Method For Simultaneous Analysis Research Articles

Simultaneous Determination and Comparison of Phenolic Bioactives among Three Main Kinds of Edible Chrysanthemums.

In this study, we report a simple and reliable high-performance liquid chromatography coupled with diode array detection method for simultaneous and quantitative analysis and comparison of major phenolic compounds dominant phytochemicals in Chrysanthemum morifolium, Florists chrysanthemum and snow chrysanthemum (Coreopsis tinctoria or C. tinctoria). The chromatographic separation was achieved using a reversed phase C18 column with a mobile phase of water [containing 0.1% trifluoroacetic acid (TFA)] and acetonitrile. The major phenolic compounds were completely separated within 16min at a flow rate of 1.0mL/min. Flavonoid and phenolic acid profiles of the ethanol extracts of the three flowers were analyzed. The results revealed that C. tinctoria possessed the highest amount of flavonoids (flavanomarein, flavanokanin, marein and okanin) and relative lower content of phenolic acid (chlorogenic acid and 3,5-dicafeoylquinic acid). The total content of the four flavonoids in C. tinctoria reached 53.99 ± 1.32mg/g. In particular, the marein content in C. tinctoria was as high as 36.50mg/g. Flavanomarein was only detected in C. tinctoria, whereas chlorogenic acid and 3,5-dicafeoylquinic acid were abundant in Chrysanthemum morifolium and Florists chrysanthemum. The content of marein in Chrysanthemum morifolium was slightly higher than that in Florists chrysanthemum, whereas no okanin was detected in Florists chrysanthemum under these high-performance liquid chromatography conditions. The results indicated phenolic components differ significantly depending on the cultivar, especially between C. tinctoria and common commercially available chrysanthemums. The method adopted in this study is helpful for quality control of different chrysanthemum species as well as their products, which is essential for usage and functionality clarification.

Rapid determination of acyclovir, its main metabolite 9-carboxymethoxymethylguanine, ganciclovir, and penciclovir in human serum using LC-MS/MS.

A novel MS‐based analytical method for simultaneous analysis of the antiviral drugs acyclovir, its metabolite 9‐carboxymethoxymethylguanine, ganciclovir, and penciclovir in human serum is described. These antiviral drugs are active against herpes virus infections. Acyclovir and penciclovir are regarded as safe and effective medicines with mild side effects such as headache and gastrointestinal discomfort, and ganciclovir is regarded as more toxic and is known to cause, for example, bone marrow suppression. Acyclovir’s main metabolite 9‐carboxymethoxymethylguanine is a presumptive neurotoxin and should be monitored in patients with impaired renal function or in cases with neurotoxic symptoms. A sample was prepared using protein precipitation with 1% formic acid in methanol containing isotopically labeled internal standard. Chromatographic separation on a biphenyl column and mass spectrometric detection were performed in multiple reaction monitoring (MRM) mode on a Xevo TQ‐S micro with ESI in positive ion mode, within 3 min. Inter‐day assay accuracies for the quality controls varied between 95 and 104% and intra‐day assay between 93 and 105%. Inter‐day and intra‐day assay imprecision for the quality controls ranged between 1.4 and 4.2% and 1.7 and 6.5% respectively. The lower limit of quantification for all four substances was 0.156 μmol/L. It is an accurate and reproducible method for therapeutic drug monitoring.

Quantitative Analysis of Abamectin, Albendazole, Levamisole HCl and Closantel in Q-DRENCH Oral Suspension Using a Stability-Indicating HPLC-DAD Method.

