Journal: Forensic Sciences Research

Article Title: Determination of multiple drugs of abuse in human urine using dispersive liquid–liquid microextraction and capillary electrophoresis with PDA detection

doi: 10.1080/20961790.2021.1986771

Figure Lengend Snippet: Enrichment recoveries obtained with the different extraction solvents evaluated for the extraction of drugs by dispersive liquid–liquid microextraction (DLLME).

Article Snippet: 6-Monoacetylmorphine (6-MAM) hydrochloride, morphine hydrochloride, codeine phosphate, methamphetamine hydrochloride, amphetamine hydrochloride, 3,4-methylenedioxymethamphetamine (MDMA) hydrochloride, 3,4-methylenedioxyamphetamine (MDA) hydrochloride and ketamine hydrochloride were purchased from Cerilliant (Merck KGaA, Darmstadt, Germany).

Techniques:

Journal: Forensic Sciences Research

Article Title: Determination of multiple drugs of abuse in human urine using dispersive liquid–liquid microextraction and capillary electrophoresis with PDA detection

doi: 10.1080/20961790.2021.1986771

Figure Lengend Snippet: Enrichment recoveries obtained with the different dispersive solvents evaluated for the extraction of drugs by dispersive liquid–liquid microextraction (DLLME).

Article Snippet: 6-Monoacetylmorphine (6-MAM) hydrochloride, morphine hydrochloride, codeine phosphate, methamphetamine hydrochloride, amphetamine hydrochloride, 3,4-methylenedioxymethamphetamine (MDMA) hydrochloride, 3,4-methylenedioxyamphetamine (MDA) hydrochloride and ketamine hydrochloride were purchased from Cerilliant (Merck KGaA, Darmstadt, Germany).

Techniques:

Journal: Forensic Sciences Research

Article Title: Determination of multiple drugs of abuse in human urine using dispersive liquid–liquid microextraction and capillary electrophoresis with PDA detection

doi: 10.1080/20961790.2021.1986771

Figure Lengend Snippet: The performance of the proposed methods in spiked urine samples.

Article Snippet: 6-Monoacetylmorphine (6-MAM) hydrochloride, morphine hydrochloride, codeine phosphate, methamphetamine hydrochloride, amphetamine hydrochloride, 3,4-methylenedioxymethamphetamine (MDMA) hydrochloride, 3,4-methylenedioxyamphetamine (MDA) hydrochloride and ketamine hydrochloride were purchased from Cerilliant (Merck KGaA, Darmstadt, Germany).

Techniques:

Journal: Forensic Sciences Research

Article Title: Determination of multiple drugs of abuse in human urine using dispersive liquid–liquid microextraction and capillary electrophoresis with PDA detection

doi: 10.1080/20961790.2021.1986771

Figure Lengend Snippet: Electropherograms obtained for the spiked biological samples before (a) and after (b) dispersive liquid–liquid microextraction (DLLME) extraction under the optimum conditions. Extraction conditions: dispersive solvent, 0.5 mL isopropyl alcohol; extraction solvent, 41.0 µL CHCl 3 ; room temperature; pH of sample, 9.0; analytes concentration spiked, urine: 50 ng/mL; internal standard: 3 µg/mL lidocaine. Peak identification: (IS) lidocaine, (1) AM: amphetamine, (2) MA: methamphetamine, (3) MDA: methy­lenedioxyamphetamine, (4) MDMA: methylenedioxymethamphetamine, (5) ketamine, (6) codeine, (7) morphine, (8) 6-MAM: 6-monoacetylmorphine.

Article Snippet: 6-Monoacetylmorphine (6-MAM) hydrochloride, morphine hydrochloride, codeine phosphate, methamphetamine hydrochloride, amphetamine hydrochloride, 3,4-methylenedioxymethamphetamine (MDMA) hydrochloride, 3,4-methylenedioxyamphetamine (MDA) hydrochloride and ketamine hydrochloride were purchased from Cerilliant (Merck KGaA, Darmstadt, Germany).

Techniques: Concentration Assay

Journal: Forensic Sciences Research

Article Title: Determination of multiple drugs of abuse in human urine using dispersive liquid–liquid microextraction and capillary electrophoresis with PDA detection

doi: 10.1080/20961790.2021.1986771

Figure Lengend Snippet: Electropherograms obtained for the real biological samples from the drug abuser before (a) and after (b) dispersive liquid–liquid microextraction (DLLME) extraction under the optimum conditions. Extraction conditions: dispersive solvent, 0.5 mL isopropyl alcohol; extraction solvent, 41.0 μL CHCl 3 ; room temperature; pH of sample, 9.0; analytes concentration spiked, urine: 50 ng/mL; internal standard: 3 μg/mL lidocaine. Peak identification: (IS) lidocaine, (1) AM: amphetamine, 42.1 ng/mL, (2) MA: methamphetamine, 102.6 ng/mL, (5) ketamine, 84.7 ng/mL.

