Method Validation Characteristics Research Articles

Chemometric assisted natural DES based VA-DLLME-LC-MS/MS method for the quantitative determination of Garcinol in biofluids/tissues: A practical application to pharmacokinetics and biodistribution studies

Garcinol (GAR) is a polyisoprenylated benzophenone obtained from Garcinia indica used as anti-oxidant and anti-inflammatory in traditional medicine and due to these activities, it possesses anticancer properties. It is considered to be a next generation epigenetic drug. A green solvent based analytical method which is efficient, sophisticated, and highly enriched has been developed for the quantitative analysis of GAR in biological samples (plasma, liver, kidney and spleen) with the use of deep eutectic solvent (DES) for its extraction. A series of 23 DESs were synthesized and out of which, Thymol (Th)-Terpeniol (T), 2:1 molar ratio with a more hydrophobic environment and high interaction efficiency between GAR and DES was identified for the better extraction from mice plasma and tissue samples. The Design of Experiment approaches like placket-burmann design and central composite design were used to optimize the method conditions. The method validation characteristics, such as limit of detection (0.193–0.237 ng/mL), limit of quantification (0.644–0.697 ng/mL), lower limit of quantification (0.5 ng/mL), broad range of linearity with R2 (0.9994–0.9997) with a percent recovery not less than 87% was observed, which are well within the acceptance criteria for a bioanalytical method. The enrichment factor is upto 53–60 folds, with high extraction efficiency (89–97%). The measurement uncertainty was estimated with an expanded uncertainty ranged between 10.9%–19.0%. The method developed and validated was effectively applied to examine the pharmacokinetic and biodistribution patterns for GAR in mice.

Method Validation Approaches for Analysis of Constituents in ENDS.

We assessed how many peer-reviewed publications reporting chemical quantities and/or yields from electronic nicotine delivery systems (ENDS) have included adequate method validation characteristics in the publication for appropriate interpretation of data quality for informing tobacco regulatory science. We searched 5 databases (Web of Knowledge, PubMed, SciFinder, Embase, EBSCOhost) for ENDS publications between January 2007 and September 2018. Of the 283 publications screened, 173 publications were relevant for analysis. We identified the publications that report a certain degree of control in data quality, ie, the publications that report marginally validated methods (MVMs). MVMs refer to the methods that: (1) report 3 or more International Conference on Harmonisation (ICH) method validation characteristics, (2) state the method was validated, (3) cite their own previous publication(s) that report MVMs, or (4) use a method within the accreditation scope of an accredited laboratory. Overall, 97 publications (56%) report MVMs in their studies. This percentage also reflects the publication distribution for the majority of the 28 chemicals measured by MVMs. This study highlights the need for reporting sufficient validation characteristics following appropriate guidance to ensure the accuracy and reliability of the published analytical data for proper data interpretations that may support policy.

Development and Validation of UV-Spectrophotometric method for the Estimation of Curcumin in Standardised Polyherbal Formulations

Objective: We aimed to develop and validate a simple UV-spectrophotometric method relates to the estimation of curcumin in standardised polyherbal formulations. Methods: Pure curcumin was received as generous gift sample from Andhra University, India, all solvents and reagents of analytical reagent (AR) grade were procured from local suppliers and readily used for the method development. The method validation parameters were evaluated as per International Conference on Harmonization (ICH) guidelines. Further, this method was applied for the assay of curcumin in marketed product Live-well™, CUMIN (standardised turmeric extract (STE) fortified with standardised black pepper (SBE) extract) capsules using UV-spectrophotometric method. Results and Discussion: It was confirmed from the results of method validation characteristics such as specificity, linearity, range, precision, accuracy, and robustness were keeping within the limits of acceptance criteria defined under ICH guidelines, 76.0560% of curcumin was estimated to be present in each Live-well™, CUMIN capsule containing a minimum 95% of the total curcuminoids (label claimed). Conclusion: In summary, our method was found to be reliable in detecting the unknown percentage of curcumin in Live-well™, CUMIN capsule formulation. Also, the method was proved to be specific, linear, precise, accurate and robust to use for routine analysis of polyherbal formulations containing curcumin as their principal component in the extracts. Key words: UV-spectrophotometric method, Curcumin, ICH Guidelines, Live-well™, CUMIN.