Combination therapy of many anthelmintic drugs has been used to achieve fast animal curing. Q-DRENCH is an oral suspension, containing four different active drugs against GIT worms in sheep, commonly used in Australia and New Zeeland. The anti-parasitic drugs are Albendazole (ALB), Levamisole HCl (LEV), Abamectin (ABA), and Closantel (CLO). The main purpose of this study is to present a new simultaneous stability-indicting HPLC-DAD method for the analysis of the four drugs. The recommended liquid system was 1 mL of Triethylamine/L water, adjusting the pH to 3.5 by glacial acetic acid: acetonitrile solvent (20:80, v/v). Isocratic elusion achieved the desired results of separation at a 2 mL/min flow rate using Zorbax C-18 as a stationary phase. Detection was performed at 210 nm. The linearity ranges were 15.15 to 93.75 μg/mL for ALB, 25 to 150 μg/mL for LEV, 30 to 150 μg/mL for ABA, and 11.7 to 140.63 μg/mL for CLO. Moreover, the final greenness score was 0.62 using the AGREE tool, which reflects the eco-friendly nature. Moreover, the four drugs were determined successfully in the presence of their stressful degradation products. This work presents the first chromatographic method for simultaneous analysis for Q-DRENCH oral suspension drugs in the presence of their stressful degradation products.

Development and application of a rapid and sensitive liquid chromatography-mass spectrometry method for simultaneous analysis of cytarabine, cytarabine monophosphate, cytarabine diphosphate and cytarabine triphosphate in the cytosol and nucleus

In this study, a sensitive and rapid ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the simultaneous analysis of cytarabine (ara-C), cytarabine monophosphate (ara-CMP), cytarabine diphosphate (ara-CDP) and cytarabine triphosphate (ara-CTP) in the cytosol and nucleus. The separation of analytes and endogenous interferents was achieved in 8 min on a hypercarb column (2.1 mm × 100 mm, 3 µm) by using a gradient elution with 95% acetonitrile and aqueous 5 mM hexylamine with 0.4% (v/v) diethylamine adjusted to pH 10. The analytes were detected with both negative and positive electrospray ionization in multiple reaction monitoring (MRM) mode. The calibration curve demonstrated good linearity ranging from 5 to 750 nM for ara-C, 50–7500 nM for ara-CMP, 20–3000 nM for ara-CDP and 1–150 nM for ara-CTP in the cytosol. In the nucleus, good linearity was achieved over a concentration range of 1–100 nM for ara-C, 5–500 nM for ara-CMP, 2.5–250 nM for ara-CDP and 0.5–50 nM for ara-CTP. Intra- and interbatch accuracies and precisions met the standards of validation. The matrix effect, recovery and stability were also within acceptable ranges. After incubation with 10 μM ara-C for 3 h, the levels of ara-C, ara-CMP, ara-CDP and ara-CTP in the cytosol and nucleus of HL-60 cells and HL-60/ara-C cells were determined. Most of the metabolites were found within the quantitation range. The results showed that the nuclear ara-CTP level was significantly different than the intracellular ara-CTP level between HL-60 and HL-60/ara-C cells.

Simultaneous analytical method for the carbamate pesticides in human blood or urine using LC-MS/MS for anti-food-terrorism measures

Simultaneous analytical method for the carbamate pesticides in human blood or urine using LC-MS/MS for anti-food-terrorism measures

Development and Validation of a Simultaneous Analytical Method for Six Anions in Drinking Water by Liquid Chromatography/Mass Spectrometry (LC/MS)

Development and Validation of a Simultaneous Analytical Method for Six Anions in Drinking Water by Liquid Chromatography/Mass Spectrometry (LC/MS)

Quantification and Validation of a HPLC-UV Method for Simultaneous Analysis of Nitrosoamine Impurities (NDMA, NDEA and NDIPA) in Losartan

Impurity profiling is an important aspect in drug therapy for its safety and efficacy. The study of impurities of sartans, the first line antihypertensive drugs, has become critical due to presence of cancer causing N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine (NDEA) and N-nitrosodiisopropylamine (NDIPA) in them, resulting from production and degradation process. These impurities have led to worldwide recall of products. Hence, a simple and accurate method has been developed and validated for simultaneous detection of NDMA, NDEA and NDIPA in Losartan using High Performance Liquid Chromatography - Ultra violet (HPLC - UV) system. The impurities were analyzed on Inertsil ODS 3V (250mm × 4.6mm, 5.0µm) analytical column by using water:methanol (60:40) as the mobile phase at a flow rate of 1.0 mL/min, with a run time of 30 mins. The method was developed for the acceptance limit of 0.64 ppm for NDMA, 0.177 ppm for NDEA and NDIPA respectively. On comparison with existing approaches, the developed method is fast, ideal for routine screening and is suitable for both laboratory and industrial uses. Keywords: Losartan, HPLC - UV, N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosodiisopropylamine

Fast and Environment-Friendly GC-MS Method for Eleven Organophosphorus Flame Retardants in Indoor Air, Dust, and Skin Wipes.

Organophosphorus based flame retardants (OPFRs) extensively used as alternatives to banned polybrominated diphenyl ethers and hexabromocyclododecane have been garnering interest due to the possibility that these compounds may have less significant impact on human and environmental health. Long pretreatment time, larger consumption of organic solvents, matrix interferents, and cross-contamination were found in previous studies while assessing OPFRs in indoor environments. We developed and optimized the extraction methods and simultaneous analysis of 11 OPFRs in indoor air, dust and skin wipe samples using the GC-MS approach. The proposed methods were validated using a standard addition approach, dust SRM 2585 and the real samples. Our procedures enabled the analyst to effectively limit coextracted interferences and simultaneous analytical methods of 11 target OPFRs for three matrices were achieved. The validation was performed according to standard guidelines (relative errors were identified by the analytes: −19% to 18% for indoor air, −11% to 14% for house dust, −15% to 16% for skin wipe). Good practices for quality assurance and quality control were well stated. The current high-Eco-scored methods could be categorized as “an excellent green analysis”. All analytes for the target OPFRs were detected in the real samples of indoor air, house dust and skin wipe collected from ten Taiwanese homes. Tris(2-butoxyethyl) phosphate, tris(1,3-dichloro-2-propyl)phosphate and tris(chloroisopropyl) phosphate were the most abundant OPFRs. Rapid, green and cost-effective GC-MS methods were developed and validated for the analysis of eleven OPFRs in indoor air, house dust and skin wipes.

Simultaneous Quantification of 11 Phenolic Compounds and Consistency Evaluation in Four Dendrobium Species Used as Ingredients of the Traditional Chinese Medicine Shihu.

The interchangeable use of different herbs to prepare the same formulation is a common practice in Traditional Chinese Medicine (TCM). However, this practice would require the component herbs to share similar compositions, at least in terms of the bioactive agents, to ensure they can replace each other in drug preparation. In this study, we developed an effective and comprehensive high-performance liquid chromatography-diode array detector (HPLC-DAD) method for simultaneous analysis of 11 phenolic compounds in the methanol extracts of Dendrobium huoshanense, Dendrobium nobile (D. nobile), Dendrobium chrysotoxum (D. chrysotoxum), and Dendrobium fimbriatum (D. fimbriatum), which have been identified as interchangeable ingredients for the same TCM preparation “Shihu” in the Chinese pharmacopeia (ChP). The consistency of the four Dendrobium species was evaluated on the basis of the presence of the 11 investigated compounds and the HPLC fingerprints of the methanol extracts of the plants. When gradient elution was performed with a solvent system of acetonitrile and water on a Zorbax Eclipse XDB-C18 (150 mm × 4.6 mm, 5 μm) with monitoring at 220 nm, all 11 investigated compounds were isolated at the baseline. The established HPLC method showed excellent linearity (all analytical curves showed relative coefficients [R2] > 0.999), sensitivity, precision (relative standard deviation [RSD] < 2%), and accuracy (recovery, 90.65–99.17%). These findings confirmed that the method we constructed was reliable. Quantification analysis showed significant differences in the contents of the investigated polyphenols in the four Dendrobium species. Evaluations of consistency revealed that the similarities among the four species were 0.299–0.906 in assessments based on the 11 polyphenols and 0.685–0.968 in assessments based on HPLC fingerprints. Thus, the components of the four Dendrobium species may be significantly different, and more experiments are required to determine whether they can be used interchangeably in the same amounts for preparing the formulation according to ChP.