Article Snippet: 6-Monoacetylmorphine (6-MAM) hydrochloride, morphine hydrochloride, codeine phosphate, methamphetamine hydrochloride, amphetamine hydrochloride, 3,4-methylenedioxymethamphetamine (MDMA) hydrochloride, 3,4-methylenedioxyamphetamine (MDA) hydrochloride and ketamine hydrochloride were purchased from Cerilliant (Merck KGaA, Darmstadt, Germany).

Techniques: Concentration Assay

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Schemes for Experiments and post hoc analyses. (A) Scheme of the ketamine injection paradigm for Experiment 1 during development of LID (daily L-DOPA injections). FAS = Forelimb adjusting steps test. (B) Scheme of the injection paradigm in PD rats for Experiment 2 . AR = amphetamine-rotation test; RR = RotaRod test. (C) Scheme of the injection paradigm for Experiment 3 during development of LID (daily L-DOPA injections). (D) Verification of unilateral 6-OHDA lesion and evaluation of striatal dopamine (DA) levels after ketamine in the rats from the study shown in (A). Electrochemical detection of striatal DA content (mean ± SEM) is reduced by>95% in the lesioned side. Striatal DA content was unchanged by a 10-h-treatment of either ketamine (K; n =9), R -ketamine ( R -K; n =9) vs. vehicle (V; n = 9) 1-h before rats were euthanized, showing no effect on overall striatal DA levels by ketamine or R -ketamine-treatment compared to vehicle in either the lesioned (Lx) or the intact hemisphere. (E) Verification of unilateral 6-OHDA lesion from the study shown in (B) using semi-quantitative TH western analysis in striatal tissue plotting % loss (mean ± SEM) in Lx vs. intact (In) hemisphere ( n =9). Two-tailed t -test, *** p < .001. (F) Verification of unilateral 6-OHDA lesions in the rats from the study depicted in (C). The graph shows the quantification of the TH-ir plotting the % loss (mean ± SEM) in the Lx vs. intact SN hemispheres ( n =10/group; V =vehicle, K =ketamine, K +A = ketamine+ ANA-12). (G) Example photomicrograph of a SN in Experiment 3 shows the unilateral reduction in TH-ir post-lesion. Two-way ANOVAs, Bonferroni post hoc tests, *** p < .001. (H) Verification of unilateral 6-OHDA lesion from the ANA-12-only control study, the negative control for Experiment 3 , using semi-quantitative TH western analysis in striatal tissue, plotting % loss (mean ± SEM) in Lx vs. intact hemisphere ( n = 10). Two-tailed t -test, *** p < .001.

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection, Western Blot, Two Tailed Test, Negative Control

