Development and validation of UPLC-MS/MS method for 5-Fluorouracil quantification in dried blood spot.

5-Fluorouracil is an antimetabolite drug indicated for cancer treatment. Therapeutic drug monitoring of 5-Fluorouracil is necessary because 5-Fluorouracil has narrow therapeutic window and its concentration in blood is affected by individual conditions, like gene polymorphisms. Dried Blood Spot (DBS) is one of the biosampling methods used for therapeutic drug monitoring. Asides from reducing patients' discomfort, the use of DBS can increase 5-Fluorouracil stability by stopping the enzymes activity in blood. Therefore, this research developed a method to monitor 5-Fluorouracil levels in DBS using ultra-high performance liquid chromatography-tandem mass spectrometry. Sample preparation was carried out by extracting DBS using 2-Propanol: ethyl acetate (16:84). Reconstituted samples were analyzed using ultra high performance liquid chromatography equipped with Acquity® UPLC BEH C18 column (2.1 × 100 mm; 1.7 µm). The ionization process was carried out in negative electrospray ionization mode. Multiple Reaction Monitoring (MRM) values were set at m/z 128.97 > 41.82 for 5-Fluorouracil and 168.97 > 57.88 for propylthiouracil as the internal standard. Optimum analytical conditions were obtained with acetonitrile-ammonium acetate 1 mM (95:5) as mobile phase, flow rate of 0.15 mL/min, and column temperature of 40 °C. The lowest level of quantification obtained from this method was 0.1 µg/mL with a calibration curve range of 0.1 µg/mL-60 µg/mL. This method was proven to be valid according to the requirements set by the US Food and Drug Administration and the European Medicines Agency.

Ultrasonic-Assisted Extraction of Xanthorrhizol from Curcuma xanthorrhiza Roxb. Rhizomes by Natural Deep Eutectic Solvents: Optimization, Antioxidant Activity, and Toxicity Profiles.

Xanthorrhizol, an important marker of Curcuma xanthorrhiza, has been recognized for its different pharmacological activities. A green strategy for selective xanthorrhizol extraction is required. Herein, natural deep eutectic solvents (NADESs) based on glucose and organic acids (lactic acid, malic acid, and citric acid) were screened for the extraction of xanthorrhizol from Curcuma xanthorrhiza. Ultrasound-assisted extraction using glucose/lactic acid (1:3) (GluLA) gave the best yield of xanthorrhizol. The response surface methodology with a Box-Behnken Design was used to optimize the interacting variables of water content, solid-to-liquid (S/L) ratio, and extraction to optimize the extraction. The optimum conditions of 30% water content in GluLA, 1/15 g/mL (S/L), and a 20 min extraction time yielded selective xanthorrhizol extraction (17.62 mg/g) over curcuminoids (6.64 mg/g). This study indicates the protective effect of GluLA and GluLA extracts against oxidation-induced DNA damage, which was comparable with those obtained for ethanol extract. In addition, the stability of the xanthorrhizol extract over 90 days was revealed when stored at -20 and 4 °C. The FTIR and NMR spectra confirmed the hydrogen bond formation in GluLA. Our study reported, for the first time, the feasibility of using glucose/lactic acid (1:3, 30% water v/v) for the sustainable extraction of xanthorrhizol.

Development and validation of ivermectin quantification method in volumetric absorptive microsampling using liquid chromatography-tandem mass spectrometry.

