Page 86 - SDIR5 Abstract book 21 12 2021.

POSTER PRESENTATIONS

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Development and validation of a simple and reliable UV-coupled HPLC assay for the
determination of gemcitabine in serum: application in pharmacokinetic analysis

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1
Lafazanis Kostas , Begas Elias , Sakellaridis Nikos , Iatrou Hermes and Dimas Konstantinos
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1 Department of pharmacology, School of medicine, university of Thessaly, Larissa, Greece.
2 Industrial Chemistry Laboratory, Department of Chemistry, National and Kapodistrian University of Athens,
Athens, Greece

Background: Gemcitabine (gem) is an important chemotherapeutic drug used for the treatment of
pancreatic, and other cancers either as monotherapy or in combination with other medicines.
Aim: The development of a simple and reliable HPLC UV-coupled method for determining gemcitabine in
serum. Material and methods: Sample preparation consisted of a single protein precipitation step with
perchloric acid. Analysis was accomplished by a reversed-phase column eluted isocratically by sodium
phosphate buffer (pH 6.6) and methanol (97/3, v/v) at flow-rate 1mL/min, detection wavelength at 267nm
and column temperature at 40ºC. 1,7-dimethyluric acid was used as an internal standard. For the
pharmacokinetic study NOD/SCID mice (n= 5/group) were used and the drug was administered. Mice
received a single dose of gem at 100mg/kg either subcutaneous (sc) or intraperitoneal (ip). Blood samples
were collected at 5, 15, 30min and 1, 2, 4 and 6h post gem administration. Results: Duration of analysis
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was ~12.5min. Calibration curve was linear with r = 0.999 over the range 1-400μM, coefficient of variation
was <6.52% and bias <-7.77 %. Mean recovery of gem was 96.53% and the limit of detection was 0.17μM.
T1/2, Tmax, Cmax and AUC0-t were 1.03h, 0.083h, 272.14μmol/L and 135.99μmol/L*h for sc while the
respective values for ip administration were 0.85 h, 0.083 h, 291.542 μmol/L and 121.227μmol/L*h.
Conclusion: We developed a simple, valid and inexpensive HPLC method coupled to UV detection of
gemcitabine for the determination of the drug in serum which may also be suitable for clinical practice.
Keywords: gemcitabine, HPLC, pharmacokinetics
Acknowledgment: This research has been co-financed by the European Union and Greek national funds
through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call
RESEARCH – CREATE – INNOVATE (project code:T1EDK-01612)

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