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Low-dose racemic ketamine treatment once a week attenuates the development of LID in the preclinical model. In Experiment 1 6-OHDA-lesioned PD rats were injected daily with L-DOPA (days 0–13: 6 mg/kg; days 14–28: 12 mg/kg; i.p. ) to induce dyskinesia and tested for LAO-AIMs twice a week for 3 h by blinded investigators. (A) The mean LAO AIMs scores±SEM are plotted showing a 50% reduction after racemic low-dose ketamine treatments (K) when compared to the vehicle group (V) and a group treated with R -ketamine ( R -K), to test for contribution of the stereospecific ketamine isomer. The blue arrows point to the days of the 10-h racemic ketamine (20 mg/kg; i.p. ), R -ketamine (10 mg/kg; i.p. ) or vehicle treatment paradigm; n = 9 per group, * p < .05, ** p < .01, Kruskal-Wallis test with Dunn’s multiple comparisons post hoc tests. (B) Example time course of the LAO-AIMs data showed in (A) for day 11. (C) Example time course of the LAO-AIMs data showed in (A) for day 25. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: LAO-AIMs scores from Experiment 1 separated by sub-type of dyskinesia. Low-dose racemic ketamine treatment once a week attenuates the development of (A) limb (B) axial (C) orolingual AIMs in the preclinical model. 6-OHDA-lesioned PD rats were injected daily with L-DOPA (days 0–13: 6 mg/kg; days 14–28: 12 mg/kg; i.p. ) to induce dyskinesia and tested for LAO AIMs twice a week for 3 h by blinded investigators. The mean LAO AIMs scores ± SEM are plotted showing a 50% reduction after racemic low-dose ketamine treatments (K) when compared to the vehicle group (V) and a group treated with R -ketamine ( R -K), to test for contribution of the stereospecific ketamine isomer. The blue arrows point to the days of the 10-h racemic ketamine (20 mg/kg; i.p. ), R -ketamine (10 mg/kg; i.p. ) or vehicle treatment paradigm; n = 9 per group, * p < .05, ** p < .01, Kruskal-Wallis tests with Dunn’s post hoc tests. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Ketamine does not interfere with the anti-PD effect of L-DOPA and reduces PD-motor behavior post-6-OHDA-lesion (Post-Lx) by itself. (A) Mean % contralateral/ipsilateral ratios of steps ± SEM using the FAS test paradigm in the LID cohort of Experiment 1 , are plotted after normalization to pre-lesion (Pre-Lx) indicate a significant anti-PD effect of ketamine * p < .05; repeated measures ANOVA. Ketamine does also not interfere with the anti-PD effect of L-DOPA, and a significant increase of stepping contralateral to the lesioned side after either L-DOPA alone or L-DOPA + Ketamine vs. all Post-Lx time points is seen: *** p < .001; one-way ANOVA on data prior to normalization, Tukey-Kramer corrected post hoc tests; n = 9 per group. (B) In Experiment 2 we tested ketamine treatment in a separate cohort of hemi-parkinsonian 6-OHDA-lesioned rats and used the RotaRod test to evaluate the deficit. The graph shows the mean latency to fall ± SEM, normalized to pre-lesion baseline (Pre-Lx). Post-lesion (Post-Lx) the latency to fall was reduced by 50% in these PD animals. This motor deficit was reversed by ketamine treatment (blue bars), already at the 1st injection, and the animals performed as good as at baseline. One-way ANOVA, with Tukey-Kramer corrected post hoc tests, on raw data before normalization. n = 9, * p < .05, ** p < .01. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Ketamine’s long-term anti-dyskinetic activity was driven by BDNF signaling. (A) The mean LAO-AIMs scores ± SEM of Experiment 3 are plotted. The sustained anti-dyskinetic effect of low-dose ketamine is reduced by blocking the BDNF receptor TrkB, with co-injection of the TrkB antagonist ANA-12 (0.5 mg/kg; i.p. ) with ketamine (K + A). The blue arrows point to the days of the 10-h racemic (K) ketamine (20 mg/kg; i.p. ), or vehicle treatment paradigm (V). ANA-12 co-injection (green bars) did reduce the sustained anti-dyskinetic effect seen in ketamine-only injected LID (blue bars) leading to LAO AIMs comparable to those of the vehicle group (grey bars), indicating an involvement of BDNF in the sustained anti-dyskinetic effects of ketamine. n = 10 per group, * p < .05, ** p < .01, ANOVAs, Tukey-Kramer corrected post hoc tests. (B) Example time course of the LAO-AIMs for day 14 showed in (A). (C) A control study using 10-h ANA-12-only treatments on days 0 and 7 of daily L-DOPA-treatment (6 mg/kg; i.p .) verified that, while systemic TrkB antagonism does block the ketamine effect, it does not change development of LID in this model, and serves as an important negative control for the data shown in (A). The graph depicts the mean LAO-AIMs scores ± SEM from the vehicle control groups in Experiment 1 (V- E1 ; n = 9) and Experiment 3 (V- E3 ; n = 10), as well as the ANA-12-only control study (ANA-12; n = 10). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Activity Assay, Blocking Assay, Injection, Negative Control

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: LAO-AIMs scores from Experiment 3 separated by sub-type of dyskinesia. Low-dose ketamine treatment (blue arrows) once a week attenuates the development of individual AIMs scores. 6-OHDA-lesioned PD rats were injected daily with L-DOPA (days 0–14: 6 mg/kg; i.p. ) to induce dyskinesia and tested for LAO-AIMs twice a week for 3 h by blinded investigators. The anti-dyskinetic effect of low-dose ketamine (K, blue bars) reduced the individual (A) limb (B) axial (C) and orolingual AIMs (mean ± SEM) scores, compared to the vehicle (V, grey bars) group and a group treated with the TrkB antagonist, ANA-12 (K + A, green bars). The blue arrows point to the days of the 10-h racemic ketamine (20 mg/kg; i.p. ), R -ketamine (10 mg/kg; i.p. ) n = 10 per group, *p < .05, **p < .01, Kruskal-Wallis tests with Dunn’s post hoc tests. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection