Detection of 7-Dehydrocholesterol and Vitamin D3 Derivatives in Honey

Abstract

20(S)-Hydroxyvitamin D3 (20(OH)D3) is an endogenous metabolite produced by the action of CYP11A1 on the side chain of vitamin D3 (D3). 20(OH)D3 can be further hydroxylated by CYP11A1, CYP27A1, CYP24A1 and/or CYP27B1 to several hydroxyderivatives. CYP11A1 also hydroxylates D3 to 22-monohydroxyvitamin D3 (22(OH)D3), which is detectable in the epidermis. 20-Hydroxy-7-dehydrocholesterol (20(OH)-7DHC) has been detected in the human epidermis and can be phototransformed into 20(OH)D3 following the absorption of ultraviolet B (UVB) energy by the B-ring. 20(OH)D3 and its hydroxyderivatives have anti-inflammatory, pro-differentiation and anti-proliferative effects, comparable to 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Since cytochromes P450 with 20- or 25-hydroxylase activity are found in insects participating in ecdysone synthesis from 7-dehydrocholesterol (7DHC), we tested whether D3-hydroxyderivatives are present in honey, implying their production in bees. Honey was collected during summer in the Birmingham area of Alabama or purchased commercially and extracted and analyzed using LC-MS. We detected a clear peak of m/z = 423.324 [M + Na]+ for 20(OH)D3 corresponding to a concentration in honey of 256 ng/g. We also detected peaks of m/z = 383.331 [M + H − H2O]+ for 20(OH)-7DHC and 25(OH)D3 with retention times corresponding to the standards. We further detected species with m/z = 407.329 [M + Na]+ corresponding to the RT of 7DHC, D3 and lumisterol3 (L3). Similarly, peaks with m/z = 399.326 [M + H − H2O]+ were detected at the RT of 1,25(OH)2D3 and 1,20-dihydroxyvitamin D3 (1,20(OH)2D3). Species corresponding to 20-monohydroxylumisterol3 (20(OH)L3), 22-monohydroxyvitamin D3 (22(OH)D3), 20,23-dihydroxyvitamin D3 (20,23(OH)2D3), 20,24/25/26-dihydroxyvitamin D3 (20,24/25/26(OH)2D3) and 1,20,23/24/25/26-trihydroxyvitamin D3 (1,20,23/24/25/26(OH)3D3) were not detectable above the background. In conclusion, the presence of 7DHC and D3 and of species corresponding to 20(OH)-7DHC, 20(OH)D3, 1,20(OH)2D3, 25(OH)D3 and 1,25(OH)2D3 in honey implies their production in bees, although the precise biochemistry and photochemistry of these processes remain to be defined.

COVID‐19 and Vitamin D: A lesson from the skin

“…[ 12,13,14,15 ] In addition, 20(OH)D 3 has been detected in the honey, which defines it as a natural product. [ 16 ] CYP11A1 is expressed not only in adrenals, placenta and gonads but also in immune cells and other peripheral organs. [ 17 ]…”

“…[12,13,14,15] In addition, 20(OH)D 3 has been detected in the honey, which defines it as a natural product. [16] CYP11A1 is expressed not only in adrenals, placenta and gonads but also in immune cells and other peripheral organs. [17] Both 1,25(OH) 2 D 3 -and non-calcemic CYP11A1-derived metabolites use various, although partially overlapping, mechanisms in enacting their anti-inflammatory and antioxidative effects Figure 1B. 1,25(OH) 2 D 3 mediates many of its anti-inflammatory and anti-microbial effects through the vitamin D receptor (VDR).…”

20S-Hydroxyvitamin D3, a Secosteroid Produced in Humans, Is Anti-Inflammatory and Inhibits Murine Autoimmune Arthritis

“…This suggests that 20 S (OH)D3 may also be produced by the immune cells. Furthermore, 20 S (OH)D3 has been detected in honey identifying it as a natural product ( 42 ). The mechanisms by which 20 S (OH)D3 downregulates arthritis severity in the CIA model is likely related to reduction in CD4 + T cells, CD19 + B cells, anti-CII antibodies, and maintenance of CD4 + CD24 + FoxP3 + Tregs.…”

“…The main and first product of the pathway, 20S(OH)D3, in contrast to 25(OH)D3 or 1,25(OH) 2 D3, is nontoxic (i.e., noncalcemic and nontoxic to the hematopoietic system, liver, kidney, and heart) at doses as high as 60 µg/kg, while 25(OH)D3 or 1,25(OH) 2 D3 induce hypercalcemia at doses ≤2 µg/kg (38)(39)(40)(41). Finally, 20S(OH)D3 is produced in vivo, being present in human serum at 1/20 th the concentration of classical 25(OH)D3 (31) and has also been detected in honey (42). The cytochrome SCC enzymes are unusual in that they hydroxylate vitamin D3 to produce 20S-hydroxyvitamin D3 [20(OH)D3] rather than cleaving the side chains of D3 (43).…”

The Impact of Vitamin D on Skin Aging

“…The major CYP11A1-derived vitamin D 3 products are 20(OH)D 3 and 20,23(OH) 2 D 3 [23,139,147,148]. In addition, 20(OH)D 3 can be defined also as a natural product because its presence in the honey [149]. The L 3 can also be metabolized to biologically active derivatives [150][151][152], which are not recognized by the 7-DHC reductase [153].…”