Greenness assessment of analytical methods for determination of cannabinoids in oils using NEMI, Analytical Eco-Scale, AGREE and GAPI.

The cannabis plant is being increasingly researched due to its numerous therapeutic properties leading to the need for analytical techniques to assess substances present in extracts of the cannabis plant in carrier oils, such as medium chain triglycerides (MCT) oil. Awareness of the environmental impact of activities related to analysis led to the development of greenness assessment metrics. This study aimed to assess the environmental impact of analytical techniques applied in the analysis of cannabinoids in oil using Green Analytical Chemistry metrics. The first phase of the study consisted of a systematic literature review to identify high performance liquid chromatography and ultra high performance liquid chromatographic methods of analysis for cannabinoids in oil. In the second phase, the identified methods were assessed using the National Environmental Method Index (NEMI), Analytical Eco-scale, Analytical Greenness Calculator (AGREE) and Green Analytical Procedure Index (GAPI). Out of 124 identified studies, 8 were considered for the comparative analysis. The identified analytical methods consisted of high performance liquid chromatography (HPLC) using high resolution MS (n = 1), DAD (n = 2), UV (n = 1), UV and MS (n = 2) and MS/MS (n = 2) as detectors. When the analytical methods were assessed using the Analytical Eco-Scale, 7 out of 8 methods achieved a score ranging between 50 and 73, categorising them as acceptable green methods of analysis. One method achieved a total score of 80, categorising the method as an excellent green analysis. The application of Green Analytical Chemistry and respective metrics during the development of analytical methods contributes towards a reduction in the environmental footprint which results from related activities.

Challenges of Extracting and Determining Cannabinoids in Different Matrices.

Introduction: Accurate and precise analysis of cannabinoids is important for elucidating their therapeutic potential and developing therapies, which are targeted toward different medical conditions. A wide range of cannabis products are present on the market and are available in different dosage forms, including dried flowers, extracts, and consumables. The aim of this article is to provide an updated narrative review of literature on challenges of analyzing cannabinoids in plant material, oils, and edibles. Method: Literature search was conducted to identify sample preparation and analytical techniques for determination of cannabinoids in plant material, oils, and edibles and associated challenges. Results: Challenges related to determination of cannabinoids in plant material include matrix complexity, co-extraction of unwanted compounds during sample preparation, and differences in matrix composition between calibration standards and sample extracts. During analysis of cannabinoids in oil, the unique properties of carrier oils need to be taken into consideration. Analysis of cannabinoids in edibles can be considered to be challenging due to the wide range of matrix types that are available on the market, rendering analysis resource-intensive, time-consuming, and impractical. Discussion: Analysis of cannabinoids in plant material, oils, and edibles requires a multifaceted approach that includes regulatory guidance, method development, and technological innovation. In the face of an evolving analytical landscape where novel cannabinoids are being identified and require determination, there is a need for the development and validation of standardized accurate and precise analytical methods, which are specifically tailored for each matrix.

HPLC-UV Method Development and Validation to Monitor Difluprednate Synthesis.

During the synthesis of active pharmaceutical ingredients (APIs) there is a need for the development and validation of a simple and rapid high performance liquid chromatography (HPLC) method for the determination and quantification of the synthesized product and related by-products. An HPLC method gives a better understanding of how a synthesis is proceeding. A rapid and easy to use HPLC-UV (ultraviolet) method for the determination of difluprednate and monitoring of impurities generated during synthesis was developed and validated. A Shimadzu VP Series HPLC equipped with a LabSolutions software and UV detector set at 240 nm was used for analysis. The mobile phase consisted of phosphate buffer (pH 6) and acetonitrile 50:50 (v/v) and was eluted at a flow rate of 1.2 mL/min. Separation took place on a reversed-phase Kinetex C18 column (150 × 4.60 mm; 5 µm i.d.). Column temperature was set at 40°C. The developed method was found to have good linearity and acceptable accuracy and precision. The developed method may be effectively applied to determine products and by-products formed during synthetic reactions of steroids and to calculate the yield of the products obtained during each step of the synthesis.

QT shortening: a proarrhythmic safety surrogate measure or an inappropriate indicator of it?