РОЗРОБКА ТА ВАЛІДАЦІЯ СПЕКТРОФОТОМЕТРИЧНОЇ МЕТОДИКИ КІЛЬКІСНОГО ВИЗНАЧЕННЯ ЕНІСАМІУМУ ЙОДИДУ В КАПСУЛАХ

Мета роботи. Розробити методику кількісного визначення енісаміуму йодиду в капсулах із застосуванням методу абсорбційної спектрофотометрії в ультрафіолетовій області.Матеріали і методи. Використано стандартний зразок енісаміум йодиду та зразок готової лікарської форми виробництва ПАТ «Фармак». У ході роботи було встановлено оптимальну робочу концентрацію енісаміуму йодиду 0,02 мг/мл. Дослідження проводили із застосуванням спектрофотометра «Specord 250 Plus».Результати й обговорення. Продемонстровано, що оптимальна довжина хвилі при визначенні становить 267 нм. Вивчено валідаційні характеристики методики в діапазоні 0,016-0,024 мг/мл (80-120%): специфічність, правильність, прецизійність, лінійність, діапазон застосування, робасність, внутрішньолабораторна прецизійність відповідно до рекомендацій Міжнародної конференції з гармонізації. Коефіцієнт кореляції (r) між введеним та знайденим значенням для енісаміуму йодиду становить 0,99996, а відносний довірчий інтервал (Dz) 0,23 %. Підтверджено стабільність розчинів при їх зберіганні при кімнатній температурі протягом 48 годин.Висновки. Розроблено спектрофотометричну методику кількісного визначення енісаміуму йодиду в капсулах, експериментально доведено, що вона придатна для контролю якості капсул енісаміуму йодиду.

ВАЛІДАЦІЯ МЕТОДИКИ КІЛЬКІСНОГО ВИЗНАЧЕННЯ КРОМОГЛІКАТУ НАТРІЮ В ОЧНИХ КРАПЛЯХ ПРОТИАЛЕРГІЙНОЇ ДІЇ

<p align="left"><strong>Validation</strong><strong> </strong><strong>studies</strong><strong> </strong><strong>of</strong><strong> </strong><strong>cromoglicate</strong><strong> </strong><strong>sodium</strong><strong> </strong><strong>assay</strong><strong> </strong><strong>at</strong><strong> </strong><strong>eye</strong><strong> </strong><strong>drops</strong><strong> </strong><strong>with antiallergic action</strong><strong></strong></p><p><strong>Е. G. F</strong><strong>etisova</strong></p><p>NationalUniversityofPharmacy,Kharkov</p><p><strong>Introduction.</strong> Today during pharmaceutical development of drugs as well as at the all stage of product lifecycle, significant sense obtain analytical supporting of experiments and estimation of analytical methods which are necessary for control of process and finished products. The analytical methods, conditions of their carrying out, reagents and standard samples should be standardized with aim to obtain reproducing results that are required at the carrying out of drug pharmaceutical development and at the composition of drug registration materials. In accordance with the requirements of the State Pharmacopoeia of Ukraine (SPU) all methods of drug quality control should be validated and proved experimentally that the method is available for intended purposes. We have developed eye drops with antiallergic action based on cromoglicate sodium (CS) and the method of quantitative determination of cromoglicate sodium at the eye drops by the spectrophotometric method. The aim of the work is the determination of the method validation characteristics and experimental evidence that the method can make reliable and reproducing results for the test “Assay”.</p><p><strong>Materials and methods</strong>. The drug in the form of eye drops based on CS in the therapeutic concentration of 2% was the object of study. CS produced by company "Fermion" (Finland) and conformed to European Pharmacopoeia, was used for preparing of the eye drops. Absorption spectrophotometry in the UV region (SPU, 2.2.25) was the method of study. Analytical equipment: spectrophotometer UV-VIS HP 8453, company "Hewlett Packard" (USA); laboratory digital scales VA-210S, firm "Sartorius" (Germany); measuring vessels accuracy class A.</p><p><strong>R</strong><strong>esults and discussion</strong>. Validation of the developed method of quantitative determination of CS have been conducted in accordance with the requirements of SPU by major validation characteristics: specificity, accuracy, precision (convergence), linearity, range, intermediate precision. Also, the calculation of estimated total uncertainty of the analysis have been carrying out. According to the requirements of SPU the concentrations range for the study of linearity, accuracy and convergence has been ranged from 80 % to 120 % in increments of 5 %, the eligibility criteria was designed for B = 5 %, so the maximum uncertainty analysis should not exceed 1.6 %. Method characterized by sufficient convergence and accuracy throughout the range of concentrations studied. Found the relative size of the confidence interval (Δ<sub>Ζ</sub> = 1,23 %) is less than the critical value for the convergence of the results (1,6 %). Systematic error of the method (δ = 0,04 % < 0,41) is almost insignificant. Specificity is confirmed by insignifican deposit "placebo" in the total amount of background absorption compared with the maximum allowable uncertainty analysis (∆<sub>As</sub> = 1,6 %) and excipients do not interfere with the quantitative determination of CS in the preparation. Intermediate precision of the analysis have been confirmed by the compliance of the relative confidence interval for 6 replicates of a series of eye drops with 2 % CS (Δ<sub>Ζ</sub> = 0,62 %) to acceptance criteria (≤ 1,6 %). Full predicted uncertainty of the test results for the "Assay" of cromoglicate sodium (Δ<sub>As </sub>= 1,42 %) is less than critical value (1,6 %), so the method will give correct results in other laboratories on this indicator.</p><p><strong>Conclusion</strong>. The method of quantitative determination of CS at the eye drops by the spectrophotometric method has been validated in accordance with the requirements of the State Pharmacopoeia of Ukraine. For the test it has been established accordance to specified acceptance criteria for the assay limits ±5% by the validation parameters: specificity, linearity, precision (precision), accuracy within the using range (80-120% of the nominal contents). The complete uncertainty forecast of the method was carried out in order to confirm it correctness when reproduce at other laboratories and it was established that the projected total uncertainty of the results wasn’t exceed the required acceptance criteria. Based on the results of validation studies it has been substantiated and verified experimentally that the test can make reliable results, can be correctly reproduced and be suitable for quality control of 2% eye drops based on CS during differing stage of product lifecycle.</p>

