Presented by: Angela Mastaloudis, Ph.D. Pharmanex Research Institute Provo, UT, USA
An important group of phytonutrients Abundant in fruits and vegetables Epidemiological and clinical studies indicate protection against the development of a variety of chronic diseases - Lutein and zeaxanthin
- -carotene and lycopene
- protection from sun damage
- Lutein and lycopene
- Lycopene
Carotenoids
Relevance of Accurate Assessment of Human Carotenoid Status Marker of oxidative stress - smoking, sunlight exposure, pollution all influence skin carotenoids independent of dietary intake
Survey fruit and vegetable consumption Monitor compliance to diets rich in fruits and vegetables Monitor intake of carotenoid containing dietary supplements
Resonant Raman Scattering of Carotenoids
Resonant Raman Scattering of Carotenoids
Early Studies: The Eye & AMD
BioPhotonic Scanner BioPhotonic Scanner 473 nm excitation, 511 nm detection (C=C)
Skin Carotenoids Carotenoids are measured in the stratum corneum layer of the skin Carotenoids protect epidermis from UV Light Site of action advantage
Skin Carotenoids An Important Indicator of the Body’s Antioxidant Defense System Most convenient measure of antioxidant status Carotenoids measured: - -Carotene, -Carotene, Lycopene, Lutein, Zeaxanthin, -Cryptoxanthin
- First line of defense & part of antioxidant network
Skin carotenoids are influenced by oxidative stress (smoking, sunlight exposure, pollution) independent of dietary intake
Advantages Non-invasive Safe Instant results Inexpensive Highly specific Reflects long-term carotenoid status
Clinical validation of a non-invasive, Raman spectroscopic method to assess carotenoid nutritional status in humans Jeffrey A. Zidichouski1,3, Angela Mastaloudis1, Stephen J. Poole1, James C. Reading2 and Carsten R. Smidt1 1Pharmanex Research Institute, Provo, Utah, USA; 2U. of Utah School of Medicine, SLC, Utah, USA; 3Institute for Nutrisciences and Health, NRC, Canada
Background Serum/plasma carotenoids (HPLC) - currently accepted “gold standard”
- used to validate subjective fruit & vegetable intake data (epidemiological research)
- used to assess human carotenoid and antioxidant status
- invasive & labor-intensive
- not suitable for large populations
Objectives To validate the Raman spectroscopy (RS) methodology (BioPhotonic Scanner) by comparing it to HPLC, the currently accepted gold standard for assessing human carotenoid status using Criterion Validity To assess and compare reliability of the two measures
Methodology 372 adult non-smokers 3 measurements within 8 days, ≥ 48 h apart, after overnight (12 h) fast: - Serum carotenoids by HPLC
- Skin carotenoids, Raman spectroscopy
- Demographic data and food frequency questionnaires
Results
Skin-Serum Correlation
Conclusions Raman spectroscopy (BioPhotonic Scanner) is a valid measure to assess skin carotenoid status in situ in humans RS accurately predicts blood serum total carotenoids, independent of demographic and lifestyle factors Skin carotenoids are more stable over time than serum carotenoids, likely because they are not confounded by recent dietary intake (Dueker SR et al. J Lipid Res. 2000;41:1790) RS appears to be a better indicator of carotenoid status than blood carotenoids
Epidemiological Studies
Fruit & Vegetable Intake
Supplementation: LP* *Multivitamin/mineral Supplement with Antioxidant Nutrients
Body Mass Index
Smoking (Oxidative Stress)
Skin carotenoids: - Are not influenced by age, gender or race
- Increase linearly in response to both dietary intake and intake from supplements
- Are inversely correlated with oxidative stress markers
- (smoking, UV light exposure, urinary F2-Isoprostanes)
- Are a valid indicator of overall antioxidant status
Randomized, Placebo Controlled Antioxidant Supplementation Study
Study Design N = 52 RCT: Twice a day, with breakfast and with dinner, subjects took LP nano or Placebo Study duration: 18 weeks BioPhotonic Scanner - Subjects were scanned a total of 15 times
Lifestyle
Summary Skin carotenoids increased significantly with supplementation ( two weeks) Skin carotenoids increased in all 20 subjects in twenty weeks with supplementation Even after 18 weeks, skin carotenoids were still increasing with supplementation
Double-Blind, Placebo-Controlled Antioxidant Supplementation Study RLI, Florida Hospital Celebration Health 53 subjects who had not taken antioxidant supplements within the last 3 months 6 weeks on supplements (LP or Placebo) Measured skin RS response and serum antioxidants at days 0, 21 and 42
Double-Blind, Placebo-Controlled Antioxidant Supplementation Study
Results: Serum Carotenoids
Results: Skin Carotenoids
Results: Serum Vitamins C and E
Can Skin Carotenoids Serve as a Marker of Antioxidant Status or Oxidative Stress?
