Novel ELISA technology in assessing undenatured type II collagen in functional foods and dietary supplements used for knee joint health care: its sensitivity, precision, and accuracy
Introduction: Undenatured type II collagen, derived from chicken sternum cartilage, is a novel functional ingredient, which has been demonstrated to improve joint health, flexibility and mobility, and enhancing motor functions. Undenatured type II collagen has been commercially available as functional dietary supplement worldwide for many years. Research studies demonstrated its broad-spectrum safety and clinical efficacy. Undenatured type II collagen requires very small amount to exhibit clinical efficacy and hence can be easily consumed over a long period of time as compared to the other joint care functional ingredients such as glucosamine and chondroitin. Since undenatured type II collagen is effective in relatively small amount, its accurate measurement in various dosage forms such as tablets and capsules become crucial to provide consumers optimal cost and joint-health benefits.
Objective: In the present study, we modified the previously used Enzyme-Linked Immunosorbent Assay (ELISA) method to determine the active constituents precisely and accurately in formulations to affirm broad spectrum safety and clinical efficacy.
Methods: Improved precision ELISA methodology was utilized to determine the amount of undenatured type II collagen extracted from chicken sternum cartilage. A commercially available Chondrex Collagen Detection Kit was used to determine the number of epitope (antigenic determinant) sites on the three-dimensional tightly-folded structured collagen. Time and temperature were set at ≥16 h or preferably within the range of 16 h to 24 h and at room temperature
Results: The results obtained from this improved ELISA method strongly supported the accuracy and validity, which correlates very well with the results of our earlier clinical studies, revealing the efficacy of undenatured type II collagen concentrations used. Furthermore, the modified ELISA method, designed by our team, revealed consistent and reproducible results on the basis of counting the epitope sites in undenatured type II collagen (NEXT-II®) of commercial batches
Conclusion: Using this precisely modified ELISA method gave 8% of undenatured type II collagen in NEXT-II®, resulting in 3.2 mg in 40.0 mg of NEXT-II®. It also confirmed that administration of 3.2 mg of undenatured type II collagen a day, both in open-label and randomized clinical trials, was safe and efficacious for joint pain, flexibility and mobility, and motor function.
Keywords: Undenatured type II collagen, NEXT-II®, ELISA method, Pepsin
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