Drug Samples Research Articles

Disposable and low-cost lab-made screen-printed electrodes for voltammetric determination of L-dopa

This work presents a feasible process for preparation screen-printed electrode (SPE) using a lab-made conductive ink based on cellulose acetate (CAc) and graphite powder (GP) printed on polyvinyl chloride (PVC) sheet. The CAc present in the composition of this SPE is a binding agent for conductive inks with GP, which are relatively low-cost and easily obtainable materials, compared to other similar commercial sensors. The SPE was evaluated through cyclic voltammetry (CV) using an equimolar [Fe(CN)6]3-/4− solution as a redox probe to find an adequate ink composition. Electrode surface was characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). To show the analytical potential of SPE proposed it was used for Levodopa (L-dopa) determination. L-dopa is a drug frequently used to reduce the symptoms of Parkinson's disease. The GP-CAc/PVC electrode was evaluated for the determination of L-dopa under square wave voltammetric (SWV) conditions. The electrode presented a linear dynamic range (LDR) from 8.00 to 100 µmol L−1 and a limit of detection (LOD) of 0.06 µmol L−1. As a proof-of-concept, the proposed electrode was used to quantify L-dopa in drug samples as an alternative tool for quality control tests, being applied to two commercial drugs. The electrode construction strategy proved to be reproducible with an analytical performance comparable to other similar electrodes reported in the literature.

Bioanalytical Method Development and Validation for the Determination of Favipiravir in Spiked Human Plasma by using RP-HPLC

A precise, simple and reproducible reverse phase liquid chromatography (RP-HPLC) method was developed and validated for determination of Favipiravir by using Carbamazepine as internal standard in spiked human plasma. A chromatographic separation was accomplished with Cromasil C18 (250mm x 4.6ID, Particle size: 5 micron) column using mobile phase consists of methanol: water in the ratio (35:65, %v/v), at pH 3.0 with binary gradient system-maintained flow rate at 0.8ml/min. The detection wavelength of drug sample was at 225 nm. Extraction was done by using ethyl acetate as extracting solvent. The retention time of Favipiravir was found to be 6.62 min. The method was found to be linear in the concentration range of 0.2-3.2 µg/ml. Limit of quantitation (LOQ) value was found to be 0.72. The intra- and inter day precision and accuracy lies within the specified range. The recovery studies were found to be in the range of 97.6 to 100.2%. %Relative standard deviation (RSD) was found to be in the range of 0.07-2.80%. All parameters were found to be validated from spiked human plasma. The proposed RP-HPLC method is highly accurate and rapid for the determination of favipiravir in human plasma and can be applied for pharmacokinetic studies and Therapeutic drug monitoring.

In vitro Antiradical activity and Hepatoprotective potential of Herbal extract from Medicinal plants of Rosaceae family

The experimental study was designed to evaluate the in vitro antioxidant activity and hepatoprotective potential of ethanolic fruit extract of Pyrus pyrifolia (Burm.f.) Nakai. and Prunus avium Linn. in carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. Pharmacognostic evaluation of fruits and their extracts was carried out according to the WHO quality control of herbal drugs. The antiradical activity of ethanol extracts of P. pyrifolia (Burm.f.) Nakai and P. avium L. was determined by using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2) and nitric oxide free radical scavenging assay. The Wistar albino rats were randomly divided into seven equal groups (n = 5). Group I was treated as normal control. Group II received 0.2% carbon tetrachloride (CCL4) in olive oil (8 mL/kg, i.p) on 14th day; Groups III as standard control contained silymarin (100 mg/kg, p.o) as standard drug once daily for 14 days. Group IV and V were given P. pyrifolia (Burm.f.) Nakai extract (200 and 400 mg/kg, p.o. in olive oil) for fourteen days; Group VI and VII received P. avium Linn. extract (200 and 400 mg/kg p.o. in olive oil) for fourteen days. Group III to VII animals were treated with 0.2% CCl4 in olive oil (8mL/kg, i.p) on the 14th day after the administration (1 hour) of the standard drug and test samples. Histopathological analysis and measurement of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein (TP), total albumin (TA) and total bilirubin (TB) were performed. Ethanol extract of P.avium L. demonstrated stronger antioxidant and higher total phenolic as well as total flavonoid content than P. pyrifolia (Burm.f.) Nakai. Both the extracts EEPP and EEPA (200 and 400 mg/kg) significantly normalized the CCl4-elevated levels of ALT, AST and ALP. P. pyrifolia (Burm.f.) Nakai and P. avium L. are the edible seasonal fruits that possessed an appreciable amount of phenolic and flavonoid components and could provide considerable protection against CCl4 hepatotoxicity in rodents that may be related to its antioxidant properties and also useful in preventing several degenerative and life threatening diseases.

