Subjects were then given doubled supplementation. Data revealed a trend toward lower CATs in all groups supplemented, with statistical improvement in the lutein group. Supplementation with L or Z increased macular pigment density at the fovea and at 2.5 degrees, and improved CATs at high mesopic levels and, hence, visual performance at low illumination. Further studies have shown an inverse association between high dietary L-Z intake and prevalence of nuclear cataract ( J Invest Ophthlmol Vis Sci, 47, 9:3783-6, 2006). And data from the Carotenoids in Age-Related Eye Disease Study (CAREDS) showed diets rich in lutein plus zeaxanthin may protect against intermediate AMD in women younger than 75 (Arch Opthalmol, 124:1151-62, 2006).

Lycopene, along with alpha- and beta-carotenes, has been associated with cardiovascular health. In one recent Japanese study, high serum levels of alpha- and beta-carotenes and lycopene reduced the risk for mortality from cardiovascular disease (CVD) (J Epidemiol, 16, 4:154-60, 2006). Another trial found the combination of olive oil and lycopene increased serum lycopene levels, increased levels of good high-density lipoprotein (HDL) cholesterol, and lowered levels of triglycerides and bad low-density lipoprotein (LDL) cholesterol (Asia Pac J Clin Nutr,12 Suppl:S21, 2006). While much research has examined the impact of carotenoids on CVD and atherosclerosis, more trials are still ongoing. Researchers from Brigham & Womens Hospital and Harvard Medical School, Boston, noted in a published review that emerging evidence of potential cardioprotective effects for lycopene and other carotenoids (besides beta-carotene) in both the diet and blood suggest that there is more to be learned (Curr Opin Lipidol, 7,1:11-6, 2006).

Another research focus is the ability of carotenoids to protect against oxidative DNA damage that contributes to numerous chronic diseases. Researchers from the U.S. Department of Agriculture (USDA) Jean Mayer Research Center at Tufts University, in conjunction with scientists from the University of Milan, Italy, and BASF investigated the effects of daily supplementation with mixed carotenoids (betacarotene, lutein and lycopene: 4 mg each), 12 mg of a single carotenoid (beta-carotene, lutein or lycopene) or placebo in older women for a total of 56 days (Am J Clin Nutr, 83, 1:163-9, 2006). Each supplemented group exhibited significantly reduced DNA damage compared to baseline, whereas the placebo group did not show the same improvement. The mixed carotenoid and the beta-carotene groups further displayed significantly less oxidative DNA damage as early as 15 days into supplementation.

This ability to protect DNA may be one reason carotenoids appear to prevent cancer progression. Consider, for example, the results of a study out of the All India Institute of Medical Science, New Delhi, in which 54 men with metastatic prostate cancer were randomized to receive orchidectomy (testicle removal) alone or orchidectomy plus Lyc-O-Mato® tomato lycopene complex (from LycoRed) providing 2 mg lycopene plus other tomato phytonutrients, twice daily (BJU International, 92:375-8, 2003). After two years, researchers reported adding tomato lycopene complex to the diet produced a more reliable and consistent decrease in serum prostate-specific antigen (PSA) levels, helped shrink the primary and secondary tumors, provided better relief from bone pain and lower urinary tract symptoms. Another study published in the Journal of Herbal Pharmacotherapy found astaxanthin demonstrated 98 percent inhibition of 5alpha-reductase at 300 micrograms/mL in vitro (5, 1:17-26, 2005). A combination of astaxanthin with saw palmetto berries (as Alphastat, from Triarco) had a 20-percent greater inhibition.

Increased intake of lutein and zeaxanthin may also help in cancer prevention. A case-control study coordinated by the National Cancer Institute (NCI) identified 466 subjects with non Hodgkins lymphoma (NHL) and matched them to controls (n=391) for age, race and sex from four different U.S. cancer research centers (Am J Clin Nutr, 83, 6:1401-10, 2006). NHL risk was inversely associated with higher intake of all vegetables (multivariable odds ratio for highest to lowest quartile, 0.58); with green leafy vegetables (OR 0.62); and with greater daily intakes of lutein and zeaxanthin (OR 0.58) and zinc (OR 0.58).

Making 24 Carrot Choices

While consumers have traditionally turned to dietary supplements for bolstering nutrient intake, they are increasingly expecting added value from their foods and beverages, opening up a new field for manufacturers, said Scott Larkin, marketing manager with LycoRed.

Overall there is a clear trend for food and beverage companies towards making their products healthier, agreed Caroline Brons, senior marketing manager at DSM Nutritional Products. And it is not limited to the United States; the global market is also aware of the applications. John R. Benemann, Ph.D., with Parry Nutraceuticals Ltd., added, There has been a good response from the European food and beverage companies looking for carotenoids with strong antioxidant and color quality that are also a safe source of vitamin A.

Companies are making strides toward broadening product portfolios to be more inclusive. LycoRed, best known for its Lyc-OMato tomato complex recently launched a Lyc-O-Lutein ingredient. And OmniActives is manufacturing both patented free lutein and lutein esters in a patent-pending vegetarian beadlet and oil suspension forms for tablets, capsules and soft-gel application, as well as stable, water-dispersible forms. With the introduction of the new Dietary Guidelines, consumers are becoming more aware of the difficulty in getting enough of these key carotenoids through their diet, said Sanjaya Mariwala, managing director, OmniActive Health Technologies.

We see an increased interest by manufacturers to help their customers fill this nutrient gap, agreed Rob Bailey, marketing manager at Cognis Nutrition & Health.

Volatile, But Valuable

Working with these highly potent antioxidants poses more than a few challenges for formulators, but the resulting benefits are worth it. Even sourcing is a challenge for some carotenoids; consider how astaxanthin, primarily sourced from cultivated algae must be protected from contamination. Active carotenoid ingredients are heat sensitive and susceptible to interaction with other active compounds in formulation. They are lipophilic, which makes them generally insoluble in water. And they are quite prone to oxidative degradation by light or heat and can be chemically unstable in acids.

Gowaski cited stability as the major challenge; but, the compounds are also sensitive to light, pressure, pH, oxygen and temperature. Formulators must make certain the carotenoids they choose can withstand the production process and interaction with other ingredients, he said.

Benemann agreed: The product has to be carefully processed, stabilized and properly stored and handled to ensure that there is no deterioration of the pigments during processing, formulation and shelf life of the product. So getting antioxidants to the consumer can be a tricky and arduous task without losing the important benefits carotenoids have to offer.

Being attentive to the challenges posed in formulation and selecting the appropriate concentration is critical. This means generally starting with the most concentrated beadlets first to keep the dosage size down and during blending, [carotenoids] may be added to the general mix or it can be included in a pre-mix of ingredients, said Nicki Jacobs, vice president of technical and regulatory affairs at B&D Nutritional Ingredients.

Missy Schaaphok, research & development scientist, Seltzer Nutritional Technologies, agreed attention to detail is fundamental in formulation. Be aware of the color effect of carotenoids, she said. Many, such as beta-carotene, exhibit a yellow to orange coloration. And when formulating a liquid, be aware most carotenoids are oil soluble only.

Traditionally, manufacturers have used dry forms standardized in a matrix of animal origin gelatin. As natural isolates, lutein and luteinesters tend to be highly viscous, colored, unstandardized and hence very difficult to work with, agreed Mariwala.

Ensuring the body can absorb the carotenoids is another challenge, noted Robert Berman, senior marketing manager at DSM Nutritional Products. In addition to other factors, bioavailability is dependent on the particle size of the carotenoid in the inner phase of the beadlet or the micronized crystals in oil suspensions, he said. The smaller the particle size, the better the bioavailability.

Jacobs also raised the issue that carotenoid manufacturers may face negative media or a past study. Probably the single biggest challenge is overcoming stigma associated with the outcomes of the CARET study where long term high dosages of betacarotene were implicated as being potentially problematic for heavy smokers, she said. Like many, she believes the benefits do far outweigh any risk of toxicity in retinol, retinol acetate, or retinol palmitate.

Taste can be another issue formulators contend with in developing a marketable product depending on the carotenoid and what format (powder, oil or beadlet) is used for the formula. If it tastes bad, people will unlikely want to consume it. Other sensory challenges are smell and color; both distinguishable and potent characteristics of carotenoids.

Gowaski explained, Most carotenoids are strong colorants that will affect the pigmentation of the finished product requiring formulators to limit amounts, use encapsulated products or adjust the original color.

However, knowledge of the issues can help ensure they are appropriately addressed in the development stage, Bailey said. Regulation and a technical group with expertise managing sensory issues are important to successful product launches.

Stable Choices

Quality control and stability begins at the seed production level and validation and verification of stability, concentration and microbial contamination is monitored at every step until the product is sold as a finished good.Manufacturers have formulated a variety of solutions; carotenoid products for consumer health come in various forms, depending on the productincluding encapsulated into beadlets or provided as an oil suspension making it more bioavailable.

Extensive laboratory analysis must be done to ensure safety, purity and [carotenoid] content by approved analytical methods, said Bob Capelli, vice president sales with Cyanotech.

Schaaphok said there are methods to ensure high quality. For example, she said, We conduct identity and potency testing, and proper storage in a cool dry environment is important to maintaining a longer shelf life.

With the sale of carotenoids on the rise, technology is finding more ways to bring health to consumers. Brons said DSM offers a patented beadlet technology (Actilease) designed to significantly increase stability by using an actilease beadlet containing nanodroplets of micronutrients in a protective (water soluble) food starch matrix; preventing losses during manufacturing. One of Fujis basic technologies is the powdering of oils. This is done in-house using proprietary systems. Our powdered astaxanthin AstaReal P2 AF is stable and formulated as vegetarian without the use of allergens and is also cold water dispersible, said DePrince.

Looking for The Real Thing

The difference between synthetic carotenoids and those found in nature can be profound, down to molecular level. The human body utilizes natural and synthetic betacarotenes entirely differently, for example. Natural carotenoids come complexed with fatty acids attached to the end of the molecule making it esterified, Capelli said. However, he noted the greatest difference is in how synthetic works versus natural. In one test of antioxidant strength (free radical elimination) at Creighton University, BioAstin natural astaxanthin was shown to be 20 times more powerful as an antioxidant than synthetic, he said. Were really talking about two completely different products.

Synthetic beta-carotene goes directly to the liver and is stored as retinol palmitate; whereas natural source beta-carotene is absorbed but only what the body needs is sent to the liver to be stored as retinol palmitate. This is important for the elderly population because, as Jacobs explains, Seniors at risk for hip fracture should be cautious with their vitamin A intake, so allowing the body to self-regulate its uptake by consuming [natural] beta-carotene makes a lot of sense. There are also issues with the cisand trans-isomers of beta-carotene being of varied utility to the human body. She adds, beta-carotene derived from Blakeslea trispora offers the highest percentage of the desirable trans- isomer.

Cis

- and trans-isomers and the ratios of the isomers vary in nature depending on the source and processing. Synthetic carotenoids also have varying ratios of isomers but these are usually are in the same range as natural products. The synthetic carotenoids have a chemical composition identical to nature. Synthetic carotenoids also have varying ratios of isomers but these usually are in the same range as in the natural products, said James Elliott, Ph.D. scientific leader, DSM.