Development and validation of a spectrofluorimetric method for the analysis of tolfenamic acid in pure and tablet dosage form.

Tolfenamic acid (TA) is commonly used in humans and animals because of its anti-inflammatory, antipyretic, and analgesic effects. So far, no study has been carried out to develop a validated spectrofluorimetric method for determination of TA. Therefore, the present study aimed to develop and validate a simple, accurate, rapid, economical, and precise spectrofluorimetric method to assay TA in its pure and dosage forms, and also in degraded solutions. The fluorimetric method had higher sensitivity compared with the spectrophotometric and high-performance liquid chromatography methods and could determine the drug at the microgram level. Optimum pH of TA for maximum fluorescence intensity was 3, and its two pKa values were calculated as 1.95 and 4.05. The proposed method was validated according to the guidelines of the International Council for Harmonisation, and parameters such as linearity, range, accuracy, precision, sensitivity, robustness, specificity, and solution stability were tested. Stress-induced degradation studies on TA did not affect the accuracy and precision of the proposed method. The results obtained indicated that the method was linear over the concentration range 0.2-0.9 × 10-3 M with good accuracy, precision, and robustness for assay of TA in its pure and its tablet dosage forms and was comparable statistically with the British Pharmacopoeia method.

Stability-indicating spectrofluorimetric method for the assay of riboflavin and photoproducts: Kinetic applications.

A stability-indicating spectrofluorimetric method has been developed for the simultaneous assay of riboflavin (RF) and photoproducts, formylmethylflavin (FMF), lumichrome (LC) and lumiflavin (LF) in aqueous solution. The method is based on the extraction of LC formed in acid solution and LC and LF formed in alkaline solution with chloroform at pH 2.0 and their assay by fluorescence measurements at 478 and 530 nm, respectively. The aqueous phase, on readjustment of the pH to 6.5, is used to extract FMF with chloroform and its assay is carried out at 530 nm. The aqueous phase is then used for the assay of RF at 530 nm. The proposed method gives more accurate results for the assay of RF compared to those of the United States Pharmacopeia (USP) spectrofluorimetric method which does not take into account the presence of RF photoproducts having similar fluorescence characteristics. The proposed method along with the USP method has been applied to the study of the kinetics of photolysis of RF, assay of stored commercial vitamin preparations and their radiated samples. The results show that the USP method does not distinguish between the fluorescence of RF and its photoproducts, and, therefore, gives erroneous results with about 11% excess in the quantity of the vitamin compared to that of the proposed method. This is due to the interference of the fluorescence of photoproducts in the assay of RF. The method has been validated for various analytical parameters according to the guideline of the International Council for Harmonization (ICH).

Multicomponent spectrofluorimetric method for the assay of carboxymethylflavin and its hydrolytic products: kinetic applications.

The simultaneous assay of carboxymethylflavin (CMF), an intermediate in the photolysis of riboflavin, and its hydrolytic side-chain cleavage products, lumichrome (LC) (acid solution) and LC and lumiflavin (LF) as well as isoalloxazine ring cleavage products, 1,2-dihydro-1-methyl-2-keto-3-quinoxaline carboxylic acid (KA) and 1,2,3,4-tetrahydro-1-methyl-2,3-dioxo-quinoxaline (DQ) (alkaline solution) has been carried out by a multicomponent spectrofluorimetric method. The method is based on the adjustment of pH of the degraded solutions to 2.0 and extraction of LC and LF with chloroform. The chloroform extract is evaporated to dryness under reduced pressure, the residue dissolved in pH 6.5 citro-phosphate buffer and LC and LF determined at their fluorescence maxima at 478 and 530 nm, respectively. The pH of the aqueous phase is re-adjusted to 6.5 and the solution used for the determination of CMF, KA and DQ at the wavelengths of 530, 443 and 420 nm, respectively. The proposed method has been validated according to ICH guidelines. The calibration curves for CMF and its hydrolytic products are linear in the concentration range of 0.5-5.0 × 10-6  M. The mean recovery ranges from 99.0-102.0% with relative standard deviation (RSD) of 0.19-0.99%. The limit of detection (LOD) and the limit of quantification (LOQ) are in the range of 1.17-1.78 × 10-7  M and 3.55-5.40 × 10-7  M, respectively. The uniformity of molar balance of CMF and degradation products during hydrolytic reactions indicates the accuracy of the proposed method for the spectrofluorimetric assay of the compounds. It has been applied to study the kinetics of hydrolytic reactions of CMF.

