GB/T 45238-2025化學(xué)品用虹鱒肝S9亞細(xì)胞組分測定體外固有清除率試驗

ICS13.300

CCSA80

中華人民共和國國家標(biāo)準(zhǔn)

GB/T45238—2025

化學(xué)品用虹鱒肝S9亞細(xì)胞組分測定

體外固有清除率試驗

Chemicals—Determinationofinvitrointrinsicclearanceusingrainbowtrout

liverS9sub?cellularfraction（RT?S9）

2025?01?24發(fā)布2025?05?01實施

國家市場監(jiān)督管理總局

國家標(biāo)準(zhǔn)化管理委員會發(fā)布

GB/T45238—2025

目次

前言··························································································································Ⅲ

引言··························································································································Ⅳ

1范圍·······················································································································1

2規(guī)范性引用文件········································································································1

3術(shù)語和定義、符號和縮略語··························································································1

4試驗原理·················································································································3

5受試物信息··············································································································3

6參比物····················································································································4

7試驗準(zhǔn)備·················································································································4

8試驗程序·················································································································5

9數(shù)據(jù)處理·················································································································6

10質(zhì)量保證與質(zhì)量控制·······························································································7

11結(jié)果報告···············································································································7

附錄A（資料性）虹鱒肝S9亞細(xì)胞組分的制備···································································9

附錄B（資料性）虹鱒肝亞細(xì)胞組分的表征·······································································16

附錄C（資料性）酶促失活的虹鱒肝S9亞細(xì)胞組分的制備···················································17

附錄D（資料性）初步試驗反應(yīng)條件的建立·······································································18

附錄E（資料性）試驗方法的建立···················································································20

附錄F（資料性）緩沖溶液、輔因子、丙甲甘肽溶液的配制和RT?S9的稀釋······························22

參考文獻(xiàn)····················································································································25

Ⅰ

GB/T45238—2025

前言

本文件按照GB/T1.1—2020《標(biāo)準(zhǔn)化工作導(dǎo)則第1部分：標(biāo)準(zhǔn)化文件的結(jié)構(gòu)和起草規(guī)則》的規(guī)

定起草。

請注意本文件的某些內(nèi)容可能涉及專利。本文件的發(fā)布機(jī)構(gòu)不承擔(dān)識別專利的責(zé)任。

本文件由全國危險化學(xué)品管理標(biāo)準(zhǔn)化技術(shù)委員會（SAC/TC251）提出并歸口。

本文件起草單位：廣東省科學(xué)院微生物研究所（廣東省微生物分析檢測中心）、生態(tài)環(huán)境部固體廢

物與化學(xué)品管理技術(shù)中心、中檢科健（天津）檢驗檢測有限責(zé)任公司、中國水產(chǎn)科學(xué)研究院珠江水產(chǎn)研

究所。

本文件主要起草人：梅承芳、許玫英、陳桂蘭、王青柏、陳會明、竇從從、林健輝、曾國驅(qū)、周麗麗、

劉曉建、梁秋、柳燕貞、劉純新、馮寶欣、譚文蔚、區(qū)麗嫻、梁平緩、王英英。

Ⅲ

GB/T45238—2025

引言

為提高化學(xué)品對魚類生物蓄積作用的預(yù)測能力，需建立魚體內(nèi)肝臟生物轉(zhuǎn)化的評估方法并將相關(guān)

信息納入已建立的計算機(jī)模型。目前主要利用肝組織的體外代謝系統(tǒng)測定化學(xué)品的固有清除率。

本文件描述了基于虹鱒魚（Oncorhynchusmykiss）肝S9亞細(xì)胞組分（RT?S9），采用底物清除法測定

受試物體外固有清除率（）的試驗方法。OECD化學(xué)品測試導(dǎo)則No.319A和OECD化學(xué)

CLINVITRO，INT

品測試指南文件No.280描述了開展試驗的詳細(xì)步驟，以及如何將用于魚類生物蓄積性的

CLINVITRO，INT

計算機(jī)模型預(yù)測。

簡而言之，根據(jù)確定的估算肝臟固有清除率，并采用基于生理學(xué)的毒代動力學(xué)

CLINVITRO，INT

（PBTK）模型進(jìn)一步評估魚類的生物蓄積作用。也可采用合適的體外-體內(nèi)外推模型（IVIVE）將該值

外推獲得全身（體內(nèi)）的生物轉(zhuǎn)化速率常數(shù)。如果受試物處于模型的應(yīng)用范圍內(nèi)，可將測得的

作為輸入信息，應(yīng)用于基于生物轉(zhuǎn)化的生物富集計算機(jī)預(yù)測模型。

CLINVITRO，INT

體內(nèi)生物轉(zhuǎn)化率可用于計算機(jī)模型預(yù)測生物富集系數(shù)（BCF）。將體外方法獲得的生物轉(zhuǎn)化數(shù)據(jù)

整合到魚類一室模型中，可顯著改善模型性能，與代謝缺失時得到的預(yù)測結(jié)果相比，整合生物轉(zhuǎn)化數(shù)據(jù)

后的模型對生物富集結(jié)果的預(yù)測更接近試驗值。

本文件的重復(fù)性已通過6個實驗室開展的比對試驗（針對5種受試物和1種參比物）進(jìn)行了驗證。

已對部分化學(xué)品開展了基于魚類肝細(xì)胞、肝S9亞細(xì)胞組分或微粒體的相關(guān)測試，并獲得了

。歐盟委員會聯(lián)合研究中心已開發(fā)魚類體外生物轉(zhuǎn)化數(shù)據(jù)庫，涵蓋了采用體外方法（肝細(xì)

CLINVITRO，INT

胞、S9、微粒體）測定的固有清除率數(shù)據(jù)，這些均為本文件的適用性提供了參考。

如采用本文件對混合物、難測試物質(zhì)（如不穩(wěn)定物質(zhì)）或適用范圍內(nèi)未指明的受試物開展測試，先

考慮試驗結(jié)果是否具有科學(xué)意義。

Ⅳ

GB/T45238—2025

化學(xué)品用虹鱒肝S9亞細(xì)胞組分測定

體外固有清除率試驗

1范圍

本文件描述了用虹鱒肝S9亞細(xì)胞組分測定化學(xué)品體外固有清除率的試驗方法，包括試驗原理、受

試物信息、參比物、試驗準(zhǔn)備、試驗程序、數(shù)據(jù)處理、質(zhì)量保證與質(zhì)量控制和結(jié)果報告。

本文件適用于測定化學(xué)品的體外固有清除率，可準(zhǔn)確定量的最低體外一級清除速率常數(shù)（Ke）約為

0.05h-1~0.14h-1。如有特殊需要，也可用于識別代謝產(chǎn)物。

2規(guī)范性引用文件

下列文件中的內(nèi)容通過文中的規(guī)范性引用而構(gòu)成本文件必不可少的條款。其中，注日期的引用文

件，僅該日期對應(yīng)的版本適用于本文件；不注日期的引用文件，其最新版本（包括所有的修改單）適用于

本文件。

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