The Antioxidant Network
Associations of Antioxidant Status and Oxidative Stress with Skin Carotenoids Assessed by Raman Spectroscopy Joseph Carlson1,3, Shayn Stavens1, Richard Holubkav1, Jeffrey Zidichouski2, Angela Mastaloudis2, Carsten Smidt2, Eldon Askew1 1U. of Utah Salt Lake City, Utah; 2Pharmanex Research Institute Provo, UT; 3Michigan State U. East Lansing, MI
Objectives To evaluate the relationship between skin carotenoids and serum antioxidants (vitamins E and C) To evaluate the relationship between carotenoid antioxidant status and markers of oxidative stress
Study Design N = 307 adult non-smokers Serum carotenoids, vitamin E and vitamin C by HPLC Skin carotenoids using Raman spectroscopy Questionnaires: demographic, lifestyle and dietary data Urinary F2-Isoprostanes
F2-Isoprostanes Gold standard for measurement of in vivo lipid peroxidation Reliable Chemically stable Specific end-product of the free radical mediated oxidation of arachidonic acid (20:4) Possess biological pro-atherogenic activity Plasma concentrations are elevated in subjects with known oxidative stress (smokers, diabetics, obesity, cardiovascular disease, Alzheimer’s Disease etc)
Skin and Serum Carotenoids are Strongly Correlated
Skin Carotenoids are Positively Correlated with Plasma Antioxidants
Skin Carotenoids are Inversely Correlated with F2-Isoprostanes
Summary Skin carotenoids assessed by the Biophotonic Scanner were directly correlated with: - Serum Carotenoids
- Plasma antioxidants (Vitamins E and C)
- Fruit and vegetable intake
Skin carotenoids were inversely related to F2-Isoprosanes, a marker of oxidative stress
Taiwan Data (n = 38,843; April 2006)
Taiwan Data (n = 38,942; April 2006)
Taiwan Data (n = 38,907; April 2006)
Conclusions Measurement of skin carotenoids using Raman spectroscopy (BioPhotonic Scanner) is: - highly correlated with serum carotenoids
- less variable than serum carotenoids
- preferable to serum carotenoids as a marker of fruit and vegetable intake
- Reflective of long-term carotenoid status
- indicative of other serum antioxidants (vitamins C and E)
- indicative of oxidative stress (MDA,F2-Isoprostanes)
Summary Resonance Raman scattering is a viable optical technique to measure skin carotenoids in vivo. It is highly specific, non-invasive and suitable for clinical field measurements of large populations.
Acknowledgments Stephen Poole, Carsten Smidt – Pharmanex Research Institute Jason Morrow – Vanderbilt University Neal Craft – Craft Technologies, NC Lester Packer – University of Southern California Kyung-Jin Yeum, Jeff Blumberg – Tufts University James Rippe – Rippe Lifestyle Institute Wayne Askew, Joe Carlson, Shayn Stavens - University of Utah, Division of Foods and Nutrition, SLC
Jeff Zidichouski – Canadian National Research Council (P.E.I.) Werner Gellermann – University of Utah, Physics Dept., SLC
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