Outcomes of a 12-week ecologically valid observational study of first treatment with methylphenidate in a representative clinical sample of drug naïve children with ADHD.

Randomized placebo-controlled trials have reported efficacy of methylphenidate (MPH) for Attention-deficit/hyperactivity disorder (ADHD); however, selection biases due to strict entry criteria may limit the generalizability of the findings. Few ecologically valid studies have investigated effectiveness of MPH in representative clinical populations of children. This independently funded study aims to describe treatment responses and their predictors during the first 12 weeks of MPH treatment using repeated measurements of symptoms and adverse reactions (ARs) to treatment in 207 children recently diagnosed with ADHD. The children were consecutively included from the Child and Adolescent Mental Health Centre, Mental Health Services, The Capital Region of Denmark. The children (mean age, 9.6 years [range 7-12], 75.4% males) were titrated with MPH, based on weekly assessments of symptoms (18-item ADHD-rating scale scores, ADHD-RS-C) and ARs. At study-end 187 (90.8%) children reached a mean end-dose of 1.0 mg/kg/day. A normalisation/borderline normalisation on ADHD-RS-C was achieved for 168 (81.2%) children on the Inattention and/or the Hyperactivity-Impulsivity subscale in week 12, and 31 (15.0%) children were nonresponders, which was defined as absence of normalisation/borderline normalisation (n = 19) or discontinuation due to ARs (n = 12), and eight (3.8%) children dropped out from follow-up. Nonresponders were characterised by more severe symptoms of Hyperactivity-Impulsivity and global impairment before the treatment. ARs were few; the most prominent were appetite reduction and weight loss. A decrease in AR-like symptoms during the treatment period questions the validity of currently available standard instruments designed to measure ARs of MPH. This ecologically valid observational study supports prior randomized placebo-controlled trials; 81.2% of the children responded favourably in multiple domains with few harmful effects to carefully titrated MPH. Clinical trial registration: ClinicalTrials.gov with registration number NCT04366609.

Obtaining and research of pharmaceutical properties of antiemetics in a form of powder for inhalation

Samples of antiemetic drugs (ondansetronum, palonosetronum, metoclopramidum) in the form of powder for inhalation have been developed by the method of spray drying. The granulometric composition, hygroscopicity and aerodynamic distribution of aerosol particles of the drugs have been investigated. The dosage form of the powder for inhalation of antiemetics (ondansetronum and palonosetronum) in terms of its particle size distribution, hygroscopicity and content of the agent corresponds to those for inhalation using dry powder inhalers. In the study of the phase-dispersed composition of aerosol, ondansetronum and palonosetronum in the dosage form of powder for inhalation as part of the HandiHaler inhaler (at a flow rate of 60 l/min) showed high rates of the released dose up to 7276%, respirable particle fraction (up to 5 m) up to 5456% and a mass median particle size of about 3 microns. Obtaining the inhaled form of metoclopramide requires optimization of the production method for receiving the product with acceptable pharmaceutical properties.