The analysis of acidic and basic non-steroidal anti-inflammatory drugs in milk and muscle samples: a comprehensive analytical approach using UHPLC-MS/MS

ABSTRACT In animal husbandry for food production, the administration of non-steroidal anti-inflammatory drugs (NSAIDs) as anti-inflammatory, analgesic, and antipyretic agents, has seen an increase due to raised awareness of animal welfare issues. Residues of NSAIDs may be present in animal products that are intended for human consumption, and since some may pose a certain risk to human health, there is a need to monitor NSAID residues at low levels via routine and targeted surveillance. In analytical chemistry, NSAIDs are usually differentiated using their acid-base properties. Within this study, a method for simultaneous analysis of 27 NSAIDs, including both groups, in milk and muscle samples in 12.5 min is described. Sample processing consisted of enzymatic hydrolysis and acetonitrile extraction, followed by a clean-up of the extract by SPE, and measurement by UHPLC-MS/MS. The in-house validation study (alternative approach), covering trueness, precision, sensitivity, decision limit (CCα), detection capability (CCβ) and matrix effect, was designed and evaluated with the help of validation software to meet the demands of regulatory compliance. The method recovery for milk and muscle matrix was in the range of 98.1% to 106.5% and 98.8% to 102.7%, whereas the CCβ as the parameter for screening analysis, ranged from 0.07 to 46.7 µg/kg and 1.19 to 69.7 µg/kg, and the CCα, as the parameter for confirmatory analysis, from 0.11 to 56.7 µg/kg and 1.12 to 518.6 µg/kg, respectively. The occurrence of NSAID residues in milk and muscle samples was assessed using the developed method within the Croatian National Residue Control Plan, revealing quantifiable residues for diclofenac, ketoprofen, and salicylic acid mostly in milk samples. The most abundant NSAID in analysed samples was salicylic acid, which may be introduced into the food chain and be present in various types of matrices due to its natural occurrence in plants as a phytohormone.

Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines.

The antigens for acellular pertussis vaccines are made up of protein components that are purified directly from Bordetella pertussis (B. pertussis) bacterial fermentation. As such, there are additional B. pertussis toxins that must be monitored as residuals during process optimization. This paper describes a liquid chromatography mass spectrometry (LC-MS) method for simultaneous analysis of residual protein toxins adenylate cyclase toxin (ACT) and dermonecrotic toxin (DNT), as well as a small molecule glycopeptide, tracheal cytotoxin (TCT) in a Pertussis toxin vaccine antigen. A targeted LC-MS technique called multiple reaction monitoring (MRM) is used for quantitation of ACT and TCT, which have established limits in drug product formulations. However, DNT is currently monitored in an animal test, which does not have an established quantitative threshold. New approaches for DNT testing are discussed, including a novel standard based on concatenated quantitation sequences for ACT and DNT. Collectively, the method represents a “3-in-1” analytical simplification for monitoring process-related residuals during development of B. pertussis vaccines.

Usefulness of Dilution for Rapid Simultaneous Determination of Residual Pesticides in Agricultural Products by LC-MS/MS

A rapid, easy and versatile, simultaneous analytical method by LC-MS/MS based on extraction of QuEChERS method (EN 15662:2008) was developed for the determination of residual pesticides in agricultural products. In this method, it allowed to prepare a test solution only dilution without purification using solid phase extraction column. The method was assessed for 210 pesticide residues in 21 kinds of vegetables and fruits at the fortification levels of 0.01 and 0.1 μg/g according to the method validation guideline (Ministry of Health, Labour and Welfare of Japan). As a result, 194 to 210 analytes met the management criteria of the guideline. Thus, the present method could be useful for a rapid simultaneous determination of residual pesticides in agricultural products.

Simultaneous analysis of urinary total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol, N\u2032-nitrosonornicotine, and cotinine by liquid chromatography-tandem mass-spectrometry

Biomarkers of exposure to harmful tobacco constituents are key tools for identifying individuals at risk and developing interventions and tobacco control measures. However, tobacco biomarker studies are scarce in many parts of the world with high prevalence of tobacco use. Our goal was to establish a robust method for simultaneous analysis of urinary total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), N′-nitrosonornicotine (NNN), and cotinine at the Advanced Centre for Treatment, Research and Education in Cancer (ACTREC) in Mumbai, India. These biomarkers are validated measures of exposure to the carcinogenic tobacco nitrosamines 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and NNN and the addictive alkaloid nicotine, respectively. The established method is characterized by excellent accuracy, linearity, and precision, and was successfully applied to the analysis of 15 smokeless tobacco (SLT) users and 15 non-users of tobacco recruited in Mumbai. This is the first report of establishment of such procedure in a laboratory in India, which offers the first in-country capacity for research on tobacco carcinogenesis in Indian SLT users.

