A 12-week, parallel-group, two-arm, single-center, randomized, double-blind, placebo-controlled trial was completed. The trial protocol was approved by the Allendale Institutional Review Board, and this study conformed to the standards set by the latest revision of the Declaration of Helsinki. This study was retrospectively registered at clinicaltrials.gov (ClinicalTrials.gov trial registration NCT06749808) in December 2024. The study was conducted between July and October 2024.

Forty-eight healthy males and females between the ages of 20 - 65, BMI ≥ 18.5 and ≤29.9 kg/m², with a score of ≤30,000 Raman intensity counts (RIUs) using Raman spectroscopy (BioPhotonic Scanner S3, Pharmanex) were recruited for this study. Subjects were randomly assigned to either the supplement group (SUP) (n = 29) or the placebo group (PL) (n = 17). Unequal group allocation was planned a priori. For the study endpoints, the precision of the treatment effect estimate depended more heavily on the size of the active group than the placebo group. The small loss of power was adjusted by increasing the total sample size. Additionally, as biomarker analysis was a key part of the study objectives, a larger active group was planned to increase the informative value of these analyses. The placebo served as a comparator, not the main object of the study. Each group was instructed to take the supplement or placebo twice daily (morning and evening with meals) for 12 weeks. They were also instructed not to change their eating or exercise habits for the duration of the study.

Subjects completed in-person visits to the research facility at baseline and weeks 1, 2, 4, 8, and 12. At each time point, subjects were given questionnaires on quality of life (QOL), skin attribute assessment, and completed non-invasive carotenoid skin measurements. Resonance Raman Spectroscopy (RRS) using a BioPhotonic Scanner S3 (Pharmanex, UT) was compared to a custom-built hyperspectral absorption device (HA) being developed (Rhyz Analytics, UT) that uses Reflection Spectroscopy. Hyperspectral absorption spectroscopy was utilized in the custom-built device. The device sequentially illuminated the fingertip with four discrete color bands: cyan, red, yellow, and green. A total of 798,720 optical measurements were acquired during each 15-second measurement. Incident and diffusely reflected light were recorded for each illumination cycle using a dual-photodiode configuration. One photodiode continuously monitors LED radiant output for real-time source normalization, while a second photodiode measures reflected light intensity following photon-tissue interaction and re-entry into the device. Wavelength-dependent absorption was quantified as a decrease in reflected intensity relative to the monitored incident illumination. Subjects also filled out a diet and lifestyle questionnaire at baseline and week 12. Additionally, at baseline and week 12, all subjects had blood draws completed to analyze for Vitamin C and Selenium levels. All subjects also had blood draws at baseline and week 12 for a comprehensive metabolic panel.

Study volunteers were recruited through email databases in April 2024. Eligible participants were randomly assigned to one of two groups (SUP or PL). A participant identification number was allocated to each participant based on the order of enrollment. Identification of test articles was stored by the study sponsor and revealed after all data were analyzed. All test articles were packaged in identical packaging. Study investigators were blind to the treatment assignment until all study outcomes were collected.

Forty-eight (48) subjects were enrolled in the study. Forty-six (46) subjects completed all aspects of the study. Subjects satisfied the inclusion and exclusion criteria, accepted the prohibition and restrictions, and gave informed consent. The suitability of each subject to participate was confirmed prior to their acceptance into the study by completion and review of a pre-study questionnaire to determine eligibility. The inclusion criteria comprised healthy males and females between ages 20 - 65 years, BMI ≥ 18.5 and ≤29.9 kg/m², with a score of ≤30,000 Raman intensity counts (RIUs) using Raman spectroscopy (BioPhotonic Scanner, Pharmanex), no plans to start new treatments during the study, able to understand the study procedures, and willing and able to adhere to all study procedures. The exclusion criteria comprised pregnant women, women who were nursing, or women planning to become pregnant; subjects who were currently taking vitamin C, selenium and carotenoid-containing supplements; known allergies to ingredients in the study dietary supplement; subjects with history of any gastrointestinal surgery and/or illness which in the opinion of the investigator would affect the digestive and absorption process of the body; use an indoor tanning booth; cigarette smokers, alcohol or narcotic addicts; concurrent therapy with any medication that might interfere with the study.

The supplement was provided as four capsules in a sachet taken twice daily (8 capsules daily) containing 3 mg boron, 255 mg calcium, 100 mcg chromium, 0.9 mg copper, 150 mcg iodine, 180 mg magnesium, 75 mcg molybdenum, 2.3 mg manganese, 140 mcg selenium, 20 mcg vanadium, 15 mg zinc, 55 mg choline, 7.5 mg beta-carotene, 150 mcg biotin, 30 mg pantothenic acid, 122 mcg methyltetrahydrofolic acid, 118 mcg folic acid, 32 mg niacinamide, 10 mg pyridoxine, 8.5 mg riboflavin, 7.5 mg thiamine, 375 mcg vitamin A, 16 mcg methylcobalamin, 14 mcg vitamin cyanocobalamin, 1000 IU vitamin D3, 30 mg vitamin E, 40 mcg vitamin K2, 400 mg vitamin C, 150 mg grape seed extract, 40 mg olive leaf extract, 100 mg alpha lipoic acid, 2 mg lutein, 5 mg lycopene, 30 mg quercetin, and 5 mg resveratrol.