Effect of pH, polymer concentration and molecular weight on the physical state properties of tolfenamic acid.

Tolfenamic acid (TA) has been transformed from crystalline to amorphous state through freeze-drying by using varying ratios of polyacrylic acid (PA) at various pH values. The characterization of the films has been carried out using X-ray diffraction, differential scanning calorimetry, Fourier transform infrared (FTIR) spectrometry and scanning electron microscopy. The results showed a gradual change in the solid state properties of TA and a complete transformation into its amorphous form in 1:8, 1:4, 1:2 and 1:1 ratios at pH 3, 4, 5 and 6, respectively. FTIR spectrometry reveals the formation of a yellow polymorphic form of TA. Polymer molecular weight has also been observed to affect the drug transformation and interaction as the low molecular weight PA (Mw ∼ 1800) was found to be most effective followed by its medium (Mv ∼ 450 000) and high molecular weight (Mv ∼ 3 000 000) forms. No signs of recrystallization in the TA-PA films were noted during the 12-week storage period. PA of low molecular weight has also been found more effective in inhibiting the recrystallization of the melt upon cooling thus proving a valuable polymer in producing stable amorphous solid dispersions of TA.

Solvent Effect on the Photolysis of Riboflavin.

The kinetics of photolysis of riboflavin (RF) in water (pH 7.0) and in organic solvents (acetonitrile, methanol, ethanol, 1-propanol, 1-butanol, ethyl acetate) has been studied using a multicomponent spectrometric method for the assay of RF and its major photoproducts, formylmethylflavin and lumichrome. The apparent first-order rate constants (k obs) for the reaction range from 3.19 (ethyl acetate) to 4.61 × 10(-3) min(-1) (water). The values of k obs have been found to be a linear function of solvent dielectric constant implying the participation of a dipolar intermediate along the reaction pathway. The degradation of this intermediate is promoted by the polarity of the medium. This indicates a greater stabilization of the excited-triplet states of RF with an increase in solvent polarity to facilitate its reduction. The rate constants for the reaction show a linear relation with the solvent acceptor number indicating the degree of solute-solvent interaction in different solvents. It would depend on the electron-donating capacity of RF molecule in organic solvents. The values of k obs are inversely proportional to the viscosity of the medium as a result of diffusion-controlled processes.

Effect of ascorbic acid on the degradation of cyanocobalamin and hydroxocobalamin in aqueous solution: a kinetic study.

The degradation kinetics of 5 × 10(-5) M cyanocobalamin (B12) and hydroxocobalamin (B12b) in the presence of ascorbic acid (AH2) was studied in the pH range of 1.0-8.0. B12 is degraded to B12b which undergoes oxidation to corrin ring cleavage products. B12b alone is directly oxidized to the ring cleavage products. B12 and B12b in degraded solutions were simultaneously assayed by a two-component spectrometric method at 525 and 550 nm without interference from AH2. Both degrade by first-order kinetics and the values of the rate constants at pH 1.0-8.0 range from 0.08 to 1.05 × 10(-5) s(-1) and 0.22-7.62 × 10(-5) s(-1), respectively, in the presence of 0.25 × 10(-3) M AH2. The t 1/2 values of B12 and B12b range from 13.7 to 137.5 h and 2.5-87.5 h, respectively. The second-order rate constants for the interaction of AH2 with B12 and B12b are 0.05-0.28 × 10(-2) and 1.10-30.08 × 10(-2) M(-1) s(-1), respectively, indicating a greater effect of AH2 on B12b compared to that of B12. The k obs-pH profiles for both B12 and B12b show the highest rates of degradation around pH 5. The degradation of B12 and B12b by AH2 is affected by the catalytic effect of phosphate ions on the oxidation of AH2 in the pH range 6.0-8.0.

Photodegradation of moxifloxacin in aqueous and organic solvents: a kinetic study.

The kinetics of photodegradation of moxifloxacin (MF) in aqueous solution (pH 2.0-12.0), and organic solvents has been studied. MF photodegradation is a specific acid-base catalyzed reaction and follows first-order kinetics. The apparent first-order rate constants (kobs) for the photodegradation of MF range from 0.69 × 10(-4) (pH 7.5) to 19.50 × 10(-4) min(-1) (pH 12.0), and in organic solvents from 1.24 × 10(-4) (1-butanol) to 2.04 × 10(-4) min(-1) (acetonitrile). The second-order rate constant (k2) for the [H(+)]-catalyzed and [OH(-)]-catalyzed reactions are 6.61 × 10(-2) and 19.20 × 10(-2) M(-1) min(-1), respectively. This indicates that the specific base-catalyzed reaction is about three-fold faster than that of the specific acid-catalyzed reaction probably as a result of the rapid cleavage of diazabicyclononane side chain in the molecule. The kobs-pH profile for the degradation reactions is a V-shaped curve indicating specific acid-base catalysis. The minimum rate of photodegradation at pH 7-8 is due to the presence of zwitterionic species. There is a linear relation between kobs and the dielectric constant and an inverse relation between kobs and the viscosity of the solvent. Some photodegraded products of MF have been identified and pathways proposed for their formation in acid and alkaline solutions.