Background: Ivermectin is a broad-spectrum anthelmintic used to control onchocerciasis from nematode parasites. As an anthelmintic, ivermectin is designed to have high levels in the gastrointestinal tract, so that the systemic intake is relatively low. Due to the very small concentration of ivermectin, a selective and sensitive approach is needed for the analysis of ivermectin in blood. Several methods have been developed using plasma and Dried Blood Spots, but there are still shortcomings due to hematocrit effects. Therefore, this study was conducted to establish a validated ivermectin analysis method with doramectin as the internal standard in using Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry. Methods: Mass spectrometry equipped with triple quadrupole and positive electrospray ionization mode was used to conduct the analysis. For the biological matrix, whole blood was used by Volumetric Absorptive Microsampling and extracted using a protein precipitation technique with a combination of acetonitrile and methanol (1:1). VAMS has some advantages such as not being affected by hematocrit, requires a small and fixed volume of sample, also a more efficient sampling process. Results: The optimum conditions were achieved with an Acquity® UPLC BEH C18 column (1,7 µm; 2.1 × 100 mm); extracted-flow rate was 0,2 mL/min; mobile phase was 5 mM ammonium formate pH 3.00 and acetonitrile (10:90) with isocratic elution. Multiple Reaction Monitoring (MRM) detection by m/z values was 892.41 > 569.5 for ivermectin and 916,41 > 331,35 for doramectin. Conclusion: The method has been appropriately validated in compliance with the 2018 guidelines laid out by the US Food and Drug Administration. Resulting the minimum detection (LLOQ) was 1 ng/mL with a linear concentration range spanning from 1 to 150 ng/mL.

Validation of bioanalytical method for quantification of Vitamin K2 (MK-4) in human plasma by high-performance liquid chromatography-ultraviolet.

Vitamin K can reduce warfarin's anticoagulant action, causing a variance in response among individuals taking warfarin. Vitamin K comes in two forms, namely Vitamin K1 (phylloquinone) and K2 (menaquinones). Menaquinone-4 (MK-4) is a kind of Vitamin K2 found in meat and dairy products. Analysis of MK-4 levels in human plasma is very useful for patients who receive warfarin therapy. High-performance liquid chromatography (HPLC) can be used for warfarin's bioanalysis, and it must be validated. The purpose of this study was to validate the bioanalytical method for quantification of Vitamin K2 (MK-4) in human plasma according to the 2019 European Medicines Agency (EMA) guideline. Vitamin K2 (MK-4) was extracted using acetonitrile. HPLC with an ultraviolet detector at 245 nm, using a T3 column set at 30°C and an isocratic mobile phase containing methanol: phosphate buffer (95:5) at pH 3, a flow rate of 1 mL/min was used in this study. The warfarin concentration of 0.5-3 µg/mL was used. About 5.50%-17.42% and 6.18%-8.74%, respectively, were the average ranges of percentage coefficient of variation and percentage difference. There was no response at the analyte's retention time in the six blank plasmas and at the analyte's retention time in the blank after the injection of upper limit of quantification, indicates that the procedure was very selective and did not result in any carryover. This bioanalytical method fulfills the parameters of selectivity, accuracy, precision, and carryover based on the 2019 EMA guidelines.

Development of a Colorimetric Paper Sensor for Hg2+ Detection in Water Using Cyanuric Acid-Conjugated Gold Nanoparticles.

Hg2+ is one of the most dangerous pollutants that can cause damage to organs and the immune system. The common detection methods of Hg2+ require sophisticated instrumentation and a long time for analysis. The purpose of this study was to develop a sensor for the detection of Hg2+ using filter paper immobilized by gold nanoparticles (AuNPs) conjugated with cyanuric acid (CA). The clear color change from pink to bluish purple is the response of the CA-AuNPs filter paper sensor to exposure to Hg2+. Detection can be observed visually with the naked eye and/or with imageJ software; the detection limit is 0.05 µM. The colorimetric response of the sensor was also selective towards Hg2+ after testing with different metal ions. In addition, the response from the sensor was also consistent for lake water samples spiked with Hg2+. The results of this research provide a promising basic technology for the development of sensors that are affordable, fast, portable, and easy to use for the detection and monitoring of Hg2+ levels in water.

Validation of warfarin enantiomer analysis method in plasma using high-performance liquid chromatography fluorescence detector.