The placebo was provided as four capsules in a sachet taken twice daily (8 capsules daily) containing maltodextrin.

Each group was instructed to take the supplement or placebo twice daily with a meal (morning and evening) for the 12-week trial period. Subjects were also instructed not to change their eating or exercise habits for the duration of the study. Daily diaries were administered to assess product compliance.

In-person assessments were conducted at Weeks 0 (baseline), 1, 4, 8, and 12. Self-report diaries were also completed daily during the 12-week study duration. Primary outcome measures included at baseline, weeks 1, 4, 8, and 12 were skin carotenoid concentration measured on the finger via Hyperspectral Absorption (HA) with a custom device being developed, and on the palm using Resonance Raman Spectroscopy (RRS) using the BioPhotonic Scanner S3. Secondary outcome measures included at baseline and end of study (week 12) were changes in feelings of wellness as measured by the Quality of Life (QOL) questionnaire, skin appearance attribute changes via self-assessment, serum vitamin C levels, and serum selenium levels. Secondary outcome measures for safety included a comprehensive metabolic panel via blood draw at baseline and week 12, and self-reported adverse events (AEs) were assessed at each study visit to examine the safety and tolerability of the test articles.

Descriptive statistics (mean and standard deviation) and change from baseline (T-Test, percent of subjects improving) were conducted for all study outcome evaluations. A Welch’s T-Test was used, assuming unequal variances, and to account for differences in sample size for each group. For the Self-Perception questionnaires, frequency of responses was summarized for each question. Within-treatment analyses were done using a Binomial test with a null hypothesis of a 50/50 split of top box/not top box responses. A Mann-Whitney U Test was conducted for the categorical data responses. All statistical tests of hypotheses employed a level of significance of 0.05.

Forty-eight (48) subjects were enrolled in the study. Twenty-nine (29) subjects enrolled in the dietary supplement group completed the study, while seventeen (17) subjects enrolled in the placebo group completed the study. Two subjects enrolled in the dietary supplement group were lost due to scheduling conflicts. Baseline demographic characteristics are detailed in Table 1.

Table 1.Subject demographic characteristics.

Skin Carotenoid Concentration: As detailed inTable 2, subjects in the SUP group had statistically significant improvement in BioPhotonic Scanner (RRS) scores over baseline and compared to placebo at weeks 4, 8, and 12. The PL group did not have a significant change from baseline throughout the study. By week 12, the SUP group had a 42% increase from baseline. As detailed in Table 3, subjects in the SUP group had statistically significant improvement in hyperspectral absorption device (HA) scores at weeks 8 and 12. The PL group did not have a significant change from baseline throughout the study. By week 12, the SUP group had a 41% increase from baseline score.

Table 2.BioPhotonic S3 scanner results (RRS).

a. p < 0.05.

Table 3.Hyperspectral absorption device results (HA).

a. p < 0.05.

Nutrient Status: Serum vitamin C levels significantly increased from baseline to the end of the study at 12 weeks in the SUP group compared to the placebo (p = 0.0004) (Table 4). The SUP group had a 44% increase in vitamin C levels from baseline. Serum selenium levels also significantly increased from baseline to the end of the study at 12 weeks in the SUP group compared to placebo (p = 0.000001) (Table 5). The SUP group had a 26% increase in selenium levels from baseline. No changes were seen for vitamin C and selenium levels in the PL group.

Table 4.Serum vitamin C levels.

a. p < 0.05.

Table 5.Serum selenium levels.

a. p < 0.05.

Figure 1. Quality of life results.

Quality of Life (QOL): At Week 12, participants were given a questionnaire that asked questions about quality-of-life parameters. The participants were asked to rate their experience on a 7-point scale, where 1 is strongly disagree, and 7 is strongly agree. A positive response was defined as any level of agreement to the question (including ratings of 5, 6, or 7). Results from the QOL questionnaire indicate subjects in the SUP group perceived significantly greater benefit across all QOL attributes assessed, when compared to the PL group (Figure 1).

Skin Appearance Attributes: The subjects were asked to assess questions about skin attributes using the same 7-point scale. Results indicate subjects in the SUP group perceived significantly greater benefit across all skin attributes assessed, when compared to PL. Summary of results presented in Figure 2.

Figure 2.Skin appearance attributes.

Safety and Tolerability: No adverse events were reported during the study, indicating the dietary supplement was well tolerated. Additionally, no meaningful changes were seen with SUP compared to PL or baseline in the comprehensive metabolic panel (CMP) used for safety.