Form Of Pharmaceutical Ingredient Research Articles

Use of Essential Oils to Counteract the Phenomena of Antimicrobial Resistance in Livestock Species.

Antimicrobial resistance is an increasingly widespread phenomenon that is of particular concern because of the possible consequences in the years to come. The dynamics leading to the resistance of microbial strains are diverse, but certainly include the incorrect use of veterinary drugs both in terms of dosage and timing of administration. Moreover, the drug is often administered in the absence of a diagnosis. Many active ingredients in pharmaceutical formulations are, therefore, losing their efficacy. In this situation, it is imperative to seek alternative treatment solutions. Essential oils are mixtures of compounds with different pharmacological properties. They have been shown to possess the antibacterial, anti-parasitic, antiviral, and regulatory properties of numerous metabolic processes. The abundance of molecules they contain makes it difficult for treated microbial species to develop pharmacological resistance. Given their natural origin, they are environmentally friendly and show little or no toxicity to higher animals. There are several published studies on the use of essential oils as antimicrobials, but the present literature has not been adequately summarized in a manuscript. This review aims to shed light on the results achieved by the scientific community regarding the use of essential oils to treat the main agents of bacterial infection of veterinary interest in livestock. The Google Scholar, PubMed, SciELO, and SCOPUS databases were used for the search and selection of studies. The manuscript aims to lay the foundations for a new strategy of veterinary drug use that is more environmentally friendly and less prone to the emergence of drug resistance phenomena.

Cholinium-Based Ionic Liquids as Promising Antimicrobial Agents in Pharmaceutical Applications: Surface Activity, Antibacterial Activity and Ecotoxicological Profile.

Cholinium-based ionic liquids are compounds increasingly studied in pharmaceutics and biomedicine to enhance bioavailability in drug delivery systems and as bioactive ingredients in pharmaceutical formulations. However, their potential as antimicrobial agents has scarcely been investigated. Herein, we explored the antimicrobial activity of a series of surface-active cholinium-based ionic liquids (Chol-ILs). For this purpose, Chol-ILs with alkyl chains of 10-16 carbon atoms were synthesized and their self-assembly in aqueous medium was investigated. Subsequently, their antimicrobial activity against a panel of clinically relevant bacteria and their ability to eradicate MRSA and P. aeruginosa PAO1 biofilms was evaluated. Finally, we analyzed the ecotoxicological profile of Chol-ILs in terms of susceptibility to aerobic biodegradation and acute aquatic toxicity against D. magna and V. fisheri. Our results reveal that cholinium-based ILs with alkyl chain lengths ≥12 C show a broad spectrum of antibacterial activity. Their antimicrobial efficacy depends on their hydrophobicity, with the C14-C16 homologs being the most effective compounds. These ILs exhibit antimicrobial activity similar to that of imidazolium ILs and quaternary ammonium antiseptics. Moreover, the longer alkyl chain Chol-ILs are able to eradicate established biofilms at concentrations as low as 16-32 µg/mL. The biodegradation rate of cholinium-based ILs decreases with alkyl chain elongation. Our results reinforce the suitability of Chol-ILs as promising multifunctional compounds for application in pharmaceutical and biomedical formulation.

Comparative Genome-Wide Analysis of Two Caryopteris x Clandonensis Cultivars: Insights on the Biosynthesis of Volatile Terpenoids.

Caryopteris x Clandonensis, also known as bluebeard, is an ornamental plant containing a large variety of terpenes and terpene-like compounds. Four different cultivars were subjected to a principal component analysis to elucidate variations in terpenoid-biosynthesis and consequently, two representative cultivars were sequenced on a genomic level. Functional annotation of genes as well as comparative genome analysis on long read datasets enabled the identification of cultivar-specific terpene synthase and cytochrome p450 enzyme sequences. This enables new insights, especially since terpenoids in research and industry are gaining increasing interest due to their importance in areas such as food preservation, fragrances, or as active ingredients in pharmaceutical formulations. According to BUSCO assessments, the presented genomes have an average size of 355 Mb and about 96.8% completeness. An average of 52,090 genes could be annotated as putative proteins, whereas about 42 were associated with terpene synthases and about 1340 with cytochrome p450 enzymes.