Warfarin (WF) is an anticoagulant commonly used for thromboembolism-related diseases. This study aims to assess the pharmacokinetic profile of WF. The stereospecific interaction of S-and R-WF requires quantification of the enantiomer to determine the pharmacokinetic profile. The analysis method of the enantiomers in plasma is developed using an HPLC fluorescence detector with a Chiralcel OD-RH column (4.6 mm × 150 mm i.d., 5 m) and a Chiralcel OD-RH guard column (4.0 mm × 10 mm, 5 m). The separation is conducted using isocratic with acetonitrile mobile phase: Phosphate buffer, pH 2.00 (40:60 v/v), column temperature 40°C, flow rate 1 mL/min, injection volume 50 L. WF is measured at an excitation wavelength of 310 nm and emission of 350 nm. This method results in limit of detection (LOD) values of 18.6 ng/mL and limit of quantitation (LOQ) of 62.01 ng/mL for R-WF and LOD values of 18.61 ng/mL and LOQ of 62.04 ng/mL for S-WF. The results showed a linearity in concentration between 100 and 2500 ng/mL with r 2 = 0.9969 and r 2 = 0.9991 for R-and S-WF. The validation requirements of selectivity, accuracy, and precision for within and between run with a value of <15% for % relative standard deviation and % diff were achieved. This method can be used in the sample measurement of WF pharmacokinetic studies.

A Review of Analytical Methods for Codeine Determination.

Codeine is derived from morphine, an opioid analgesic, and has weaker analgesic and sedative effects than the parent molecule. This weak opioid is commonly used in combination with other drugs for over-the-counter cough relief medication. Due to the psychoactive properties of opioid drugs, the easily obtained codeine often becomes subject to misuse. Codeine misuse has emerged as a concerning public health issue due to its associated adverse effects such as headache, nausea, vomiting, and hemorrhage. Thus, it is very important to develop reliable analytical techniques to detect codeine for both quality control of pharmaceutical formulations and identifying drug misuse in the community. This review aims to provide critical outlooks on analytical methods applicable to the determination of codeine.

Correlation study between gastronomy culture and content of selenium towards prevalence of cardiovascular and diabetes in West Java, Indonesia.

BACKGROUND AND AIM: Selenium (Se) is an important element in the human body. Deficiency or excess of Se can cause harm to human health. A previous study showed an association of Se with cardiovascular and diabetes diseases. One of the food sources of Se is vegetables. In West Java, Indonesia, people consume fresh vegetables such as Garlic, Jengkol, and Petai. This research aims to study the correlation between the gastronomy culture of people in West Java, Se content in Garlic (Allium sativum), Jengkol (Archidendron pauciflorum) and Petai (Parkia speciosa) from several Regencys/cities in West Java, and the prevalence cardiovascular and diabetic diseases. METHOD: A cultural study was conducted based on a literature review. Cluster sampling was chosen for the sampling method. The prevalence of cardiovascular disease and diabetes in these regencies were obtained from the Ministry of Health of Indonesia. The measurement of Se content in a sample was conducted by the fluorometry method, based on the formation of the piazoselenol complex from the reaction between selenite ion and DAN (2,3-diaminonapthalene). RESULTS: People in West Java prefer to consume garlic, jengkol, and petai as a fresh vegetable as part of their culture. The highest content of Se in Allium sativum was found in Tasikmalaya City with a value of 69.20 ng/g. For Archidendron pauciflorum from Subang Regency values were 498 ng/g. Parkia speciosa found in the Bandung Barat Regency had a mean value 257.9 ng/g. There is a positive correlation between Se-concentration in Archidendron pauciflorum and the prevalence of diabetes while negative correlation with the prevalence of cardiovascular disease. In addition, no correlation was observed for Allium sativum and Parkia Specose might be due to a lower Se-concentration in these vegetables that in the Archidendron fauciflorum. CONCLUSION: Different areas have varying concentrations of Se in plants that grow in the region. The gastronomy culture and Se content may play a role to increase or decrease cardiovascular and diabetes prevalence in that area.

Extraction of atenolol from spiked blood serum using a molecularly imprinted polymer sorbent obtained by precipitation polymerization.