A generic gas chromatography method for determination of residual solvents in PET radiopharmaceuticals

A novel gas chromatography (GC) method for quantitation of volatile organic compounds (VOCs) in 18F- and 11C-radiopharmaceuticals listed in the European Pharmacopoeia (Ph. Eur.) was proposed. Optimized chromatographic parameters were used for separation of ethanol, acetone, acetonitrile, tetrahydrofuran (THF), dibromomethane (DBM), 2-dimethylaminoethanol (deanol), N,N-dimethylformamide (DMF) and dimethyl sulfoxide (DMSO) which could be detected in radioactive drug samples. The calculated peak resolutions (RS) were higher than 2.0 at ethanol concentration of up to 11 m/m%. Reproducible results could be obtained using base deactivated fused silica wool as packing material of inlet liner. Validation parameters showed excellent linearity (r2 ≥0.9998) in the range from 10 to at least 120% of concentration limit of solvents. The accuracy was determined as recovery of concentrations which ranged from 99.3% to 103.8%. Additionally, the relative standard deviation (RSD) of each solvent for inter-day and intra-day precision were in the range of 0.5–4.2% and 0.4–4.4%, respectively. The limit of quantitation (LOQ) for ethanol, acetone, acetonitrile, THF, DBM, deanol, DMF and DMSO was 0.48, 0.42, 0.43, 0.46, 4.35, 0.73, 0.68 and 0.50 mg/L, respectively. The developed procedure was successively applied for quantitation of ethanol, acetone, acetonitrile and deanol in radioactive drug samples of [11C]methionine, [11C]choline, 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) and O-(2-[18F]fluoroethyl)-L-tyrosine ([18F]FET). The proposed GC method applying flame ionization detection (FID) could be adapted in routine quality control of most frequently used positron emission tomography (PET) radiopharmaceuticals to perform the determination of residual solvents with analysis time of 12 min

Analytical characterization of biomarkers in an optimized novel antidiabetic polyherbal formulation using high-performance thin-layer chromatography and liquid chromatography with tandem mass spectrometry

Background Diabetes mellitus is a chronic health issue that requires novel approaches to treatment and a multimodal approach to prevention. In the treatment of diabetes, a polyherbal formulation is the finest alternative medicine. A polyherbal formulation was developed in-house and evaluated for its antidiabetic potential on streptozotocin-induced diabetes rat. The same extract was now characterized analytically utilizing a variety of methods. Objective The goal of this study was to quantify the biomarkers in a novel antidiabetic polyherbal formulation made in-house with Cinnamonum zeylanicium bark, Eugenia jambolana seeds, Vinca rosea whole plant, and Gymnema sylvestre (GS) leaves, using high-performance thin-layer chromatography (HPTLC) and liquid chromatography with tandem mass spectrometry (LC–MS/MS). Materials and methods Cinnamaldehyde (CIN), gallic acid (GLA), vincristine (VC), vinblastine (VB), and gymnemic acid (GYA) were identified as bioactive components of polyherbal formulation hydroalcoholic extract utilizing HPTLC and LC–MS/MS. Acetonitrile, methanol, and 0.1 percent formic acid was used as mobile phase, chromatographic separation was accomplished in 30 min using a gradient system and a SUNFIRE C18, 250×4.6, 5-µm analytical column with a flow rate of 1.0 ml/min in LC–MS/MS research. Scanned in a positive mode with a scan speed of 100–2000 AMU/s over a mass range of 20–1974 Da. The electron-spray ionization mode was used, with a source temperature of 150°C and a desolvation temperature of 350°C. The HPTLC separation was performed using ethyl acetate/acetonitrile/water/formic acid/N-dimethyl formamide 5.5 : 2.5 : 0.5 : 1.0 : 0.5 (v/v) as the mobile phase on precoated silica gel 60 GF254 plates. At room temperature, the plates were developed to a distance of 9.0 cm. CIN, GLA, VC, VB, and GYA plates were scanned and measured at wavelengths of maximum absorption of 259, 287, 342, 355, and 387 nm, respectively. Band size, chamber-saturation duration, migration of the solvent front, slit width, and other experimental parameters were carefully examined, and the optimized chromatographic conditions were chosen. Results LC–MS analysis of the hydroalcoholic extract of the polyherbal formulation revealed the presence of all the five bioactive chemical constituents, CIN, GLA, VC, VB, and GYA. Similarly, the drug samples were satisfactorily resolved with Rf 1.81±0.01, 0.05±0.01, 0.02±0.01, 0.09±0.01, and 0.04±0.01 for CIN, GLA, VC, VB, and GYA respectively, using HPTLC. Conclusion The importance of combining Ayurvedic formulations with contemporary high-throughput screening techniques will spark new interest in more powerful biocompatible drug leads. The findings of this study lend scientific credence to the therapeutic applications of the polyherbal formulation.