Photo, thermal and chemical degradation of riboflavin.

Riboflavin (RF), also known as vitamin B2, belongs to the class of water-soluble vitamins and is widely present in a variety of food products. It is sensitive to light and high temperature, and therefore, needs a consideration of these factors for its stability in food products and pharmaceutical preparations. A number of other factors have also been identified that affect the stability of RF. These factors include radiation source, its intensity and wavelength, pH, presence of oxygen, buffer concentration and ionic strength, solvent polarity and viscosity, and use of stabilizers and complexing agents. A detailed review of the literature in this field has been made and all those factors that affect the photo, thermal and chemical degradation of RF have been discussed. RF undergoes degradation through several mechanisms and an understanding of the mode of photo- and thermal degradation of RF may help in the stabilization of the vitamin. A general scheme for the photodegradation of RF is presented.

Photolysis of tolfenamic acid in aqueous and organic solvents: a kinetic study.

Tolfenamic acid (TA) is a non-steroidal anti-inflammatory drug that was studied for its photodegradation in aqueous (pH 2.0-12.0) and organic solvents (acetonitrile, methanol, ethanol, 1-propanol, 1-butanol). TA follows first-order kinetics for its photodegradation, and the apparent first-order rate constants (k obs) are in the range of 0.65 (pH 12.0) to 6.94 × 10-2 (pH 3.0) min-1 in aqueous solution and 3.28 (1-butanol) to 7.69 × 10-4 (acetonitrile) min-1 in organic solvents. The rate-pH profile for TA photodegradation is an inverted V (∧) or V-top shape, indicating that the cationic form is more susceptible to acid hydrolysis than the anionic form of TA, which is less susceptible to alkaline hydrolysis. The fluorescence behavior of TA also exhibits a V-top-shaped curve, indicating maximum fluorescence intensity at pH 3.0. TA is highly stable at a pH range of 5.0-7.0, making it suitable for formulation development. In organic solvents, the photodegradation rate of TA increases with the solvent's dielectric constant and solvent acceptor number, indicating solute-solvent interactions. The values of k obs decreased with increased viscosity of the solvents due to diffusion-controlled processes. The correlation between k obs versus ionization potential and solvent density has also been established. A total of 17 photoproducts have been identified through LC-MS, of which nine have been reported for the first time. It has been confirmed through electron spin resonance (ESR) spectrometry that the excited singlet state of TA is converted into an excited triplet state through intersystem crossing, which results in an increased rate of photodegradation in acetonitrile.

Studies on tolfenamic acid-chitosan intermolecular interactions: effect of pH, polymer concentration and molecular weight.

Solid-state properties of tolfenamic acid (TA) and its complexes with chitosan (CT) have been studied. Effect of medium pH, molecular weight of polymer and its different concentrations on these TA-CT complexes were studied in detail. Low and medium molecular weight CT have been used in different ratios at pH ranging from 4 to 6 and freeze-drying technique has been employed to modify the appearance of crystalline TA. Physical properties of the formed complexes have been studied by employing X-ray diffraction, differential scanning calorimetry and scanning electron microscopy; chemical structure has been studied using Fourier transform infrared spectroscopy. The results showed that both forms of the polymer exhibited complete conversion in 1:8 ratio at pH 4, 1:4 at pH 5 and 1:1 at pH 6 indicating a marked effect of pH on drug-polymer complexation. The percent crystallinity calculations indicated low molecular weight CT slightly more effective than the other form. No changes in the complexes have been observed during the 12 week storage under controlled conditions. Both forms of CT at different pH values indicated retardation of recrystallization in TA during cooling of the melt from 1:1 ratios exhibiting formation of strong intermolecular hydrogen bonding between the drug and the polymer.

Photodegradation and stabilization of betamethasone-17 valerate in aqueous/organic solvents and topical formulations.