Atenolol (ATE) is a cardio-selective ß-blocker that is used in the treatment of hypertension over extended periods. However, ATE, like propranolol, has major potential for misuse as a performance-enhancing drug in several sports. Therefore, an efficient and selective separation method is required to detect and monitor the level of ATE in the body. This paper presents a molecularly imprinted polymer with specific and selective binding to ATE using precipitation polymerization. We show that when employed in an optimized molecular imprinted solid phase extraction (MI-SPE) protocol, recoveries of 93.65 ± 1.29% from spiked blood serum with excellent discrimination from other ß-blocker drugs is possible. The methodology used in this study includes molecular modeling interaction between ATE and itaconic acid (ITA) as functional monomer, followed by determination of binding constants with spectrophotometry, synthesis of the polymer using precipitation polymerization and ending with characterization and application of polymers to extract ATE in serum. Docking analysis revealed a binding affinity between ATE and ITA of -2.0 kcal/mol with the formation of hydrogen bonding. The association constant between ATE and ITA was studied by UV titration in two different solvents, with evidence of an association constant 6.277 × 102 M-1 measured in acetonitrile: methanol (1:1). An optimized MI-SPE protocol was developed for the extraction of ATE from spiked blood serum, obtaining recoveries of 93.65% with excellent selectivity toward other ß-blocker drugs.

Design and Optimization of Colorimetric Paper-Based Analytical Device for Rapid Detection of Allopurinol in Herbal Medicine.

Traditional herbal medicine in Indonesia is still in great demand and popular in society. The Indonesian government regulations state that herbal medicine should not contain chemical drug due to the toxic effect of uncontrolled consumption. Allopurinol is one of the drugs commonly added to herbal medicine for the treatment of chronic gout. Paper-based analytical device is one of the latest forms of analysis that has been widely used for the identification of chemical elements, environmental contamination, bacteria, and many more. In this study, experiments were conducted using Whatman filter paper No. 1, No. 2, and No. 4 and Whatman chromatography as a paper, and 9 colorimetric reagents were tested for allopurinol detection in herbal medicine. There were 5 specific reagents that reacted positively with allopurinol and only 3 reagents that can be applied to the paper, that is, Folin-Ciocalteu, Tollens, and p-DAB reagent. The results of the optimization show that the most optimal immersion time was 60 minutes with a drying time of 30 minutes at 50°C. Each filter paper has different characteristic; however, there was no significant difference when all of the papers were used as PAD for allopurinol detection.

Development and validation of simple simultaneous analysis for amlodipine and glibenclamide by nonderivatization high-performance liquid chromatography-fluorescence.

Studies have shown that about 65% of diabetics have hypertension. Treatment for diabetic patients with hypertension is usually given a combination of drugs such as amlodipine (AML) and glibenclamide (GLI). The aim of this study was to develop and validate the simple simultaneous analysis method for separation of AML and GLI using high-performance liquid chromatography (HPLC) with fluorescence detector without derivatization. The arrangement of isocratic and gradient methods, mobile phase compositions, and flow rates to develop and validate the simple simultaneous analysis method for separation of AML and GLI by nonderivatization HPLC fluorescence was done. Optimum condition was obtained using an RP 18 (125 mm × 4 mm, i.d., 5 µm) and guard column RP 18 (4 mm × 4 mm, i.d., 5 µm) with mobile phase composition containing acetonitrile and phosphate buffer pH 3.0 using a 20:80 gradient condition at flow rate 1.0 ml/min measured at 361 nm for λ excitation and 442 nm for λ emission for AML and 235 nm for λ excitation and 354 nm for λ emission for GLI. The analysis of AML and GLI demonstrated a valid result with r 2 value 0.999, recoveries were 100.04% and 99.14% relative standard deviations were 0.508% and 0,797%, respectively, detection limits were 0.055 and 0.104 µg/ml, and quantification limits were 0.166 and 0.316 µg/ml, respectively. An accurate method of separation for AML and GLI using HPLC with fluorescence detector without derivatization has been validated.