Advances in chromatography in the study of drug-plasma protein interactions

小分子药物进入人体血液循环系统后与人血清白蛋白(HSA)、α1 -酸性糖蛋白(AGP)等血浆蛋白存在广泛的相互作用,这些相互作用深刻影响药物在体内的分布及其与靶标蛋白的结合,进而影响药物效应的发挥。深入探究药物与血浆蛋白间的相互作用对于候选药物的成药性优化、新药研发、联合用药的风险评控等意义重大。而发展高效、灵敏、准确的分析检测方法是开展药物-血浆蛋白相互作用研究的关键。近年来,色谱技术由于其高通量、高分离性能、高灵敏度等特点在该领域得到了广泛的应用,包括测定血浆蛋白翻译后修饰对药物结合的影响,多种药物的竞争性结合等。其中,高效亲和色谱(HPAC)和毛细管电泳(CE)应用最为广泛,能够通过多种分析方法获取结合常数、结合位点数、解离速率常数等相互作用信息。该文着重综述了HPAC和CE在药物-血浆蛋白相互作用研究中的常用策略及最新研究进展,包括HPAC中常用的前沿色谱法、竞争洗脱法、超快亲和提取法、峰值分析法和峰衰减分析法,以及CE中常用的亲和毛细管电泳法(ACE)和毛细管电泳前沿分析法(CE-FA)等。最后,该文还对当前色谱方法存在的不足进行了总结,并对色谱技术在药物-血浆蛋白相互作用研究领域的应用前景和发展方向进行了展望。

The utility of visual appearance in predicting the composition of street opioids

Background: With the emergence of unregulated fentanyl, people who use unregulated opioids are increasingly relying on appearance in an effort to ascertain the presence of fentanyl and level of drug potency. However, the utility of visual inspection to identify drug composition in the fentanyl era has not been assessed. Methods: We assessed client expectation, appearance, and composition of street drug samples being presented for drug checking. Results of a visual screening test were compared to fentanyl immunoassay strip testing. We calculated sensitivity, specificity and likelihood ratios (LR) to assess the accuracy of the common assumption that samples with a “pebbles” appearance contain fentanyl. Results: In total, of the 2502 unregulated opioid samples tested, 1820 (73.5%) appeared as “pebbles”, of which 1729 (95.0%) tested positive for fentanyl for a sensitivity of 75.9% (95% Confidence Interval [CI]: 74.2–77.6) and specificity of 59.4% (95%CI: 57.5–61.3). Although, the odds of samples containing fentanyl was 4.60 (95%CI: 3.47−6.11) times higher among pebbles samples compared to non-pebble samples, the positive LR for pebbles to contain fentanyl was only 1.87 (CI: 1.59–2.19). The negative LR was more useful at 0.41 (95% CI: 0.36−0.46). Conclusions: A positive screening test for pebbles is not strongly enough associated to be used as a proxy for detecting fentanyl. While the absence of the appearance of pebbles does somewhat reduce the likelihood of fentanyl being present in a given sample, the high prevalence of fentanyl and fentanyl analogues in the drug supply and the risks of consumption are such that public health providers should routinely advise people who use unregulated opioids against solely relying on visual characteristics of drugs as a harm reduction strategy.

