INTRODUCTION:

Cyanocobalamin, an important form of vitamin B12, plays an essential role in human health and is, therefore, the basis for the formulation of various pharmaceuticals and nutritional supplements.¹ However, the susceptibility of cyanocobalamin to photodegradation poses significant challenges to its stability and effectiveness. This comprehensive review addresses this issue and explores the potential of cyclodextrins as a protectant against photodegradation. Examining the effectiveness of polymer cyclodextrin.

This review summarizes recent advances in this field and focuses on the mechanisms by which polymeric cyclodextrins protect cyanocobalamin from photodegradation. By investigating the structural properties and encapsulation ability of cyclodextrins. Furthermore, we evaluated the different methods and analytical techniques used to assess and quantify this stability, providing practical insights for pharmaceutical and nutraceutical applications². Photostability is the ability of a substance to resist degradation or breakdown when exposed to light. Cyanocobalamin, a vitamin B12, is particularly sensitive to degradation by light. When cyanocobalamin molecules are exposed to light, especially in the ultraviolet (UV) range, they can trigger photodegradation processes that can significantly reduce their potency and effectiveness. The main mechanism behind the photodegradation of cyanocobalamin involves photochemical reactions triggered by the absorption of light energy³. Ultraviolet light, in particular can break the chemical bonds within the cyanocobalamin molecule. These reactions often result in the formation of free radicals and the breakdown of the molecule into small fragments, ultimately leading to the loss of its biological activity.

Example for kinetics of cyanocobalamin degradation:

This review investigated the photolysis of cyanocobalamin (B12) and hydroxocobalamin (B12b) in the presence of cysteine (CYS). The reactions follow first-order kinetics, with first-order constants and rate constants at pH 2.0-12. B12b-CYS interaction has greater susceptibility to degradation, with k2 values nearly twice that of B12- CYS. The k2-pH profiles showed CYS ionization affected reaction and rates. Quantum yields range from 0.001 to 0.009 and 0.007 to 0.013, respectively.²

Photodegradation Effects of Cyanocobalamin:

· Loss of Potency: Degradation reduces active form of cyanocobalamin, affecting its effectiveness in physiological functions.

· Reduced Shelf Life: Exposure to light accelerates degradation, reducing product shelf life.

· Changes in Molecular Structure: Photolysis alters cyanocobalamin's structure, producing less bioavailable forms.

· Protective Measures: Polymer cyclodextrin encapsulates cyanocobalamin molecules, reducing exposure to light and minimizing photodegradation. Vitamin B12 is a stable metal complex with a cobalt ion in the +3-oxidation state, coordinated with nitrogen atoms and residue groups. It forms vitamers soluble in organic solvents, allowing for vitamin B12 catalysis. The catalytic activity of vitamin B12 is due to its ability to form weak cobalt-carbon bonds. Understanding photodegradation effects on cyanocobalamin is crucial for maintaining its stability and efficacy in pharmaceutical and nutraceutical applications^3,12.

Synergistic Interaction between Cyclodextrin and Cyclocobalamin:

· Cyclodextrin and cyanocobalamin form a protective shield, limiting exposure to light and reducing photo degradation.

· Cyclodextrin acts as a light-blocking barrier, preventing direct contact with cyanocobalamin molecules.

· Cyclodextrin provides a stable microenvironment within the cyclodextrin cavity, protecting cyanocobalamin from degradation factors like oxygen and other reactive species.

· The combined action of cyclodextrin and cyanocobalamin extends the shelf life of cyanocobalamin-containing formulations by reducing photodegradation.

· This interaction may show a protective alliance, ensuring long-term efficacy and ease of use in pharmaceutical and nutraceutical applications⁴.

· Polymeric cyclodextrins are compounds formed by joining multiple cyclodextrin molecules through covalent bonds or physical interactions. These cyclic oligosaccharides have a hydrophobic cavity and a hydrophilic exterior, allowing guest molecules like drugs or vitamins to be encapsulated within their cavities. When polymerized, these cyclodextrins exhibit enhanced encapsulation ability, producing larger, more complex structures with multiple cavities.⁵

Polymeric Cyclodextrins in Cyclocobalamin Encapsulation:

· Improved Encapsulation Efficiency: Larger structure and multiple cavities allow for higher encapsulation efficiency, enabling more molecules like cyanocobalamin to be encapsulated.