Strategies for the Separation and Quantification of Non-Steroidal Anti- Inflammatory Drugs Using ZIC-HILIC-HPLC with UV Detection

Background: Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used worldwide for their analgesic, anti-inflammatory, and antipyretic properties. Objective: The article highlights the development of zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) methods to quantify three NSAIDs as active ingredients in pharmaceutical formulations simultaneously. Method: For NSAIDs analysis, two homemade ZIC-HILIC stationary phases were carried out by adding sulfobetaine monomers to polystyrene-divinylbenzene particles. The methods were developed by examining the effect of HILIC mode parameters like the kind of organic modifier (methanol or acetonitrile), acetonitrile content, pH and concentration of the acetate buffer in the eluent. Results: The experimental data exhibit the predominant mechanism of NSAIDs with two ZICHILIC stationary phases. The different chain lengths in the two ZIC-HILIC stationary phases are employed to investigate NSAIDs retention behaviour. The strategies were evaluated for their specificity, linearity, accuracy, and precision. The linear ranges were 0.01-5.0, 0.05-17.5 and 0.12-13.5 μg ml-1, LOD 0.005. 0.015 and 0.046 μg ml-1 and LOQ 0.015, 0.045 and 0.139 μg ml-1 for nimesulide, tenoxicam and diclofenac, respectively. Conclusion: They represent interactions with three NSAIDs, ZIC-HILIC columns, and mobile phase in a mixed-mode of hydrophilic and hydrophobic interactions. In contrast to the ZICHILIC- 1 stationary phase with a shorter chain length, the ZIC-HILIC-4 stationary phase with a longer chain length exhibited a longer retention time, lower detection and limit of quantitation for three NSAIDs.The verification findings demonstrate the suitability of the proposed ZIC-HILIC methods for quantifying NSAIDs in pharmaceutical formulations.

Formulation Excipients and Their Role in Insulin Stability and Association State in Formulation.

While excipients are often overlooked as the "inactive" ingredients in pharmaceutical formulations, they often play a critical role in protein stability and absorption kinetics. Recent work has identified an ultrafast absorbing insulin formulation that is the result of excipient modifications. Specifically, the insulin monomer can be isolated by replacing zinc and the phenolic preservative metacresol with phenoxyethanol as an antimicrobial agent and an amphiphilic acrylamide copolymer excipient for stability. A greater understanding is needed of the interplay between excipients, insulin association state, and stability in order to optimize this formulation. Here, we formulated insulin with different preservatives and stabilizing excipient concentrations using both insulin lispro and regular human insulin and assessed the insulin association states using analytical ultracentrifugation as well as formulation stability. We determined that phenoxyethanol is required to eliminate hexamers and promote a high monomer content even in a zinc-free lispro formulation. There is also a concentration dependent relationship between the concentration of polyacrylamide-based copolymer excipient and insulin stability, where a concentration greater than 0.1g/mL copolymer is required for a mostly monomeric zinc-free lispro formulation to achieve stability exceeding that of Humalog in a stressed aging assay. Further, we determined that under the formulation conditions tested zinc-free regular human insulin remains primarily hexameric and is not at this time a promising candidate for rapid-acting formulations.

Biochemical Characteristics and Healing Activity of Bromelia laciniosa Leaf Protease

Macambira plant (Bromelia laciniosa Mart. Ex Shult. & Schult. f.) is a species belonging to the family Bromeliaceae, which is distributed widely in the hinterlands of northeastern Brazil, from Bahia to Piauí states. The plants of this species contain an enzyme complex named bromelain, which has great potential for therapeutic applications. In the present study, bromelain derived from Bromelia laciniosa leaves was assessed for its thermodynamic properties and in vitro cytotoxic and healing activities. Bromelain was extracted from the leaves and partitioned using ethanol. The partially purified bromelain was then characterized to determine the optimum pH and temperature and the kinetic and thermodynamic parameters. The partially purified bromelain derived from macambira leaves exhibited its highest activity at 47°C and pH 7, when the Km and Vmax values of 2.26 µM and 2.083 U/mL, respectively. The half-life of this bromelain at 47°C was 5.78 h. At 47°C, the Gibbs's free energy, enthalpy, and entropy values reached 89, 53.54, and –0.11 kJ/mol K, respectively. In the SDS-PAGE analysis, bromelain appeared as the main protein band, with an estimated molecular mass of 33 kDa. The in vitro cytotoxic activity and wound healing effects were determined using the MTT assay and Scratch assay, respectively. These assays were performed on the non-neoplastic cell lines L-929 and PBMC and the human tumor cell lines MCF-7, HCT-116, and NCI-H292. Bromelain exhibited a positive healing activity with a change rate similar to that observed for the control. In addition, bromelain exhibited cytotoxic activity against NCI-H292 tumor cells and chemopreventive potential. These results indicate that bromelain derived from macambira leaves may be utilized as a potential ingredient in pharmaceutical formulations, for various healing and chemopreventive purposes and as an alternative to the bromelain extracted from pineapple. In addition, the biochemical characteristics of this bromelain could find application in the food industry. Future studies should, therefore, focus on deciphering the action mechanisms underlying the activities of bromelain observed in the present study to further confirm and enhance its application potential for therapeutic purposes.

