Functional Characterization of Undenatured Type II Collagen Supplements: Are They Interchangeable?

Figures & data

Table 1. Type II Collagen assay methods and product specifications.

	CII X	CII Y
	Native type II collagen from avian sternum (breastbone)	Undenatured type II collagen from chicken sternum (breastbone)
Total collagen – method	EP 2.2.56, Hydrolysis method 1, Derivatization method 7	Pepsin & elastase digestion; AAA analysis
Product specification^a	≥25%	≥25%
Measured amount^a	48%	31%
Undenatured collagen – method	Chymotrypsin digestion, EP 2.2.56, Hydrolysis method 1, Derivatization method 7	Pepsin & elastase digestion; ELISA assay
Product specification^a	≥4%	≥3%
Measured amount^a	18%	9%

^a Data from product Certificates of Analysis.

Table

Code	Name	Lot	Manufacturer
CII-A	Colflex capsules	No. L190520A	Mariol Industrial Ltda, Brazil
CII-B	Protena Soft Gel Capsules	No. L031R0119	ACHE, Brazil
CII-C	Motilex Capsules	No. L19080122	Apsen Pharma, Brazil
CII-D	NT2 Collagen Capsules	No. M18270C01	Quality Supplements & Vitamins Inc, USA
CII-E	NC2 Collagen Capsules	No. 18K05A	Trenker S.A. Laboratoires, Germany
CII- F	UC-II Capsules	No. 1808010	Lonza CHI, USA

Each of the brands was specified on the label to contain 40 mg collagen per capsule.

Figure 1. CII-Y has a high “L” reading compared to CII-X indicating it is slightly lighter, while CII-X is higher in “a” and “b” measurements indicating that it is more brownish.

Figure 2. Visual Appearance of CII-X and CII-Y powders. CII-Y is off-white and contains some coarse particles, while powder CII-X is light brown and displays coarser particles.

Table 2. Physical Characteristics of Collagen Powders.

Samples		CII-X	CII-Y
Visual appearance		Light brown, more course particles	Off-white, some coarse particles
Odor		slight odor	slightly less odor
Hunter Color	L (black is 0 to white is 100)	76.81	89.77
	a (green to red)	5.16	1.02
	b (blue to yellow)	18.94	14.25
Tapped density	(g/cc)	0.741	0.696
Particle size distribution	Measured size distribution <1 nm–6 μm	Coarse particles sedimented fast. Bimodal distribution with similar proportion of fines (∼1 μm) and similar tendency of overs to sediment as CII-Y, but larger variability in the 7–10 μm range.	Coarse particles sedimented fast. Bimodal distribution with similar proportion of fines (∼1 μm) and similar tendency of overs to sediment as CII-X, but less variability in 7–10 μm range.
Moisture (by LOD)	%	7.97	6.55
pH (5% solution)		7.488	6.094
Nitrogen	g/100 g	12.3	8.13
Ash	g/100 g	10.8	35

Figure 3. Solubility of collagen powders. CII-X was large and lumpy in appearance. CII-Y was finer, smaller, and homogeneous.

Table 3. Fatty acid profile.

Fatty Acid Profile	CII-X (g/100 g)	CII-Y (g/100 g)
Saturated fatty acids	0.044	0.671
Total cis Unsaturated fatty acids	0.026	1.060
Monounsaturated fatty acids	0.026	0.819
Polyunsaturated fatty acids	<0.007	0.240
Trans fatty acids	<0.007	0.035
Total fatty acids	0.073	1.850

Table 4. Quality profiles for collagen powders.

	CII-X	CII-Y
Heavy metals by ICP MS (ppb)
Arsenic	<10.0	<25.0
Cadmium	<5.00	<12.5
Chromium	567	413
Copper	1190	720
Lead	20.7	14.6
Mercury	6.72	<12.5
Zinc	22,400	10,400
Microbiological assays
Aerobic plate count	6600 CFU/g	50 CFU/g
E. coli	Absent/10 g	Absent/10 g
Enterobacteriaceae	>100 but <1000 MPN/g	<10 MPN/g
Salmonella	Absent/10 g	Absent/10 g
S. aureus	Absent/10 g	Absent/10 g
Yeast and mold	<10 CFU/g	<10 CFU/g

Figure 4. TEM micrographs showing difference in the appearance of CII-X and CII-Y powders (500 nM). CII-Y samples shows fibrils typical of type II collagen. CII-X failed to present with the typical fibril structures characteristic of intact type II collagen fibers.

Table 5. Type II collagen raw materials: total collagen and undenatured type II collagen.

Collagen raw material	Lot no.	Total collagen (%)	%Undenatured type II collagen
CII-X	19/0010	25.56	0.24
CII- X1	18/0008	33.36	0.12
CII- X2	20/0005	37.6	0.019
CII-Y	19FS211801	35.0	8.00
CII-Y1	19FS093321	21.9	9.90
CII-Y2	19FS122324	30.6	10.11

Table 6. ELISA analysis of a mixture of raw materials.

Collagen	Lot no.	Wt.% of solids	Average wt%
CII-Y1	19FS093321	6.52	6.52
CII- X1	18/0008	0.009
1:1 mixture		3.21	3.94
1:1 mixture		4.58

Table 7. Capsules: total collagen and undenatured type II collagen.

Collagen type II capsules	Lot no.	Undenatured type II collagen per capsule (mg; based on ELISA assay)
CII-A	456-F44F	0.409
CII-B	L031R0119	0.371
CII-C	L1908122	0.618
CII-D	M18270C1	0.841
CII-E	18K05A	0.242
CII-F	180897	2.352