Analysis of furans and pyridines from new generation heated tobacco product in Japan

BackgroundIn recent years, heated tobacco products (HTPs), which are widely used in Japan, have been sold by various brands using additives such as flavors. It has been reported that the components of mainstream smoke are different from those of conventional cigarettes. In this study, we established an analytical method for furans and pyridines in the mainstream smoke, which are characteristic of HTPs and particularly harmful among the generated components, and investigated the amount of component to which the smokers are exposed.MethodsWe established a simple analytical method for simultaneous analysis of gaseous and particulate compounds in the mainstream smoke of HTPs (IQOS, glo, ploom S) in Japan by combining a sorbent cartridge and glass fiber filter (Cambridge filter pad (CFP)). Both the sorbent cartridge and CFP were extracted using 2-propanol and analyzed via GC-MS/MS to determine the concentration of furans and pyridines generated from each HTP.ResultsThe results showed that the levels of target furans such as furfural, 2-furanmethanol, 2(5H)-furanone, and 5-methylfurfural tended to be higher in the mainstream smoke of glo than in standard cigarettes (3R4F). Pyridine, which is generated at a high level in 3R4F as a combustion component, and 4-ethenylpyridine (EP), which is a known marker of environmental tobacco smoke, were detected. Among these components, 2-furanmethanol and pyridine are classified as Group 2B (possibly carcinogenic to humans) by the International Agency for Research on Cancer (IARC). Therefore, it is possible that they will contribute to the health effects caused by use of HTPs.ConclusionsUsing the new collection and analytical method for furans and pyridines in the mainstream smoke of HTPs, the level of each compound to which smokers are exposed could be clarified. By comprehensively combining information on the amount of ingredients and toxicity, it will be possible to perform a more detailed calculation of the health risks of using HTPs. In addition, the components detected in this study may be the causative substances of indoor pollution through exhaled smoke and sidestream smoke; therefore, environmental research on the chemicals generated from HTPs would be warranted in future studies.

Development of a Novel and Stability Indicating RP-HPLC-UV Method for Simultaneous Analysis of Carbetocin and Ten Impurities in Carbetocin Injection Products

Development of a Novel and Stability Indicating RP-HPLC-UV Method for Simultaneous Analysis of Carbetocin and Ten Impurities in Carbetocin Injection Products

Usefulness of Dilution for Rapid Simultaneous Determination of Residual Pesticides in Edamame (Immature Soybeans) by LC-MS/MS

A rapid, easy and versatile, simultaneous analytical method by LC-MS/MS based on extraction of QuEChERS method (EN 15662:2008) was developed for the determination of residual pesticides in Edamame (immature soybeans). In this method, it allowed to prepare a test solution only dilution without purification using solid column. As a result, 202 analytes met the management criteria of the guideline for validity assessment (Ministry of Health, Labour and Welfare of Japan). Thus, the present method could be useful for a rapid simultaneous determination of residual pesticides in green soybean.

Comprehensive profile of DNA adducts as both tissue and urinary biomarkers of exposure to acrylamide and chemo-preventive effect of catechins in rats