Chemical Profile of Lipophilic Fractions of Different Parts of Zizyphus lotus L. by GC-MS and Evaluation of Their Antiproliferative and Antibacterial Activities.

Zizyphus lotus L. is a perennial shrub particularly used in Algerian folk medicine, but little is known concerning the lipophilic compounds in the most frequently used parts, namely, root bark, pulp, leaves and seeds, which are associated with health benefits. In this vein, the lipophilic fractions of these morphological parts of Z. lotus from Morocco were studied by gas chromatography–mass spectrometry (GC–MS), and their antiproliferative and antimicrobial activities were evaluated. GC–MS analysis allowed the identification and quantification of 99 lipophilic compounds, including fatty acids, long-chain aliphatic alcohols, pentacyclic triterpenic compounds, sterols, monoglycerides, aromatic compounds and other minor components. Lipophilic extracts of pulp, leaves and seeds were revealed to be mainly composed of fatty acids, representing 54.3–88.6% of the total compounds detected. The leaves and seeds were particularly rich in unsaturated fatty acids, namely, (9Z,12Z)-octadeca-9,12-dienoic acid (2431 mg kg−1 of dry weight) and (9Z)-octadec-9-enoic acid (6255 mg kg−1 of dry weight). In contrast, root bark contained a high content of pentacyclic triterpenic compounds, particularly betulinic acid, accounting for 9838 mg kg−1 of dry weight. Root bark extract showed promising antiproliferative activity against a triple-negative breast cancer cell line, MDA-MB-231, with a half-maximal inhibitory concentration (IC50) = 4.23 ± 0.18 µg mL−1 of extract. Leaf extract displayed interesting antimicrobial activity against Escherichia coli, methicillin-sensitive Staphylococcus aureus and Staphylococcus epidermis, presenting minimum inhibitory concentration (MIC) values from 1024 to 2048 µg mL−1 of extract. Our results demonstrate that Zizyphus lotus L. is a source of promising bioactive components, which can be exploited as natural ingredients in pharmaceutical formulations.

Sensor Applications in Analysis of Drugs and Formulations

Biosensors are currently widely used in biomedical diagnostics, as well as point-of-care assessment of therapy and disease advancement, environmental sensing, food safety, drug development, forensics, and biomedical research. Biosensors may be developed using several different approaches. Due to the growing requirement for efficient and low-cost analytical methods, biosensors have gained increasing attention for application in the quality analysis of pharmaceuticals and other pharmaceutically relevant analytes. Biosensors enable the analysis of active ingredients in pharmaceutical formulations as well as the determination of degraded products and intermediates in biological matrices. The current study discusses several types of biosensors and their applications in drug analysis and formulations.

Promising Technological and Industrial Applications of Deep Eutectic Systems.

Deep Eutectic Systems (DESs) are obtained by combining Hydrogen Bond Acceptors (HBAs) and Hydrogen Bond Donors (HBDs) in specific molar ratios. Since their first appearance in the literature in 2003, they have shown a wide range of applications, ranging from the selective extraction of biomass or metals to medicine, as well as from pollution control systems to catalytic active solvents and co-solvents. The very peculiar physical properties of DESs, such as the elevated density and viscosity, reduced conductivity, improved solvent ability and a peculiar optical behavior, can be exploited for engineering modular systems which cannot be obtained with other non-eutectic mixtures. In the present review, selected DESs research fields, as their use in materials synthesis, as solvents for volatile organic compounds, as ingredients in pharmaceutical formulations and as active solvents and cosolvents in organic synthesis, are reported and discussed in terms of application and future perspectives.