Source identification of amphetamine-like stimulants in Spanish wastewater through enantiomeric profiling

Amphetamine (AMP), methamphetamine (MAMP) and 3,4-methylenedioxymethamphetamine (MDMA) occur in wastewater not only as a result of illicit consumption, but also, in some cases, from prescription drug use or by direct drug disposal into the sewage system. Enantiomeric profiling of these chiral drugs could give more insight into the origin of their occurrence. In this manuscript, a new analytical methodology for the enantiomeric analysis of amphetamine-like substances in wastewater has been developed. The method consists of a solid-phase extraction (SPE) followed by liquid chromatography-triple quadrupole-tandem mass spectrometry (LC-MS/MS), which showed low quantification limits in the 2.4–5.5 ng L−1 range. The LC-MS/MS method was first applied to characterize a total of 38 solid street drug samples anonymously provided by consumers. The results of these analysis showed that AMP and MDMA trafficked into Spain are synthesized as racemate, while MAMP is exclusively produced as the S(+)-enantiomer. Then, the analytical method was employed to analyse urban wastewater samples collected from the wastewater treatment plants (WWTPs) of five different cities in 2018 and 2019. Consumption estimated through normalized population loads in wastewater showed an increased pattern of AMP use in the Basque Country. Furthermore, the enantiomeric profiling of wastewater samples was contrasted to lisdexamfetamine (LIS) and selegiline (SEL) prescription figures, two pharmaceuticals which metabolize to S(+)-AMP, and to R(-)-AMP and R(-)-MAMP, respectively. From this analysis, and considering uncertainties derived from metabolism and adherence to treatment, it was concluded that LIS is a relevant source of AMP in those cases with low wastewater loads, i.e. up to a maximum of 60% of AMP detected in wastewater in some samples could originate from LIS prescription, while SEL does not represent a significant source of AMP nor MAMP. Finally, removal efficiencies could be evaluated for the WWTP (serving ca. 860,000 inhabitants) with higher AMP influent concentrations. The removal of AMP was satisfactory with rates higher than 99%, whereas MDMA showed an average removal of approximately 60%, accompanied by an enrichment of R(-)-MDMA.

New psychoactive and classic substances in pooled urine samples collected at the Ultra Europe festival in Split, Croatia.

We believe that analysing pooled urine samples for recreational drugs used at mass events can provide useful information about trends in drug use. An opportunity arose with the Ultra Europe music festival, which is attended by more than 150,000 people from over 150 countries every year. We analysed 30 pooled urine samples collected from portable chemical toilets located at or close to the Ultra Europe music festival venue in Split, Croatia in 2016–2018 to detect the presence of classic and new psychoactive substances (NPS). Four urine samples collected in 2016 were from a toilet without added chemicals (otherwise used to kill the smell) while the remaining samples were collected from toilets with added chemicals. Samples were qualitatively analysed with gas chromatography-mass spectrometry (GC/MS) using the full-scan mode. Data were compared with the Wiley mass spectral library of designer drugs and our in-house library containing about 1000 compounds and metabolites. We identified forty-six different substances and metabolites, 26 of which were classic substances/metabolites, mostly from the stimulants group, while 20 were NPS. In the NPS group, most of them were phenethylamines and cathinones. The variety of substances was the highest on the first day of the festival regardless of the year, but 2018 showed a significant drop compared to the previous two years. The results of our study revealed a stable trend of classic drug consumption, while NPS trend changed from one year to another.

An electrochemical sensor for the determination of Luteolin using an alizarin red/carboxylic acid group functionalized carbon nanotube

A novel poly alizarin red (AR) film incorporating carboxylic acid group functionalized multiwall carbon nanotube (f-MWCNT) modified glassy carbon electrode (GCE) was first prepared for the determination of luteolin (Lut). Scanning and transmission electron microscopy were applied for characterization of the surface morphology of the modified electrodes and cyclic voltammetry was used to investigate the electrochemical properties of the proposed electrode towards the oxidation of Lut. Optimization of the experimental parameters was performed with regard to pH, drop-coating volume of f-MWCNT, electro-polymerization cycles of alizarin red, and different potentials of i-t. And the mechanism for Lut detection based on AR/f-MWCNT/GCE was investigated by electrochemical methods. The linearity between the oxidation peak current and the Lut concentration was obtained in the range of 0.5 to 45 μM with a detection limit of 0.17 μM (S/N = 3). In addition, the AR/f-MWCNT/GCE electrode displayed high selectivity, good stability and reproducibility, and also successfully determined Lut in drug samples.

