TY - JOUR

T1 - Distribution of platinum between nuclear and cytosolic fractions

T2 - can subcellular fractionation be performed quantitatively?

AU - Dam, Celina Støving

AU - Lambert, Ian Henry

AU - Gammelgaard, Bente

AU - Stürup, Stefan

PY - 2019

Y1 - 2019

N2 - The feasibility of quantitatively tracking platinum, derived from platinum-based compounds, during subcellular fractionation was studied. Cisplatin-exposed murine Ehrlich Lettré Ascites cells were fractionated into cytosolic and crude nuclear fractions. The latter was subsequently purified. Every residue and fraction produced during the fractionation procedure were collected and the platinum content determined by inductively coupled plasma mass spectrometry. Western blotting verified that the nuclear and cytosolic fractions were pure. It was found that 18% of platinum taken up by the cells was located in the nuclear fraction while 66% was located in the cytosolic fraction. Accumulated uncertainty originating from invariable sample characteristics and giving fraction purity priority had a negative effect on platinum recovery. Thus, overall 81% (n = 3, RSD = 3.4%) of the platinum taken up by the cells was recovered in the residues and final fractions. In conclusion, a reliable intracellular localization and quantitation of platinum following administration of Cisplatin can be determined by application of the method.

AB - The feasibility of quantitatively tracking platinum, derived from platinum-based compounds, during subcellular fractionation was studied. Cisplatin-exposed murine Ehrlich Lettré Ascites cells were fractionated into cytosolic and crude nuclear fractions. The latter was subsequently purified. Every residue and fraction produced during the fractionation procedure were collected and the platinum content determined by inductively coupled plasma mass spectrometry. Western blotting verified that the nuclear and cytosolic fractions were pure. It was found that 18% of platinum taken up by the cells was located in the nuclear fraction while 66% was located in the cytosolic fraction. Accumulated uncertainty originating from invariable sample characteristics and giving fraction purity priority had a negative effect on platinum recovery. Thus, overall 81% (n = 3, RSD = 3.4%) of the platinum taken up by the cells was recovered in the residues and final fractions. In conclusion, a reliable intracellular localization and quantitation of platinum following administration of Cisplatin can be determined by application of the method.

KW - Cisplatin

KW - Inductively coupled plasma mass spectrometry

KW - Quantitative

KW - Subcellular distribution

KW - Subcellular fractionation

U2 - 10.1016/j.jpba.2018.11.046

DO - 10.1016/j.jpba.2018.11.046

M3 - Journal article

C2 - 30508755

AN - SCOPUS:85057452962

VL - 165

SP - 82

EP - 89

JO - Journal of Pharmaceutical and Biomedical Analysis

JF - Journal of Pharmaceutical and Biomedical Analysis

SN - 0731-7085

ER -