A flow analysis system integrating an optoelectronic detector for the quantitative determination of active ingredients in pharmaceutical formulations

A simple, compact and low-cost optoelectronic detector was developed for complete turbidimetric measurements under continuous flow injection analysis. The device was fabricated by integrating white snow light-emitting diodes as a light source with solar cells as a detector (WLEDs-SCs) of attenuated light that is passed through the analyte. The device is made from ten of WLEDs and three solar cells only without any filters, lenses and fibers. The device operates according to the turbidimetric principle which measures the attenuated light at 180° of the incident light. The developed detector was adapted under flow injection analysis conditions and its real analytical utility has been successfully confirmed by validation and the development of a turbidimetric method for the quantitative determination of Cyproheptadine hydrochloride (CPH) in pure and pharmaceutical formulations. The proposed method is based on forming a turbid, yellow ion-pair association complex in a closed flow system as a result of a reaction between the drug and potassium chromate which was used as a precipitation reagent. Under optimized conditions, the analysis system offers linear range 2.2–160 µg ml−1 with a limit of detection at 0.6 µg ml−1, limit of quantification 1.8 µg ml−1 and correlation coefficient of 0.9986. The intra-day and inter-day precision for two serial estimations of 30 and 60 µg ml−1 of CPH exhibited a relative standard deviation of 0.94%, 0.29 and 0.68%, 0.21% respectively. While the accuracy of the proposed method has expressed as a recovery percentage (Rec %) and error % which were between 100.45–100.67 and 0.45% to 0.67% respectively. The proposed method has been successfully applied for the CPH determination in pharmaceutical preparations with high throughput tests (60 injections per hour) with no interference effect of the common excipients.

LICENSING AND LEGALIZATION OF TRADITIONAL MEDICINE TO ENSURE PUBLIC HEALTHH QUALITY

Traditional medicine is a form of pharmaceutical ingredient which uses as medicine by the people. In public paradigm, traditional medicines is considering more secure and cheaper compare with patent medicine. The reason of such condition due to traditional medicines are made from materials of plants, animals, mineral galenic ingredients or mix of all those ingredients hereditary. In Indonesia the instrument control of traditional medicine conducted by regulation, license, supervision and legal enforcement. In practical, based on the surveillance result which conducted by BPOM found from 7.605 sample of traditional medicine, 1.867 (24.55%) of those are not fulfilled the standard because several reasons contain chemical ingredients, does not have distribution license, has long wrecked time and has unsuitable water content. This condition resulted due to the lack of ability of traditional medicine business to fulfill the requirement and obligation of license owner to conduct the medicine manufacture base on CPOTB. Keywords: traditional medicine, license, legal enforcement.

Micellar systems of poloxamer 407 and 403 for quercetin delivery

The use of poloxamers (PL) have been one of the most widely strategies used for drug delivery systems. The copolymer Poloxamer 407 is a non-ionic surfactant consisting of polioxyethylene (POE) and polioxypropylene (POP) that has many pharmaceutical applications due to its thermoreversible properties. PL are biocompatible and for this reason, are very attractive as carrier systems for administration by different routes, including topical. This work aimed the development with emphasis on physicohemical characterization of PL hydrogels for quercetin, being a useful strategy to expand the therapeutic application of these compounds as active ingredients in pharmaceutical formulations designed for topical use.1, 2 In the first stage this work was carried out to study the physico-chemical characteristics of quercetin and evelopment of formulations containing PL gels 407 (28% to 30%) and PL 403 (2%) associated to solubilizing agents polyethyleneglycol 600 (PEG 600) and propyleneglycol (PPG). Systems with higher linearity suggest that PL hydrogels were efficient for permeation across artificial membranes. Besides, thermodynamic profile was obtained from Differential Scanning Calorimetry (DSC) assays, best values of enthalpy (0.580 J / g) and Gibb's free energy (-12245.95 J.K1.mol-1) were observed for the system PL 407-PEG 600, showing that PL system present spontaneous micellization, stability and thermorreversibility for quercetin delivery.