A Simple and Sensitive LC-MS/MS Method for Determination and Quantification of Potential Genotoxic Impurities in the Vismodegib Active Pharmaceutical Ingredient

A rapid and sensitive LC-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantitative analysis of four potential genotoxic impurities Imp-A (2-chloro-5-nitroaniline), Imp-B (1-chloro-2-iodo-4-nitrobenzene), Imp-C (1-(2-chloro-5-nitrophenyl)ethan-1-one) and Imp-D (2-chloro-5-nitrobenzoic acid) in Vismodegib API drug sample. This trace analysis was achieved on CSH C18, 15.0 cm x 3.0 mm, 1.7 micron column maintained at 45°C. Optimal mobile phase consisted of 0.05% formic acid in water was used as mobile phase A and acetonitrile used as mobile phase B in gradient mode with the flow rate of 0.5 mL/min. The developed method had the short run time of 12 minutes. Quantification of four potential genotoxic impurities were carried out using mass detection with electrospray ionization in multiple reaction monitoring mode. The method was linear in the range of 0.03 ppm to 1.50 ppm for four potential genotoxic impurities with a correlation coefficient not less than 0.999. The recoveries were found satisfactory over the range between 96.67 and 106.90% for all selected impurities. The developed method was able to quantitate all four PGIs at a concentration level of 0.03 ppm (0.03 ppm with respect to 20 mg /mL Vismodegib).

Injecting Alone is More Common among Men, Frequent Injectors and Polysubstance Users in a Sample of People Who Inject Drugs

Background Injecting alone increases the risk of a fatal overdose. We examined the extent of such behavior in a sample of people who inject drugs (PWID) and the typical characteristics of those injecting alone at least once during the past four weeks. Design A cross-sectional study. PWID recruited from the street and from low-threshold services in seven Norwegian cities in September 2017 (n = 359). Associations between characteristics and injecting alone were examined using logistic regression analysis. The independent variables were gender (female/male), age, having received overdose-prevention education (no/yes), and, in the past four weeks; homelessness/shelter use (no/yes), in opioid substitution treatment (no/yes), injecting ≥ four days a week (no/yes), and substances injected (opioids only/opioids and other/other only/central stimulants (CS) only/CS and other/CS and opioids/CS, opioids and other). The adjusted odds ratios (aOR) and 95% Confidence Intervals (CI) were reported. Findings Of the 359 PWID, 84.4% reported having injected alone. Males were more likely than females to inject alone (aOR = 1.88 95% CI 1.00-3.54). Furthermore, those injecting frequently (aOR = 1.99 95% CI 1.02-3.86) and those injecting multiple substances (CS, opioids and other) (aOR = 2.94 95% CI 1.01-8.58) were more likely to inject alone compared to those injecting less frequently and opioids only. Although not statistically significant, the effect sizes in the logistic regression models suggest that polysubstance use may be driven by CS use. Conclusion Injecting alone was common in our sample of PWID, and male gender, frequent injecting and polysubstance injecting were associated with this behavior.

