title: Polydiacetylene Liposomal Aequorin Bioluminescent Device for Detection of Hydrophobic Compounds.
abstract: In this study, a polydiacetylene liposomal aequorin bioluminescent device (PLABD) that functioned through control of the membrane transport of Ca(2+) ions was developed for detecting hydrophobic compounds. In the PLABD, aequorin was encapsulated in an internal water phase and a calcium ionophore (CI) was contained in a hydrophobic region. Membrane transport of Ca(2+) ions across the CI was suppressed by polymerization between diacetylene molecules. On addition of an analyte, the membrane transport of Ca(2+) ions across the CI increased, and Ca(2+) ions from the external water phase could diffuse into the internal water phase via the CI, which resulted in bioluminescence of the aequorin. Lidocaine, procaine, and procainamide were used as model compounds to test the validity of the detection mechanism of the PLABD. When each analyte was added to a suspension of the PLABD, bioluminescence from the aequorin in the PLABD was observed, and the level of this bioluminescence increased with increasing analyte concentration. There was a linear relationship between the logarithm of the analyte concentration and the bioluminescence for all analytes as follows: R = 0.89 from 10 nmol L(-1) to 10 mmol L(-1) for lidocaine, R = 0.66 from 10 nmol L(-1) to 100 μmol L(-1) for procaine, and R = 0.74 from 100 nmol L(-1) to 100 μmol L(-1) for procainamide. Compared to the traditional colorimetric method using polydiacetylene liposome, the PLABD was superior for both the sensitivity and dynamic range. Thus, PLABD is a valid, simple, and sensitive signal generator for detection of hydrophobic compounds that interact with PLABD membranes.
authors.affiliation: Department of Analytical Chemistry, Kyoto Pharmaceutical University , 5 Nakauchicho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Yamamoto
authors.affiliation: Department of Analytical Chemistry, Kyoto Pharmaceutical University , 5 Nakauchicho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Shigehiko
authors.initials: S
authors.lastname: Takegami
authors.affiliation: Department of Analytical Chemistry, Kyoto Pharmaceutical University , 5 Nakauchicho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Konishi
authors.affiliation: Department of Analytical Chemistry, Kyoto Pharmaceutical University , 5 Nakauchicho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Hikari
authors.initials: H
authors.lastname: Horikawa
authors.affiliation: Department of Analytical Chemistry, Kyoto Pharmaceutical University , 5 Nakauchicho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Sayumi
authors.initials: S
authors.lastname: Yonezawa
authors.affiliation: Department of Analytical Chemistry, Kyoto Pharmaceutical University , 5 Nakauchicho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Kitade
mesh_terms: D000331:Aequorin / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D008012:Lidocaine / Q000032:analysis*; D008081:Liposomes / Q000737:chemistry; D008163:Luminescent Measurements; D015394:Molecular Structure; D000078789:Polyacetylene Polymer; D011108:Polymers / Q000737:chemistry*; D053279:Polyynes / Q000737:chemistry*; D011342:Procainamide / Q000032:analysis*; D011343:Procaine / Q000032:analysis*
chemical_list: D008081:Liposomes; D000078789:Polyacetylene Polymer; D011108:Polymers; D053279:Polyynes; C082361:polydiacetylene; D011343:Procaine; D000331:Aequorin; D008012:Lidocaine; D011342:Procainamide; D002118:Calcium