Chemometric analysis of antioxidant properties of herbal products containing Ginkgo biloba extract

AbstractGinkgo biloba extract is a popular ingredient in pharmaceutical formulations. The level of bioactive compounds determines final antioxidant activity of an extract and its therapeutic efficiency. The aim of the project was to assess phytopharmaceuticals containing Ginkgo extracts in terms of their chemical characteristic and antioxidant activity using selected chemometric methods. The aim of the study was also to investigate whether the antioxidant activity of phytopharmaceuticals can be successfully predicted based on their chromatograms. Total polyphenol content and antioxidant activity of methanolic Ginkgo extracts were determined using a spectrophotometric technique. Comparative chemometric analysis of investigated pharmaceutical formulations was carried out using hierarchical and non-hierarchical segmentation algorithms. Samples of herbal drug products were clearly separated from samples of dietary supplements. A tree regression method was used to predict the antioxidant activity of pharmaceuticals based on the chromatographic description. The correlations between the predicted (by regression model) and experimental values of IC50 for training and testing subset were 0.960 and 0.949, respectively. The chemometric techniques, combined with instrumental analysis, could improve and simplify the quality control methods of herbal products. The antioxidant activity, predicted on the basis of chromatograms, may be one of measures of final product quality.

HPTLC methods to assay active ingredients in pharmaceutical formulations: A review of the method development and validation steps

High-performance thin-layer chromatography (HPTLC) is still increasingly finding its way in pharmaceutical analysis in some parts of the world. With the advancements in the stationary phases and the introduction of densitometers as detection equipment, the technique achieves for given applications a precision and trueness comparable to high-performance liquid chromatography (HPLC). In this review, the literature is surveyed for developed and validated HPTLC methods to assay active ingredients in pharmaceutical formulations published in the period 2005-2011. Procedures and approaches for method development, validation and quantitative assays are compared with the standard ways of conducting them. Applications of HPTLC in some other areas are also briefly highlighted.

Determination of ceftriaxone, ceftizoxime, paracetamol, and diclofenac sodium by capillary zone electrophoresis in pharmaceutical formulations and in human blood serum

A simple and fairly fast capillary zone electrophoresis method has been developed for the determination and separation of ceftriaxone, ceftizoxime, paracetamol, and diclofenac sodium in a mixture, in pharmaceutical formulations, and in blood serum. A 50 mM sodium tetraborate background electrolyte solution (pH 9.0) was found to be suitable for separation of all the drugs. An uncoated fused-silica capillary of a total length of 57 cm (effective length 50 cm) was used for separation. All the analytes were completely separated within 8.0 min at an applied voltage of 30 kV and detection was performed at 214 nm .Validation of the method was performed in terms of linearity, accuracy, precision and limit of detection, and quantification. The linearity of the calibration curves for paracetamol was 5-125 \mu g/mL, for ceftizoxime was 5-500 \mu g/mL, for diclofenac was 1-125 \mu g/mL, and for ceftriaxone was 10-1000 \mu g/mL, while LOD of the paracetamol, ceftizoxime, diclofenac, and ceftriaxone was found to be 1.0, 1.0, 0.5, and 5.0 \mu g/mL, respectively. The proposed method was applied for the determination of active ingredients in pharmaceutical formulations and active drugs in human blood serum. The recovery was found to be >= 99% with the relative standard deviation (RSD) Keywords: Capillary zone electrophoresis, paracetamol, ceftriaxone, ceftizoxime, diclofenac sodium

Simultaneous separation of epinephrine and norepinephrine enantiomers by EKC: Application to the analysis of pharmaceutical formulations

A new analytical methodology using EKC with a CD as chiral selector was developed enabling the simultaneous enantiomeric separation of two chiral neurotransmitters (epinephrine and norepinephrine). Different experimental conditions (type and concentration of the chiral selector, nature and concentration of buffer and separation voltage) were optimized. The use of 10 mM carboxyethyl-beta-CD in 100 mM Tris-phosphate buffer (pH 5.0) with an applied voltage of 30 kV and a temperature of 15 degrees C allowed the separation of epinephrine and norepinephrine enantiomers with high chiral resolution (Rs>2.5). Moreover, after evaluating the analytical characteristics of the method, it was applied to the quantitation and purity testing of L-epinephrine as active pharmaceutical ingredient in pharmaceutical formulations where D-epinephrine and L/D-norepinephrine can be present as impurities.