· Improved Stability: Expanded network provides a safer environment, increasing stability of encapsulated molecules.

· Customized Features: Chemical modifications can alter size, shape, and surface properties, allowing for customization for various applications.

· Controlled Release: Encapsulated compounds can be released in response to specific triggers like pH, temperature, or enzyme activity.

^·Effective carriers in cyanocobalamin encapsulation due to increased efficiency, stability, and potential for tailored formulations.⁶

Different pathways of degradation:

Cyanocobalamin is sensitive to light, so exposure to light can lead to decomposition processes, especially in the presence of ultraviolet light. Several mechanisms may be involved in the degradation pathway of cyanocobalamin.^7-10

Degradation Pathways of Cyanocobalamin:

· Cobalt-Carbon Bond Breaking: UV light can break the cobalt-carbon bond in cyanocobalamin, releasing cyanide ions and fragmenting molecules.¹¹

· Formation of Free Radicals: UV light can induce free radicals within cyanocobalamin molecules, leading to chain reactions and degradation products.

· Isomerization and Decomposition: UV light can trigger isomerization processes within cyanocobalamin, altering atom spatial arrangement and causing degradation.

· Oxidative Degradation: UV light can accelerate oxidative degradation, causing oxidative damage and degradation.

· Loss of Biological Activity: Understanding these degradation pathways can help implement protective measures and develop strategies to maintain cyanocobalamin stability.^7-10

ANALYTICAL TECHNIQUES:^13-22

Some commonly used analytical techniques mentioned below are used to assess the stability and degradation of compounds such as cyanocobalamin.

1. High-Performance Liquid Chromatography (HPLC): Analytical Techniques for Cyanocobalamin Stability and Degradation. high-Performance Liquid Chromatography (HPLC): Separates, identifies, and quantifies compounds in mixtures. Used to detect degradation products, monitor changes in cyanocobalamin concentrations, and assess stability over time. Chromatographic conditions: Mobile phases were 5mM ammonium formate in water acidified with 0.05% formic acid (A) and acetonitrile with 0.3% formic acid (B).

2. UV-visible spectroscopy: Measures light absorption in ultraviolet and visible regions of the electromagnetic spectrum. Spectrometric assay of B12–CYS and B12a–CYS/B12b–CYS solutions: B12–CYS photolyzed solutions showed an absorption maximum at 550nm, similar to B12 alone. B12a–CYS photolyzed solutions showed an absorption maximum at 531nm, possibly due to photochemical interaction with CYS.

3. Mass Spectrometry (MS): MS is used to identify and quantify compounds based on their mass-to-charge ratio. This is valuable for the characterization of the degradation products formed during the degradation of cyanocobalamin and provides insight into the degradation pathway.

4. Fourier Transform Infrared Spectroscopy (FTIR): FTIR analyses the infrared absorption or emission of molecules and helps identify functional groups and structural changes in cyanocobalamin caused by the degradation process.

5. Differential Scanning Calorimetry (DSC): DSC measures the heat flow associated with thermal transitions in materials. It is used to evaluate changes in the thermal properties of cyanocobalamin due to degradation or interaction with other compounds.

6. Accelerated stability studies: These studies expose cyanocobalamin-containing formulations to increased stress conditions, such as elevated temperatures and humidity, to simulate long-term storage over short periods. Changes in the properties of cyanocobalamin under these conditions provide information regarding its stability.

7. Gas Chromatography (GC): GC is used for the separation and analysis of volatile compounds. It may be useful for detecting volatile degradation products of cyanocobalamin.

8. Titration Techniques: Quantitative analysis can be performed using various titration methods to assess changes in cyanocobalamin concentration due to degradation.

These techniques allow researchers and analysts to assess the stability, degradation pathways, and time course of cyanocobalamin formulations, providing important data to optimize storage conditions and formulation strategies.

MATERIALS AND METHODS:^23-27

Materials: Vitamin, cyclodextrin, required excipents, respective equipments.

Methods:

These are the methods used to prepare vitamin B12-cyclodextrin complexes or formulations.^45- ⁴⁶

1. Co-precipitation or Co-evaporation:

Vitamin B12 and cyclodextrin can be dissolved together in a suitable solvent and then co-precipitated or co- evaporated to form the complex. The solvent is removed under controlled conditions to obtain the solid complex.