The effects of solvent [acetonitrile, methanol, and acetonitrile/water mixture (20:80, v/v)], buffer concentration (phosphate buffer, pH 7.5), ionic strength and commonly employed adjuvants on the photodegradation of betamethasone-17 valerate in cream and gel formulations have been studied on exposure to UV light (300-400 nm). A validated high-performance liquid chromatography method has been used to determine the parent compound and its photodegraded products. The photodegradation data in the studied solvents showed greater decomposition of the drug in solvents with a lower dielectric constant. A comparatively higher rate of photodegradation was observed in the cream formulation compared to that for the gel formulation. The kinetic treatment of the photodegradation data revealed that the degradation of the drug follows first-order kinetics and the apparent first-order rate constants for the photodegradation reactions, in the media studied, range from 1.62 to 11.30×10(-3) min(-1). The values of the rate constants decrease with increasing phosphate concentration and ionic strength which could be due to the deactivation of the excited state and radical quenching. The second-order rate constant (k') for the phosphate ion-inhibited reactions at pH 7.5 has been found to be 5.22×10(-2) M(-1) s(-1). An effective photostabilization of the drug has been achieved in cream and gel formulations with titanium dioxide (33.5-42.5%), vanillin (21.6-28.7%), and butyl hydroxytoluene (18.2-21.6%).

Correction for irrelevant absorption in multicomponent spectrophotometric assay of riboflavin, formylmethylflavin, and degradation products: kinetic applications.

In the spectrophotometric assay of multicomponent systems involved in drug degradation studies, some minor or unknown degradation products may be present. These products may interfere in the assay and thus invalidate the results due to their absorption in the range of analytical wavelengths. This interference may be eliminated by the application of an appropriate correction procedure to obtain reliable data for kinetic treatment. The present study is based on the application of linear and non-linear irrelevant absorption corrections in the multicomponent spectrophotometric assay of riboflavin and formylmethylflavin during the photolysis and hydrolysis studies. The correction procedures take into account the interference caused by minor or unknown products and have shown considerable improvement in the assay data in terms of the molar balance. The treatment of the corrected data has led to more accurate kinetic results in degradation studies.

Pharmaceutical based cosmetic serums.

The growth and demand for cosmeceuticals (cosmetic products that have medicinal or drug-like benefits) have been enhanced for the last few decades. Lately, the newly invented dosage form, i.e., the pharmaceutical-based cosmetic serum has been developed and widely employed in various non-invasive cosmetic procedures. Many pharmaceutical-based cosmetic serums contain natural active components that claim to have a medical or drug-like effect on the skin, hair, and nails, including anti-aging, anti-wrinkle, anti-acne, hydrating, moisturizing, repairing, brightening and lightening skin, anti-hair fall, anti-fungal, and nail growth effect, etc. In comparison with other pharmaceutical-related cosmetic products (creams, gels, foams, and lotions, etc.), pharmaceutical-based cosmetic serums produce more rapid and incredible effects on the skin. This chapter provides detailed knowledge about the different marketed pharmaceutical-based cosmetic serums and their several types such as facial serums, hair serums, nail serums, under the eye serum, lip serum, hand, and foot serum, respectively. Moreover, some valuable procedures have also been discussed which provide prolong effects with desired results in the minimum duration of time after the few sessions of the serum treatment.

Nanosponges-based Drug Delivery System for the Cosmeceutical Applications of Stabilized Ascorbic Acid.

BACKGROUND: L-Ascorbic acid (AA) is a highly unstable compound, thus, limiting its use in pharmaceutical and cosmetic products, particularly at higher concentrations. OBJECTIVE: This study aimed to stabilize the highly sensitive molecule (AA) by encapsulating it in ß- cyclodextrin nanosponges (ß-CD NS) that can be used further in preparing cosmeceuticals products with higher AA concentrations and enhanced stability. METHODS: The NS has been synthesized by the melting method. The AA was encapsulated in ß-CD NS by the freeze-drying process. The prepared NS were characterized by FTIR spectrometry, SEM, Atomic Force Microscopy (AFM), zeta sizer, Differential Scanning Calorimetry (DSC), and the physical flow characteristics were also studied. The in vitro drug release was carried out on the Franz apparatus using a combination of two methods: sample & separate and dialysis membrane. The assay was performed using a validated spectrometric method. RESULTS: The entrapment efficiency of AA in ß-CD NS indicated a good loading capacity (83.57±6.35%). The FTIR, SEM, AFM, and DSC results confirmed the encapsulation of AA in ß-CD NS. The particle size, polydispersity index, and zeta potential results ascertained the formation of stabilized monodisperse nanoparticles. The physical flow characteristics showed good flow properties. Around 84% AA has been released from the NS in 4 h following the Korsmeyer-Peppas model. The AA-loaded NS remained stable for nine months when stored at 30±2°C/65±5% RH. CONCLUSION: It is concluded that the prepared NS can protect the highly sensitive AA from degradation and provide an extended-release of the vitamin. The prepared AA-loaded ß-CD NS can be used to formulate other cosmeceutical dosage forms with better stability and effect.