Two representative DNA adducts from acrylamide exposure, N7-(2-carbamoyl-2-hydroxyethyl) guanine (N7-GA-Gua) and N3-(2-carbamoyl-2-hydroxyethyl) adenine (N3-GA-Ade), are important long-term exposure biomarkers for evaluating genotoxicity of acrylamide. Catechins as natural antioxidants present in tea possess multiple health benefits, and may also have the potential to protect against acrylamide-induced DNA damage. The current study developed an ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC−MS/MS) method for simultaneous analysis of N7-GA-Gua and N3-GA-Ade in tissues and urine. The validated UHPLC−MS/MS method showed high sensitivity, with limit of detection and limit of quantification ranging 0.2–0.8 and 0.5–1.5 ng/mL, respectively, and achieved qualified precision (RSD<14.0%) and spiking recovery (87.2%–110.0%) with elution within 6 min, which was suitable for the analysis of the two DNA adducts in different matrices. The levels of N7-GA-Gua and N3-GA-Ade ranged 0.9–11.9 and 0.6–3.5 μg/g creatinine in human urine samples, respectively. To investigate the interventional effects of catechins on the two DNA adducts from acrylamide exposure, rats were supplemented with three types of catechins (tea polyphenols, epigallocatechin gallate, and epicatechin) 30 min before administration with acrylamide. Our results showed that catechins effectively inhibited the formation of DNA adducts from acrylamide exposure in both urine and tissues of rats. Among three catechins, epicatechin performed the best inhibitory effect. The current study provided evidence for the chemo-preventive effect of catechins, indicating that dietary supplement of catechins may contribute to health protection against exposure to acrylamide.

Validated Stability-Indicating Assay UHPLC Method for Simultaneous Analysis of Saxagliptin and Metformin in Fixed-Dose Combinations

A Saxagliptin (SAX) and Metformin (MET) fixed-dose combination (FDC) can improve the chances of achieving optimal glycaemic control in type 2 diabetes mellitus subjects. Accordingly, the scientific novelty of design work was to develop a specific and precise stability-indicating UHPLC assay method for simultaneous quantification of cited drugs in extended-release FDC's. The reversed-phase UHPLC resolution was analyzed with the assistance of UPLC BEH C18 (150 mm × 2.1 mm) with 1.7 µm particle size column at ambient temperature using a solvent system in a proportion of (85:15 % v/v) potassium dihydrogen orthophosphate buffer and acetonitrile, with 0.4 mL/min rate of flow of a solvent system. The analytes were supervised at 220 nm by employing photodiode array recognition. The retention times of analytes SAX and MET were 2.687 ± 0.022 min and 1.856 ± 0.03 min, respectively. The SAX and MET have confirmed the linearity ranges of 10 – 60 μg/mL and 100 – 600 μg/mL, with 0.9989 and 0.9979 determination coefficients. The UHPLC method was effectually validated concerning the accuracy, precision, sensitivity, robustness, ruggedness, selectivity, and specificity. Moreover, the anticipated UHPLC method's capability to analyzed the SAX and MET with no obstruction from degradation products.

Validation Method for Simultaneous Analysis of Betamethasone Dipropionate and Retinoic Acid in Cream Formulation by Ultra High-Performance Liquid Chromatography

A simple, precise and rapid reverse phase UHPLC-PDA method has been developed and validated for the simultaneous analysis of betamethasone dipropionate (BTM) and retinoic acid (REA) in whitening cream dosage form. The mixture of betamethasone dipropionate (BTM) and retinoic acid (REA) was separated on Phenomenex Bonclone 10 C18 (150 mm x 3.9 mm) column. All separations were performed with a Dionex 3000 Photodiode Array (PDA) detector on 240 nm and 340 nm wavelength, column temperature at 40oC, and flow rate at 1.0 ml/min. The mobile phase was methanol-acetic acid 0.1% (85:15) with pH 5. The retention times of BTM and REA were found to be 1.90 and 3.70 minutes, respectively. The method showed good linearity with correlation coefficients of 0.9998 and 1.0000 over the ranges of 5-25 ppm for BTM and 25-200 ppm for REA, respectively. The method has mean recoveries in the range of 99.37% to 100.94% for all analytes. The developed method can be used in the routine analysis of BTM and REA in cream formulation, as well as for qualitative analysis in whitening cosmetic preparations or for quantitative analysis in drug preparations.

Front Cover: Capillary electrophoresis method for simultaneous analysis of caffeine, vanillin and ethyl vanillin in beverages

Front Cover: Capillary electrophoresis method for simultaneous analysis of caffeine, vanillin and ethyl vanillin in beverages