Validation of an HPLC method for estimation of cefotaxime from dried blood spot: alternative to plasma-based PK evaluation in neonates

Aim: Pharmacokinetic evaluation of cefotaxime in neonates is currently a challenge due to the large volume requirement of blood for its analysis by existing methods. A dried blood spot (DBS) based method is the best alternative. Materials & methods: We validated an HPLC method for estimation of cefotaxime from DBS and plasma. Extraction employed a simple procedure using acetonitrile and buffer. Selective separation of cefotaxime was achieved on a C8 column using gradient programming. Results & conclusion: The linearity of the method ranged from 2 to 200 μg/ml with acceptable precision and accuracy for both plasma and DBS. Hematocrit was not affecting the assay accuracy. A strong correlation and interchangeability observed with the plasma method proves its clinical validity for application to PK evaluations.

Download Full-text

Optimization of an HPLC method for phenylalanine and tyrosine quantization in dried blood spot

Clinical Biochemistry ◽

10.1016/j.clinbiochem.2013.08.022 ◽

2013 ◽

Vol 46 (18) ◽

pp. 1892-1895 ◽

Cited By ~ 12

Author(s):

Rita Pecce ◽

Emanuela Scolamiero ◽

Laura Ingenito ◽

Giancarlo Parenti ◽

Margherita Ruoppolo

Keyword(s):

Hplc Method ◽

Dried Blood Spot ◽

Blood Spot

Download Full-text

Dried Blood Spot Screening Can Identify Thyroid Disorders in Pregnant Women

Clinical Endocrinology News ◽

10.1016/s1558-0164(06)70448-5 ◽

2006 ◽

Vol 1 (12) ◽

pp. 24

Author(s):

Heidi Splete

Keyword(s):

Pregnant Women ◽

Dried Blood Spot ◽

Thyroid Disorders ◽

Blood Spot

Download Full-text

RNA-Seq of Dried Blood Spot Samples using Lexogen’s QuantSeq 3’ mRNA-Seq FWD assay

10.26226/morressier.5ebd45acffea6f735881b1a1 ◽

2020 ◽

Author(s):

Steve Cole

Keyword(s):

Dried Blood Spot ◽

Rna Seq ◽

Blood Spot

Download Full-text

QUANTITATIVE DETERMINATION OF CURCUMIN IN TURMERIC POWDER AND TURMERIC–HONEY BALL PILLS BY HPLC

Journal of Medicine and Pharmacy ◽

10.34071/jmp.2018.4.6 ◽

2018 ◽

Vol 8 (4) ◽

pp. 42-47

Author(s):

Tien Nguyen Huu ◽

Tram Le Thi Bao ◽

Ngoc Nguyen Thi Nhu ◽

Thang Phan Phuoc ◽

Khan Nguyen Viet

Keyword(s):

Acetic Acid ◽

Gastric Mucosa ◽

Quantitative Determination ◽

Mobile Phase ◽

Curcuma Longa ◽

Biological Marker ◽

Hplc Method ◽

Chromatography Analysis ◽

Precision And Accuracy

Background: Curcumin is a major ingredient in turmeric (Curcuma longa L., Zingiberaceae), which has important activities such as anti-tumor, anti-inflammatory, antioxidant, anti-ischemia, protection of gastric mucosa etc,. Curcumin can be considered as a biological marker of turmeric and turmeric products. Objectives: Developing an HPLC method for quantification of curcumin in turmeric powder and turmeric - honey ball pills; applying this method for products on the market. Materials and methods: turmeric powder and turmeric - honey ball pills collected in Thua Thien Hue province. After optimization process, the method was validated and applied to evaluate the content of curcumin. Results: The chromatography analysis was performed with: Zorbaz Eclipse XDB-C18 (150 × 4.6 nm; 5 µm); Mobile phase: acetonitril: 2% acetic acid (45:55), Flow rate was kept constant at 1.0 ml/min; Detector PDA (420 nm). The method was validated for the HPLC system compatibility, specificity, linearity range, precision and accuracy; the recovery greater than 98%. Conclusion: The developed HPLC method can determine curcumin in turmeric powder and turmeric - honey ball pills. Key words: Curcumin, turmeric powder, turmeric-honey ball pills, quantitative determination, HPLC

Download Full-text

Development, Validation and Application of HPLC Method for Metformin in Rabbit Plasma

Current Pharmaceutical Analysis ◽

10.2174/1573412914666180308124858 ◽

2019 ◽

Vol 15 (6) ◽

pp. 574-579

Author(s):

Muhammad Ubaid ◽

Mahmood Ahmad ◽

Farhan Ahmad Khan ◽

Ghulam Murtaza

Keyword(s):

Flow Rate ◽

Present Method ◽

Mobile Phase ◽

Hplc Method ◽

Plasma Samples ◽

Rabbit Plasma ◽

Uv Detector ◽

Reverse Phase ◽

T Values ◽

Pharmacokinetic Evaluation

Objective:This study was aimed at conducting a pharmacokinetic evaluation of metformin in rabbit plasma samples using rapid and sensitive HPLC method and UV detection.Methods:Acetonitrile was used for protein precipitation in the preparation of plasma samples. Reverse phase chromatography technique with silica gel column (250 mm × 4.6 mm, 5 μm) at 30°was used for the separation purpose. Methanol and phosphate buffer (pH 3.2) mixture was used as a mobile phase with flow rate 0.8 ml/min. The wavelength of UV detector was adjusted at 240 nm.Results:The calibration curve was linear in a range of 0.1-1 µg/ml with R² = 0.9982. The precision (RSD, %) values were less than 2%, whereas, accuracy of method was higher than 92.37 %. The percentage recovery values ranged between 90.14 % and 94.97 %. LOD and LOQ values were 25 ng/ml and 60 ng/ml, respectively. Cmax and AUC0-t values were found to be 1154.67 ± 243.37 ng/ml and 7281.83 ± 210.84 ng/ml.h, respectively after treating rabbits with a formulation containing 250 mg metformin.Conclusion:Based on the above findings, it can be concluded that present method is simple, precise, rapid, accurate and specific and thus, can be efficiently used for the pharmacokinetic study of metformin.

