@article{oai:toyama.repo.nii.ac.jp:00005249,
 author = {ZHAO, Jing and SUN, Xiao-Fei and Nakamura, Norio and ZUO, Feng and YANG, Xiu-Wei and Akao, Teruaki and Hattori, Masao},
 issue = {5},
 journal = {和漢医薬学雑誌 = Journal of traditional medicines},
 month = {Oct},
 note = {application/pdf, For the purpose of quantitative determination of mesaconitine, an extremely toxic and major alkaloid in aconite tuber, a highly sensitive enzyme immunoassay (EIA) was developed as follows; Firstly, a hapten [N-desethylaconitine N-glutarate (DEAG)] was synthesized by introduction of a glutaryl group to the nitrogen atom of aconitine after elimination of an N-ethyl group. Subsequently it was coupled with bovine serum albumin (BSA) and β-galactosidase (β-Gal) to give animmunogen (DEAG-BSA) and a labeled antigen (DEAG-β-Gal), respectively. After immunization of female albino rabbits with DEAG-BSA for six months to elicit a polyclonal antiserum (As-DEAG), the experimental conditions were optimized for a highly sensitive EIA. At 10^3-fold dilution of DEAG-β-Gal and 2.5 × 10^4-fold dilution of As-DEAG, the assay exhibited a linear range of 0.005-5 ng/tube for mesaconitine. Under the same conditions, the antiserum had a 24.4 % cross-reaction with aconitine and virtually no reaction with benzoylmesaconine. Next, the EIA method was applied to the disposition study on mesaconitine to monitor the plasma concentration after oral administration of mesaconitine at 1 mg/kg to rats, its low bioavailability being suggested from Cmax and AUC values obtained.
 附子に含まれるaconitineと並ぶ猛毒なアルカロイドであるmesaconitineの定量の目的で,高感度酵素免疫測定法(EIA)を開発した。すなわち,最初にaconitineのN-エチルグループを除去した後,二級窒素原子にグルタール酸基をアミドとして導入することによりハプテンを合成した。次いでこのハプテンとN-ヒドロキシスクシニイミドを反応させ活性エステルを合成した後,仔ウシ血清アルブミン(BSA)あるいはβ-ガラクトシダーゼ(β-Gal)と反応させ,それぞれ免疫原(DEAG-BSA),標識抗原(DEAG-β-Gal)を作製した。DEAG-BSAを白色ウサギに6ヶ月間投与し,抗血清(As-DEAG)を得た後,高感度EIAを確立するために諸条件を検討した。10^3倍希釈したDEAG-β-Galと2.5×10^4倍希釈したAs-DEAG抗血清を用いた場合,mesaconitineに対して0.005～5ng/の濃度範囲で直線性が示された。同条件下,この抗血清はaconitineと24.4%の交差反応を示したが,benzoylmesaconineに対しては交差反応を示さなかった。このEIAをmesaconitineの薬物動態研究に応用した結果, 1mg/kgのmesaconitineを経口投与した場合の血清中の濃度測定が可能であり,猛毒性アルカロイドであるmesaconitineの動態パラメーターを初めて得ることができた。, Article},
 pages = {201--207},
 title = {Development of an enzyme immunoassay system for mesaconitine and its application to the disposition study on mesaconitine},
 volume = {20},
 year = {2003}
}