A New HPLC Method for the Simultaneous Determination of Acetaminophen, Phenylephrine, Dextromethorphan and Chlorpheniramine in Pharmaceutical Formulations

Acetaminophen, phenylephrine, dextromethorphan, and chlorpheniramine are frequently associated in pharmaceutical formulations against the common cold. A new high performance liquid chromatography (HPLC) method has been developed for the simultaneous determination of these active pharmaceutical ingredients in pharmaceutical formulations. The separation and quantitation were achieved on a 25 cm underivatized silica column using a mobile phase of methanol: water (containing 6.0 g of ammonium acetate and 10 ml of triethylamine per liter, pH adjusted to 5.0 with orthophosphoric acid), 95:5%(v/v). Detection was carried out using a variable wavelength UV-vis detector at 254 nm for acetaminophen, at 220 nm for phenylephrine, and at 227 nm for dextromethorphan and chlorpheniramine. The method showed linearity for the acetaminophen, phenylephrine, dextromethorphan, and chlorpheniramine in the 162.5–650, 2.5–10, 7.5–30, and 1–4 µg/ml ranges, respectively. The intraday and interday RSDs ranged from 0.92 to 1.52%, 1.00 to 1.76%, 1.21 to 1.74% and 1.26 to 1.80% for the acetaminophen, phenylephrine, dextromethorphan, and chlorpheniramine, respectively. Compounds were eluted in a run time of less than 12 min.

Verminoside- and verbascoside-induced genotoxicity on human lymphocytes: Involvement of PARP-1 and p53 proteins

Verminoside and verbascoside are natural compounds present in plants used in traditional medicine. They exhibit several biological activities including anti-inflammatory, anti-bacterial and anti-tumor properties. The potential applications of these compounds as ingredients in pharmaceutical formulations and cosmetics prompted us to investigate on cytotoxic and genotoxic activity of verminoside and verbascoside on human lymphocytes using genetic toxicity assays recommended in preclinical studies by the US Food and Drug Administration (FDA). We analyzed chromosome aberrations (CAs) and sister chromatid exchanges (SCEs) as well as the mitotic index (MI) and cell viability after the treatments with verminoside and verbascoside. This report is the first to clearly demonstrate a significant increase of structural CAs and SCEs on normal human lymphocytes associated with a reduction of the MI in both verminoside- and verbascoside-treated cells. Moreover, we observed enhanced protein expression levels of PARP-1 and p53 that are key regulatory proteins involved in cell proliferation and DNA repair. Interestingly, mass spectrometric analysis of the compounds in the culture supernatants also showed that verminoside remained unchanged during the culture period while verbascoside was hydrolyzed to its derivative, caffeic acid and the last one seems to be responsible for the observed biological activity.

Determination of benzhexol hydrochloride by capillary zone electrophoresis with an end-column electrochemiluminescence detection

A capillary zone electrophoresis with end-column electrochemiluminescence (ECL) detector was described for the determination of benzhexol hydrochloride. The detection was based on the tris(2,2′-bypyridine)ruthenium(II) [Ru(bpy) 3 2+] ECL reaction with the analyte. Electrophoresis was performed using a 25 μm i.d. uncoated capillary. 10 mM sodium phosphate buffer (pH=8.0) was used as the running buffer. The solution in the detection cell was 80 mM sodium phosphate (pH=8.0) and 5 mM Ru(bpy) 3 2+. A linear calibration curve of three-orders of magnitude was obtained (with a correlation coefficient of >0.999) from 1.0×10 −8 to 1.0×10 −5 M and the limit of detection was 6.7×10 −9 M ( S/ N=3). This just provides an easy and sensitive method to determine the active ingredient in pharmaceutical formulations.

Validated HPLC determination of 2-[(dimethylamino)methyl]cyclohexanone, an impurity in Tramadol, using a precolumn derivatisation reaction with 2,4-dinitrophenylhydrazine

A new method for the determination of 2-[(dimethylamino)methyl]cyclohexanone (DAMC) in Tramadol (as active substance or active ingredient in pharmaceutical formulations) is described. The method is based on the derivatisation of 2-[(dimethylamino)methyl]cyclohexanone with 2,4-dinitrophenylhydrazine (2,4-DNPH) in acidic conditions followed by a reversed-phase liquid chromatographic separation with UV detection. The method is simple, selective, quantitative and allows the determination of 2-[(dimethylamino)methyl]cyclohexanone at the low ppm level. The proposed method was validated with respect to selectivity, precision, linearity, accuracy and robustness.