Development and Validation of Stability Indicating LC-PDA Method for Mycophenolate Mofetil in Presence of Mycophenolic Acid and Its Application for Degradation Kinetics and pH Profile Study

Factorial design tool applied for development of isocratic reversed-phase stability-indicating HPLC method for the analysis of mycophenolate mofetil (MMF) and its degradation products. MMF stress degradation products mycophenolate acid (MPA) and DP3 (USP impurity H) were isolated and used for quantitation. Separation achieved on a Symmetry C18 (250 mm × 4.6 mm, 5.0 μ) column using a methanol: acetate buffer (75 : 25 v/v), pH 6.0 (adjusted with acetic acid), at 0.5 mL flow rate, column maintained at 55°C, and data integrated at 251 nm. MMF is subjected to hydrolysis, oxidation, heat degradation, and so forth; under all these conditions degraded products are well separated. The method validation characteristics included accuracy, precision, linearity, range, specificity, and sensitivity. Robustness testing is conducted to evaluate the effect of minor changes to the chromatographic conditions and to establish appropriate system suitability parameters. The proposed method is used to investigate kinetics of acid, alkali hydrolysis and oxidation process. Major degradation products MPA and DP3 were isolated and quantitated. Characterization of MPA by NMR and LC-MS/MS and other degraded products by LC-MS/MS is attempted successfully. The method is used successfully for the quality assessment of three MMF drug commercial formations and its acid, alkali, and oxidative degradation kinetics study.

Development and Validation of a Stability-Indicating HPLC-Photodiode Array Detector Method for Formulation Analysis and Degradation Kinetics and Dissolution Studies of Mycophenolate Sodium and HPLC/MS/MS Characterization of its Stress Degradation Products

Abstract A simple stability-indicating isocratic RP-HPLC method was developed and validated for the determination of mycophenolate sodium and its alkali degradation product. Forced degradation of the drug was carried out under thermolytic, photolytic, acid/base hydrolytic, and oxidative stress conditions. Alkali degradation product DP1 was isolated, and separation of stress degradation products was achieved on a Symmetry C18 (250 × 4.6 mm × 5.0 μm) column using the mobile phase methanol–acetate buffer adjusted with acetic acid to pH 6.0 (76 + 24, v/v) at a 0.55 mL/min flow rate and 50°C. Data were integrated at the detection wavelength of 251 nm. The method validation characteristics included accuracy, precision, linearity, range, specificity, and sensitivity per International Conference on Harmonization guidelines. Robustness testing was conducted to evaluate the effect of minor changes in the chromatographic conditions and to establish appropriate system suitability parameters. Structural elucidation of degraded products was performed by HPLC/MS/MS. The method was used successfully for drug product analysis, dissolution study, and determination of the drug's acid, alkali, and oxidative degradation kinetics.

The impact of decreased bead count to determine concentrations of amyloid beta1‐42, total‐tau, and phosphorylated‐tau181 in human cerebrospinal fluid using xMAP technology

Alzheimer's disease is the leading cause of human dementia. The lack of diagnostic tests and limited therapeutic options has driven the search for endogenous biomarkers. The INNO‐BIA AlzBio3 assay is a multiplex flow‐based immunoassay measuring Aβ42, tau, and p‐tau in cerebrospinal fluid (CSF). This study assesses assays performance under varying bead count (BC) parameters. Original method validation parameters at 100 BC were acceptable. Reanalyses performed at 3, 10, 25, and 50 BCs were compared to 100 BC data by ANOVA, Bland–Altman analysis, evaluation of concordance correlation coefficients, and frequency distribution of coefficient of variation (CV) ranges. Method validation characteristics were acceptable with 100 BCs. Equivalency for 25 and 50 versus 100 BCs was demonstrated, but not for 3 and 10 BCs. A general trend of decreasing agreement between decreasing BCs and the 100 BC reference resulted in decreases in concordance coefficients ρc. The frequency of CV values greater than 20% increased with decreasing BCs, and CV values of 5% or less decreased with decreased BCs. Statistical analyses demonstrate that BCs of 3 and 10 are not equivalent with the reference and should not be used as a basis for determination of Aβ42, tau, and p‐tau concentration in human CSF. © 2011 Wiley‐Liss, Inc. and the American Pharmacists Association J Pharm Sci 100:4655–4663, 2011

Method validation and qualification of instruments for atomic spectrometry

This work describes the analytical procedures for atomic absorption and inductively coupled plasma (ICP) techniques that have to be used in order to obtain a license to sell drugs in the USA. The qualification of atomic absorption spectrometers and ICP instruments is described. The method validation characteristics, e.g., accuracy, precision, linearity, range, detection limits, and quantification are discussed. The time involved and the quality of documentation are pointed out. The consequences for laboratory personnel and operating costs are also discussed.