The efficacy of specific immunocorrection drugs against ovine oestrosis

The purpose of the research is to characterize the effect of specific immunocorrection drugs on the survival rate of the sheep botfly larvae and the immune response of the host organism. Materials and methods. Three variants of laboratory samples of specific immunocorrection drugs against sheep oestrosis have been developed based on water-soluble proteins of Oestrus ovis L. and Lucilia serricata Mg. larvae. The studies were carried out on 12 test and 3 control groups of lambs born in the current year artificially infected with 80 larvae of the sheep botfly. Fourteen days before infection, the test lambs were immunized with the prepared drugs. The drugs were injected subcutaneously in various doses (2–6 ml) and repeatedly (1–3) or by a single intranasal irrigation at a dose of 10 ml per animal. The efficacy was evaluated for the early (July – September) and summer-spring (August – April) periods of parasitism. In the test and control animals, we determined specific antibodies (JgG), stable E-rosette-forming cells (sE-RFC) and antigen-reactive (AR-RFC) T-lymphocytes. Results and discussion. The tested specific immunocorrection drugs based on somatic proteins of the sheep botfly and the sheep green bottle larvae have a pronounced protective effect against ovine oestrosis. In experiments, being affected by various drug variants, 44.6–99.1% of the sheep botfly larvae died in the early period of parasitism, and 22.6–88.1% of the sheep botfly larvae died in the summer-spring period. Subcutaneously injected drugs stimulate the produced specific antibodies and activate the effector function of the T-cell immune system to a greater extent than intranasal irrigation.

Dispersive liquid-liquid microextraction based on deep eutectic solvent for elemental impurities determination in oral and parenteral drugs by inductively coupled plasma optical emission spectrometry

A simple, fast, sensitive and green pretreatment method for determination of Cd, Co, Hg, Ni, Pb and V in oral and parenteral drug samples using inductively coupled plasma optical emission spectrometry (ICP OES) has been developed. According to United States Pharmacopoeia (USP), those metals must be reported in all pharmaceutical products for quality control evaluation (i.e., elemental impurities from classes 1 and 2A of USP Chapter 232). To improve the analytical capabilities of ICP OES, a dispersive liquid-liquid microextraction (DLLME) has performed using a safe, cheap and biodegradable deep eutectic solvent (DES) as extractant solvent (a mixture of 2:1 M ratio of DL-menthol and decanoic acid). Seven parameters affecting the microextraction efficiency have carefully optimized by multivariate analysis. Under optimized conditions, the DES-based DLLME-ICP OES procedure improved limit of quantitation (LOQ) values on range from 12 to 85-fold and afforded an enrichment factor on average 60-times higher than those obtained to direct ICP OES analysis. Consequently, LOQ values for Cd, Co, Hg, Ni, Pb and V have been on average 10-times lower than target limits recommended for drugs from parenteral route of administration. Trueness has evaluated by addition and recovery experiments following USP recommendations for three oral drug samples in liquid dosage form and three parenteral drugs. Recovery and RSD values have been within the range of 90–109% and 1–6%, respectively. All analytes were below the respectives LOQ values, hence, lower than the limits proposed by USP Chapter 232.

STABILITY INDICATING METHOD DEVELOPMENT AND VALIDATION OF MITOMYCIN AND FLUOROURACIL BY USING UPLC

Objective: The current investigation was pointed at developing and progressively validating novel, simple, responsive and stable UPLC method for the measurement of active pharmaceutical ingredients of Mitomycin and Fluorouracil. Methods: A simple, selective, validated and well-defined stability that shows isocratic UPLC methodology for the quantitative determination of Mitomycin and Fluorouracil. The chromatographic strategy utilized Inertsil ODS column of dimensions 250x4.6 mm, 5 micron, using isocratic elution with a mobile phase of acetonitrile and 0.1 percent formic acid (70:30). A flow rate of 1 ml/min and a detector wavelength of 255 nm utilizing the PDA detector was given in the instrumental settings. Validation of the proposed method was carried out according to an international conference on harmonization (ICH) guidelines. Results: LOD and LOQ for the two active ingredients were established with respect to test concentration. The calibration charts plotted were linear with a regression coefficient of R2>0.999, means the linearity was within the limit. Recovery, specificity, linearity, accuracy, robustness, ruggedness were determined as a part of method validation and the results were found to be within the acceptable range. Conclusion: The proposed method to be fast, simple, feasible and affordable in assay condition. During stability tests, it can be used for routine analysis of production samples and to verify the quality of drug samples during stability studies.