2. Kneading Method:

This involves mixing vitamin B12 and cyclodextrin in a mortar or a kneading machine with a minimal amount of solvent. The mixture is kneaded thoroughly until a homogeneous mass is obtained, and then the solvent is evaporated.

3. Spray Drying:

Vitamin B12 and cyclodextrin solutions are atomized and sprayed into a chamber under controlled temperature and pressure conditions. The solvent evaporates rapidly, leaving behind dry particles of the complex.

4. Freeze-Drying (Lyophilization):

A solution containing vitamin B12 and cyclodextrin is frozen and then subjected to vacuum conditions to remove the solvent through sublimation, leaving behind a dry complex.

5. Inclusion Complex Formation by Solvent Method:

Vitamin B12 and cyclodextrin are dissolved in a suitable solvent and left for a certain period to allow the complex formation. The solvent is then evaporated to obtain the complex.

6. Encapsulation Techniques:

Utilizing techniques like microencapsulation or nanoencapsulation, where vitamin B12 is encapsulated within cyclodextrin micro/nanoparticles to form a stable complex.

7. Co-grinding or Co-milling:

Vitamin B12 and cyclodextrin are mechanically ground together to form a complex. This method requires high shear forces and can be performed using ball milling or similar techniques.

Each method offers its advantages and may be selected based on factors such as scalability, efficiency, the physical state desired for the final product, and the properties of the substances being used. The choice of method often depends on the specific requirements of the intended application and the characteristics of the vitamin B12-cyclodextrin complex needed.

CONCLUSION:

In conclusion, the preservation of cyanocobalamin's stability against light-induced degradation stands as a critical endeavor in the pharmaceutical and nutraceutical realms. This comprehensive review has underscored the pronounced photosensitivity of cyanocobalamin, elucidating the multifaceted degradation pathways initiated upon exposure to light, notably UV radiation.Central to the quest for stability enhancement, polymer cyclodextrins emerge as pivotal guardians, exhibiting remarkable efficacy in shielding cyanocobalamin from photodegradation. Their encapsulation prowess forms a protective barrier, sequestering cyanocobalamin molecules and curbing their direct exposure to light. This synergistic interaction fosters a stabilized microenvironment, preserving the structural integrity and potency of cyanocobalamin. Moreover, this review accentuates the significance of analytical methodologies, encompassing HPLC, UV-visible spectroscopy, and mass spectrometry, among others, as indispensable tools in assessing cyanocobalamin's stability and unraveling degradation pathways.³⁹ As the understanding of cyanocobalamin's photosensitivity deepens, the utilization of polymer cyclodextrins stands as a promising avenue in formulating strategies to extend the shelf life, efficacy, and utility of cyanocobalamin-containing products. By harnessing the protective capabilities of polymer cyclodextrins and leveraging analytical insights,this review advocates for a concerted effort to fortify the stability of cyanocobalamin, ensuring its sustained therapeutic relevance across pharmaceutical and nutraceutical domains.In amalgamation, the alliance between polymer cyclodextrins and cyanocobalamin heralds a promising trajectory in fortifying stability against light-induced degradation, thus amplifying the potential for enhanced formulations in diverse applications. The complexation of vitamin B12 with cyclodextrins enhances its stability, solubility, and bioavailability. Techniques like co-precipitation, kneading, spray drying, freeze-drying, solvent- based complexation, encapsulation, and co-grinding create stable complexes. These methods can be used for oral supplements, injections, topical treatments, and more. The choice of formulation method depends on scalability, efficiency, and final product use. This conclusion encapsulates the significance of cyanocobalamin stability, the protective role of polymer cyclodextrins, the relevance of analytical techniques, and the potential impact on pharmaceutical and nutraceutical applications. Adjustments can be made to tailor the conclusion further to suit specific aspects or emphasis points within your review article.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

The authors would like to thank Sarada Vilas College of Pharmacy for their support and guidence.

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Received on 20.02.2024 Revised on 13.05.2024

Accepted on 20.07.2024 Published on 28.01.2025

Available online from February 27, 2025

Research J. Pharmacy and Technology. 2025;18(2):907-911.

DOI: 10.52711/0974-360X.2025.00133

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

P. K. Kulkarni, Venkatesh, K Hanumanthachar Joshi, A Noorain KM*