Development and validation of indocyanine green assay in human plasma using High-Performance Liquid Chromatography with PDA detector

  • Sumith K Mathew Department of Pharmacology and Clinical Pharmacology, Clinical Pharmacology Unit, Christian Med-ical College, Vellore, Tamil Nadu – 632004, India
  • Blessed Winston A Department of Pharmacology and Clinical Pharmacology, Clinical Pharmacology Unit, Christian Med-ical College, Vellore, Tamil Nadu – 632004, India
  • Aswathy Mathew Department of Pharmacology and Clinical Pharmacology, Clinical Pharmacology Unit, Christian Med-ical College, Vellore, Tamil Nadu – 632004, India
  • Jeana Jacob Department of Pharmacology and Clinical Pharmacology, Clinical Pharmacology Unit, Christian Med-ical College, Vellore, Tamil Nadu – 632004, India
  • Ratna Prabha Department of Pharmacology and Clinical Pharmacology, Clinical Pharmacology Unit, Christian Med-ical College, Vellore, Tamil Nadu – 632004, India
  • Binu Susan Mathew Department of Pharmacology and Clinical Pharmacology, Clinical Pharmacology Unit, Christian Med-ical College, Vellore, Tamil Nadu – 632004, India

Abstract

A robust and economical assay for routine determination of indocyanine green pharmacokinetics was developed and validated using high-performance liquid chromatography with a photodiode array detector. Plasma specimens from critically ill patients and those with hepatitis on various co-medications were used as blanks for validation of this assay. Extraction of indocyanine green was performed by simple protein precipitation with acetonitrile, and the supernatant was separated using an octadecyl column with detection at 784 nm. Blanks were found to have no interference for 40 blanks of patients who were on 56 different medications. The precision for LLOQ (0.5 µg/ml) as determined by the percentage coefficient of variation was 1.19. Stability of plasma calibration standards and stock were determined over a period of 61 days, and ICG was found to be stable for 20 days. Stability of whole blood specimens containing ICG was determined at 4°C for a period of 4 hours.

Keywords: HPLC assay, Indocyanine green, Photodiode array detec-tor

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Published
2019-04-14
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How to Cite
Sumith K Mathew, Blessed Winston A, Aswathy Mathew, Jeana Jacob, Ratna Prabha, & Binu Susan Mathew. (2019). Development and validation of indocyanine green assay in human plasma using High-Performance Liquid Chromatography with PDA detector. International Journal of Research in Pharmaceutical Sciences, 10(2), 1007-1012. https://doi.org/10.26452/ijrps.v10i2.373
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