NEW STABILITY INDICATING METHOD DEVELOPMENT AND VALIDATION OF CAPECITABINE AND DOCETAXEL IN BULK AND PHARMACEUTICAL DOSAGE FORM BY USING RP-HPLC

Objective: The current investigation was pointed at developing and progressively validating novel, simple, responsive and stable RP-HPLC method for the measurement of active pharmaceutical ingredients of Capecitabine and Docetaxel. Methods: A simple, selective, validated and well-defined stability that shows gradient RP-HPLC methodology for the quantitative determination of Capecitabine and Docetaxel. The chromatographic strategy utilized Inertsil ODS column of dimensions 250x4.6 mm, 5 micron, using isocratic elution with a mobile phase of acetonitrile and water (50:50). A flow rate of 1 ml/min and a detector wavelength of 220 nm utilizing the PDA detector were given in the instrumental settings. Using the impurity-spiked solution, the chromatographic approach was streamlined. Validation of the proposed method was carried out according to an international conference on harmonization (ICH) guidelines. Results: LOD and LOQ for the two active ingredients and their impurities were established with respect to test concentration. The calibration charts plotted were linear with a regression coefficient of R2>0.999, means the linearity was within the limit. Recovery, specificity, linearity, accuracy, robustness, ruggedness were determined as a part of method validation and the results were found to be within the acceptable range. Conclusion: The proposed method to be fast, simple, feasible and affordable in assay condition. During stability tests, it can be used for routine analysis of production samples and to verify the quality of drug samples during stability studies.

Discrimination of Curcuma species from Asia using intron length polymorphism markers in genes encoding diketide-CoA synthase and curcumin synthase

Recently, Curcuma rhizome-related foods with claimed health benefits have been used worldwide; however, correct identification and quality assessment have not been conducted. Due to the wide distribution and morphological similarities of Curcuma species, the classification of some species is debated and nomenclature is inconsistent among countries. In this study, to elucidate specific molecular markers of medicinally used Curcuma species in Asia, and to solve the confusion on the reported botanical origin of crude drugs, molecular analysis based on the intron length polymorphism (ILP) in genes encoding diketide-CoA synthase and curcumin synthase and the trnK intron sequences was performed using 59 plant specimens and 42 crude drug samples from 13 Curcuma species, obtained from Asian countries. The ILP patterns of the respective species from both plant specimens and crude drug samples revealed high consistency in C. aromatica, C. zedoaria, C. phaeocaulis, C. aeruginosa, C. wenyujin, and C. zanthorrhiza, but showed intraspecies polymorphism in C. longa, C. kwangsiensis, C. amada, C. mangga and C. comosa. The C. longa specimens and samples were separated into three subgroups which were highly consistent with their geographical origins. Based on the ILP markers and the trnK intron sequences, the botanical origins of “Khamin oi” from Thailand were correctly determined to be C. longa or a hybrid between C. longa and other species, and “Wan narn kum” from Thailand and “Kasturi manjal” from India were correctly determined to be C. zanthorrhiza.

Quantitative determinations on commercial samples of Melissae folium and their antioxidant activity

Background and Aims: Melissa officinalis L. (lemon balm) is a perennial herb. Melissae folium and their preparations have been used for their sedative, spasmolytic and antibacterial actions. The study was aimed to investigate the qualities and also to compare the antioxidant activity potentials of the drug samples available in herbal markets and pharmacies in Turkey Methods: The percentages of the loss on drying and total ash were determined by gravimetric method and the percentage of total hydroxycinnamic derivatives was calculated by a spectrophotometric method according to European Pharmacopoeia. Drug samples were investigated for their potentials to scavenge the DPPH radical by using an in vitro method. Results: The percentages of the loss on drying were found to be between 8.51-16.53 %; whereas total ash amounts were determined between 9.41-11.33%. The percentage of total hydroxycinnamic derivatives was found in the range of 4.45-12.97 %. The extracts of the samples were found to have DPPH radical scavenging activity with EC50 values ranging from 10.60 to 19.10 μg/ml. Conclusion: In the assays for total ash and quantification of total hydroxy cinnamic derivatives all of the examined commercial samples were found to be compatible with standards in European Pharmacopoeia. Among the tested samples; a sample sold in pharmacy seems to have the best quality when its compared with the standards in European Pharmacopoeia.