Download Full-text

Fabry Cardiomyopathy: Current Practice and Future Directions

Cells ◽

10.3390/cells10061532 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1532

Author(s):

Jeffrey Yim ◽

Olivia Yau ◽

Darwin F. Yeung ◽

Teresa S. M. Tsang

Keyword(s):

Fabry Disease ◽

Early Stage ◽

Point Of Care ◽

Left Ventricular ◽

The Body ◽

Cardiac Biomarkers ◽

Dried Blood Spot ◽

Point Of Care Ultrasound ◽

Enzyme Replacement ◽

Blood Spot

Fabry disease (FD) is an X-linked lysosomal storage disorder caused by mutations in the galactosidase A (GLA) gene that result in deficient galactosidase A enzyme and subsequent accumulation of glycosphingolipids throughout the body. The result is a multi-system disorder characterized by cutaneous, corneal, cardiac, renal, and neurological manifestations. Increased left ventricular wall thickness represents the predominant cardiac manifestation of FD. As the disease progresses, patients may develop arrhythmias, advanced conduction abnormalities, and heart failure. Cardiac biomarkers, point-of-care dried blood spot testing, and advanced imaging modalities including echocardiography with strain imaging and magnetic resonance imaging (MRI) with T1 mapping now allow us to detect Fabry cardiomyopathy much more effectively than in the past. While enzyme replacement therapy (ERT) has been the mainstay of treatment, several promising therapies are now in development, making early diagnosis of FD even more crucial. Ongoing initiatives involving artificial intelligence (AI)-empowered interpretation of echocardiographic images, point-of-care dried blood spot testing in the echocardiography laboratory, and widespread dissemination of point-of-care ultrasound devices to community practices to promote screening may lead to more timely diagnosis of FD. Fabry disease should no longer be considered a rare, untreatable disease, but one that can be effectively identified and treated at an early stage before the development of irreversible end-organ damage.

Download Full-text

Use of dried blood spot samples for SARS-CoV-2 antibody detection using the Roche Elecsys ® high throughput immunoassay

Journal of Clinical Virology ◽

10.1016/j.jcv.2021.104739 ◽

2021 ◽

Vol 136 ◽

pp. 104739

Author(s):

Ranya Mulchandani ◽

Ben Brown ◽

Tim Brooks ◽

Amanda Semper ◽

Nicholas Machin ◽

...

Keyword(s):

High Throughput ◽

Dried Blood Spot ◽

Antibody Detection ◽

Blood Spot

Download Full-text

A surrogate virus neutralization test to quantify antibody-mediated inhibition of SARS-CoV-2 in finger stick dried blood spot samples

Scientific Reports ◽

10.1038/s41598-021-94653-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Amelia E. Sancilio ◽

Richard T. D’Aquila ◽

Elizabeth M. McNally ◽

Matthew P. Velez ◽

Michael G. Ison ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Venous Blood ◽

Host Cells ◽

Dried Blood Spot ◽

Blood Collection ◽

Widespread Application ◽

Live Virus ◽

Dilution Series ◽

Wide Range ◽

Blood Spot

AbstractThe spike protein of SARS-CoV-2 engages the human angiotensin-converting enzyme 2 (ACE2) receptor to enter host cells, and neutralizing antibodies are effective at blocking this interaction to prevent infection. Widespread application of this important marker of protective immunity is limited by logistical and technical challenges associated with live virus methods and venous blood collection. To address this gap, we validated an immunoassay-based method for quantifying neutralization of the spike-ACE2 interaction in a single drop of capillary whole blood, collected on filter paper as a dried blood spot (DBS) sample. Samples are eluted overnight and incubated in the presence of spike antigen and ACE2 in a 96-well solid phase plate. Competitive immunoassay with electrochemiluminescent label is used to quantify neutralizing activity. The following measures of assay performance were evaluated: dilution series of confirmed positive and negative samples, agreement with results from matched DBS-serum samples, analysis of results from DBS samples with known COVID-19 status, and precision (intra-assay percent coefficient of variation; %CV) and reliability (inter-assay; %CV). Dilution series produced the expected pattern of dose–response. Agreement between results from serum and DBS samples was high, with concordance correlation = 0.991. Analysis of three control samples across the measurement range indicated acceptable levels of precision and reliability. Median % surrogate neutralization was 46.9 for PCR confirmed convalescent COVID-19 samples and 0.1 for negative samples. Large-scale testing is important for quantifying neutralizing antibodies that can provide protection against COVID-19 in order to estimate the level of immunity in the general population. DBS provides a minimally-invasive, low cost alternative to venous blood collection, and this scalable immunoassay-based method for quantifying inhibition of the spike-ACE2 interaction can be used as a surrogate for virus-based assays to expand testing across a wide range of settings and populations.

Download Full-text