Analysis of Tolfenamic Acid using a Simple, Rapid, and Stabilityindicating Validated HPLC Method.

BACKGROUND: Tolfenamic acid (TA) belongs to the fenamates class of non-steroidal anti-inflammatory drugs. Insufficient information is available regarding the availa-bility of a reliable and validated stability-indicating method for the assay of TA. OBJECTIVE: A relatively simple, rapid, accurate, precise, economical, robust, and stability-indicating RP-HPLC method has been developed to determine TA in pure and tablet dosage forms. METHODS: The method was validated according to the ICH guideline, and parameters like linearity, range, selectivity, accuracy, precision, robustness, specificity, and solution stability were determined. TLC and FTIR spectrometry were used to ascertain the purity of TA. The specificity was determined with known impurities and after performing forced degradation, while the robustness was established by Plackett-Burman's experimental design. The mobile phase used for the analysis was acetonitrile and water (90:10, v/v) at pH 2.5. The detection of the active drug was made at 280 nm using a C18 column (tR = 4.3 min.). The method's ap-plicability was also checked for the yellow polymorphic form of TA. RESULTS: The results indicated that the method is highly accurate (99.39-100.80%), precise (<1.5% RSD), robust (<2% RSD), and statistically comparable to the British Pharmacopoeia method with better sensitivity and specificity. CONCLUSION: It was observed that the stress degradation studies do not affect the method's accuracy and specificity. Hence the proposed method can be used to assay TA and its tablet dosage form.

Photostability and interaction of ascorbic acid in cream formulations.

The kinetics of photolysis of ascorbic acid in cream formulations on UV irradiation has been studied using a specific spectrophotometric method with a reproducibility of ± 5%. The apparent first-order rate constants (k(obs)) for the photolysis of ascorbic acid in creams have been determined. The photoproducts formed in the cream formulations include dehydroascorbic acid and 2,3-diketogulonic acid. The photolysis of ascorbic acid appears to be affected by the concentration of active ingredient, pH, and viscosity of the medium and formulation characteristics. The study indicates that the ionized state and redox potentials of ascorbic acid are important factors in the photostability of the vitamin in cream formulations. The viscosity of the humectant present in the creams appears to influence the photostability of ascorbic acid. The results show that the physical stability of the creams is an important factor in the stabilization of the vitamin. In the cream formulations stored in the dark, ascorbic acid undergoes aerobic oxidation and the degradation is affected by similar factors as indicated in the photolysis reactions. The rate of oxidative degradation in the dark is about seventy times slower than that observed in the presence of light.

Photolysis of thiochrome in aqueous solution: A kinetic study.

The photolysis of thiochrome (THC), an oxidation product of thiamine (vitamin B1) (THE), used for its fluorimetric assay, has been studied in the pH range 7.0-12.0. THC undergoes photooxidation to oxodihydrothiochrome (ODTHC) which is oxidized to a non-fluorescent compound (OP1) on UV irradiation. The kinetics of the consecutive first-order reactions: THC→k1ODTHC→k2OP1, has been evaluated and the values of first-order rate constants, k1 (0.58-4.20 × 10-5, s-1) and k2 (0.05-2.03 × 10-5, s-1), at pH 7.0-12.0 have been determined. The rates of degradation of THC and ODTHC are enhanced with pH and the second-order rate constants k1' and k2' for the OH- ion-catalyzed reaction are in the range of 0.002-58.3 M-1 s-1. The quantum yields of the photolysis of THC and ODTHC in the pH range 7.0-12.0 have been determined. THC, ODTHC and OP1 have been identified by chromatographic, spectrometric and fluorimetric methods. THC and ODTHC have similar fluorescence characteristics and emit at 450 and 445 nm, respectively. THC, ODTHC and OP1 with distinct absorption maxima (370, 344 and 290 nm, respectively) have been determined by a newly developed and validated multicomponent spectrometric method during the photolysis reactions. The on-line formation of THC by the photooxidation of THE may lead to the degradation of THC and give erroneous results in the fluorimetric assay of THE. A scheme for the photolysis reactions of THC in aqueous solution is presented.

