METHODS AND APPARATUS FOR EXTRACTING DOXORUBICIN FROM BLOOD AND MEASURING DOXORUBICIN IN BLOOD
Doxorubicin is extracted from blood using anionic material, such as a resin comprising sulfonated polystyrene divinylbenzene beads, and polyethersulfone membrane, or both. After exposing the resin and/or membrane to blood in order to remove doxorubicin therefrom, the doxorubicin maybe extracted from the resin and/or membrane by exposing the material to an extraction solution, sonicating the extraction solution to enhance release of the doxorubicin, and repeating the exposure and sonication in order to remove substantially all of doxorubicin from the resin.
1 . A method for extracting doxorubicin from a patient's blood, said method comprising:
exposing an anionic material to the patient's blood, said anionic material being capable of binding doxorubicin; and
extracting doxorubicin from the anionic material by:
(a) placing the exposed anionic material into an extraction solution in the presence of a surfactant;
(b) sonicating the extraction solution to release doxorubicin into the extraction solution;
(c) separating the anionic material from the extraction solution to produce a supernatant including the extracted doxorubicin;
(d) resuspending the separated anionic material in fresh extraction solution; and
(e) sonicating the fresh extraction solution to release additional doxorubicin into the fresh extraction solution.
2 . A method as in claim 1 , wherein the sulfonated anionic material is selected from the group consisting of polystyrene divinylbenzene beads, a polyethersulfone sulfonate membrane, and a nucleic acid-derivatized filter material.
3 . A method as in claim 1 , wherein the step of exposing the anionic material comprises introducing the anionic material into the patient's vasculature in vivo.
4 . A method as in claim 1 , wherein introducing the anionic material comprises intravascularly positioning a catheter which carries the anionic material into the vasculature.
5 . A method as in claim 1 , wherein the extraction solution comprises an aqueous KCl solution combined with an organic phase.
6 . A method as in claim 5 , wherein the organic phase comprises a mixture of acetone, ethanol, and acetonitrile.
7 . A method as in claim 1 , wherein the extraction solution comprises from 1:1.5 to 1.5:1 parts aqueous KCl solution and organic phase by volume.
8 . A method as in claim 5 , wherein the surfactant comprises from 0.5% to 2% by volume of a surfactant selected from the group consisting of CHAPS, sodium docecyl sulfate, Triton X-100, and CTAB.
9 . A method as in claim 1 , wherein sonicating comprises placing the extraction solution with the anionic material and the surfactant in an ultrasonic bath.
10 . A method as in claim 1 , wherein sonicating comprises immersing a sonication probe into the extraction solution.
11 . A method as in claim 1 , wherein the sonicating step is performed for 15 minutes to 1 hour.
12 . A method as in claim 1 , wherein the sonicating step is performed in two steps with a supernatant being separated, the resin placed in a fresh extraction solution, and the sonicating step repeated, wherein a first supernatant and a second supernatant are created.
13 . A method as in claim 1 , wherein from 80% to 95% of the doxorubicin is extracted in the first sonication and the remaining 5% to 20% is removed in the second sonication, with a total removal of 99% to 100%.
14 - 23 . (canceled)