OBJECTIVE: To examine whether QT interval shortening is an overlooked adverse event as compared to QT prolongation through a review of preclinical, clinical and post-marketing adverse event data available to the regulator for centrally and nationally authorized medicinal products. METHODS: Potential safety signals of QT shortening related to authorized medicinal products were detected from Eudravigilance using proportional reporting ratios. Active substances identified as having unexpected signals of QT shortening were assessed in depth using the Bradford-Hill criteria for causation. Preclinical, clinical and adverse event data related to each active substance was used in the assessments. Post marketing adverse event cases were reviewed for imputability using the French method. RESULTS: 80 adverse event cases of electrocardiogram QT shortening were detected from 13 different active substances which included antipsychotics and antiepileptics (Clozapine, Ziprasidone, Quetiapine, Olanzapine, Carbamazepine), cardiovascular drugs (Atenolol, Digoxin, Ramipril, Simvastatin), anti-inflammatories and analgesics (Ibuprofen, Paracetamol) and other substances Calcium Carbonate (Mineral Supplement/Antacid) and Fingolimod (Immunosuppressant). By comparison 404 active substances were found have a potential safety signal of Electrocardiogram QT prolongation. Following in depth review none of the 13 active substances identified were found to be clearly associated with QT shortening using the minimum level of evidence for regulatory action. In the preclinical data reviewed we observed cases of morphological changes to the action potential (AP) where the Action Potential Duration at 90% (APD90) was not affected. CONCLUSIONS: From a regulatory perspective one cannot refute the possibility of a clinically relevant risk from QT shortening through the current testing requirements. Lack of further investigations into any potential morphological changes to the AP, or APD90 shortening beyond a specified threshold in our opinion does not fully exclude the possibility of proarrhythmic effects of active substances.

Treatment of Leber's hereditary optic neuropathy: An overview of recent developments.

Leber's hereditary optic neuropathy (LHON) is a rare, maternally-inherited optic neuropathy caused by mitochondrial DNA point mutations and which can cause blindness. Currently, Raxone (idebenone) is the only available medicinal product authorised to treat LHON within the European Union and LHON remains an unmet medical need. The aim of this article was to summarise interventional clinical trials published over the past 5 years (between 2014 and 2019) with the primary purpose of treating LHON. Therapeutic approaches discussed include modulating agents of the mitochondrial electron transport chain such as Raxone, cysteamine bitartrate and KH176, inhibitors of apoptosis such as elamipretide, gene therapy medicinal products such as GS010 and scAAV2P1ND4 and retinal tissue regeneration medicinal products such as bone marrow-derived stem cells.

Practical liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of amitriptyline, nortriptyline and their hydroxy metabolites in human serum.

Amitriptyline (AMI) has been in use for decades in treating depression and more recently for the management of neuropathic pain. A highly sensitive and specific LC-tandem mass spectrometry method was developed for simultaneous determination of AMI, its active metabolite nortriptyline (NOR) and their hydroxy-metabolites in human serum, using deuterated AMI and NOR as internal standards. The isobaric E-10-hydroxyamitriptyline (E-OH AMI), Z-10-hydroxyamitriptyline (Z-OH AMI), E-10-hydroxynortriptyline (E-OH NOR) and Z-10-hydroxynortriptyline (Z-OH NOR), together with their parent compounds, were separated on an ACE C18 column using a simple protein precipitation method, followed by dilution and analysis using positive electrospray ionisation with multiple reaction monitoring. The total run time was 6 min with elution of E-OH AMI, E-OH NOR, Z-OH AMI, Z-OH NOR, AMI (+ deuterated AMI) and NOR (+ deuterated NOR) at 1.21, 1.28, 1.66, 1.71, 2.50 and 2.59 min, respectively. The method was validated in human serum with a lower limit of quantitation of 0.5 ng/mL for all analytes. A linear response function was established for the range of concentrations 0.5-400 ng/mL (r2 > .999). The practical assay was applied on samples from patients on AMI, genotyped for CYP2C19 and CYP2D6, to understand the influence of metaboliser status and concomitant medication on therapeutic drug monitoring.