Effect of caffeine complexation on the photolysis of riboflavin in aqueous solution: a kinetic study.

The effect of caffeine complexation with riboflavin on the kinetics of riboflavin photolysis in the pH range 2.0-10.5 has been studied. The photolysis of riboflavin solutions (5x10(-5) M) was carried out in the presence of caffeine (0.5-2.5x10(-4) M) using a visible radiation source. A specific multicomponent spectrophotometric method was used for the determination of riboflavin and photoproducts in photolysed solutions. The apparent first-order rate constants (k) for the photolysis reactions range from 2.71x10(-4) to 4.26x10(-2) min(-1). The values of the rate constants decrease with increasing concentrations of caffeine indicating its inhibitory effect on the reactions. The second-order rate constants (k') for the caffeine inhibited reactions lie in the range of 0.13 to 5.10x10(-3) M(-1) min(-1). The log k-pH profiles for the photolysis reactions at various caffeine concentrations involve multiple steps indicating a gradual increase in the rate up to pH 10. The lower rates at pH 2.0 and 10.5 are due to the ionization of riboflavin as evident from fluorescence measurements. The k'-pH profile for the interaction of riboflavin with caffeine represents a bell-shaped curve in the pH range 3-6 followed by a sigmoid curve in the pH range 7-10. The inhibition of photolysis of riboflavin in the presence of caffeine is a result of the monomeric interaction and complex formation of caffeine with riboflavin. The photochemical interaction of riboflavin with caffeine suggests that a pH around 6 is most appropriate for the stabilization of the vitamin. At this pH the complex shows the highest stability constant.

Multicomponent spectrometric analysis of drugs and their preparations.

Pharmaceutical preparations may contain a single ingredient or multi-ingredients as well as excipients. In multicomponent systems, specific analytical methods are required to determine the concentrations of individual components in the presence of interfering substances. Ultraviolet and visible spectrometric methods have widely been developed for the analysis of drugs in mixtures and pharmaceutical preparations. These methods are based on ultraviolet and visible multicomponent analysis and chemometrics (multivariate data analysis). The commonly used chemometric methods include principal component analysis (PCA); regression involving classical least squares (CLS), partial least squares (PLS), inverse least squares (ILS), principal component regression (PCR), multiple linear regression (MLR), artificial neural networks (ANNs); soft independent modeling of class anthology (SIMCA), PLS-discriminant analysis (DA); and functional data analysis (FDA). In this chapter, the applications of multicomponent ultraviolet and visible, derivative, infrared and mass spectrometric and spectrofluorimetric methods to the analysis of multi-ingredient pharmaceutical preparations, biological samples and the kinetics of drug degradation have been reviewed. Chemometric methods provide an efficient solution to calibration problems in the analysis of spectral data for the simultaneous determination of drugs in multicomponent systems. These methods facilitate the assessment of product quality and enhance the efficiency of quality control systems.

Photolysis of carboxymethylflavin in aqueous and organic solvent: a kinetic study.

This is the first study on the photolysis of carboxymethylflavin (CMF), an intermediate in the photolysis of riboflavin (RF). CMF is photodegraded by removal of side-chain to lumichrome (LC) in acid solution and to LC and lumiflavin (LF) in alkaline solution. It also undergoes alkaline hydrolysis to 1,2-dihydro-1-methyl-2-keto-3-quinoxaline carboxylic acid (KA) and 1,2,3,4-tetrahydro-1-methyl-2,3-dioxoquinoxaline (DQ) by cleavage of isoalloxazine ring. CMF degrades to LC in organic solvents. The formation of LC in acid solution and organic solvents takes place by second-order reaction and those of LC, LF, KA and DQ in alkaline solution by first-order reactions. The values of second-order rate constants for the photolysis of CMF at pH 2.0 to 7.0 are in the range of 1.13 to 2.45 M-1 s-1 and those of first-order rate constants (k obs) at pH 8.0-12.0 from 1.53 to 4.18 × 10-4 s-1 and for the formation of photoproducts from 0.37 to 16.6 × 10-5 s-1. The photolysis of CMF is enhanced, with pH, in the alkaline region since the excited state is sensitive to alkaline hydrolysis. The photolysis and fluorescence quantum yields of CMF in aqueous and organic solvents have been reported. CMF and photoproducts have been assayed spectrofluorimetrically. The mode of CMF photolysis is discussed.