Bardoxolone Methyl

别名: RTA 402, TP-155, NSC 713200, CDDO Methyl Ester, CDDO-Me 中文名称：甲基巴多索隆

目录号：S8078 Purity: 99.86%

Bardoxolone Methyl (RTA 402, TP-155, NSC 713200, CDDO Methyl Ester, CDDO-Me) 是一种IKK抑制剂，具有强的促凋亡和抗炎活性。同时还是有效的Nrf2激活剂和NF-κB抑制剂。Bardoxolone Methyl 可抑制铁死亡。Bardoxolone Methyl 在癌细胞中可诱导凋亡和自噬。

Bardoxolone Methyl Chemical Structure

CAS: 218600-53-4

规格	价格	库存
25mg	RMB 814.73	现货
200mg	RMB 4656.69	现货
1g	RMB 12700	现货
更大包装有超大折扣
400-668-6834 info@selleck.cn

客户使用Selleck的Bardoxolone Methyl发表文献28篇

客户使用该产品的3个实验数据

产品质控

批次：纯度： 99.86%

99.86

Bardoxolone Methyl相关产品

相关靶点	IκB IKK	点击展开
相关化合物库	FDA药物库免疫/炎症分子化合物库小分子免疫肿瘤化合物库 NF-κB信号通路库 FDA抗癌药物库	点击展开

IκB/IKK抑制剂选择性比较

细胞实验数据示例

细胞系	实验类型	给药浓度	孵育时间	活性描述	文献信息
RAW264.7	Antioxidant assay	100 nM	18 hrs	Antioxidant activity in mouse RAW264.7 cells assessed as inhibition of tBHP-induced ROS production at 100 nM pretreated for 18 hrs before challenge measured after 15 mins by H2DCFA-based flow cytometry	24388806
PANC1343	Antiproliferative assay	300 to 1000 nM	72 hrs	Antiproliferative activity against mouse PANC1343 cells at 300 to 1000 nM after 72 hrs by MTT assay	24388806
BMDM	Antiinflammatory assay	0.5 uM	1 hr	Antiinflammatory activity in C57BL/6 mouse BMDM cells assessed as inhibition of LPS-stimulated TNFalpha production at 0.5 uM pretreated for 1 hr before LPS challenge after 8 to 24 hrs by immunoassay	22533790
HCT8	Function assay	1 uM	24 hrs	Inhibition of HIF-1alpha protein expression in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of HIF-1alpha protein expression in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of STAT3 protein phosphorylation in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of STAT3 protein phosphorylation in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of AKT protein phosphorylation in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of AKT protein phosphorylation in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of ERK protein phosphorylation in human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Function assay	1 uM	24 hrs	Inhibition of ERK protein phosphorylation in FU-resistant human HCT8 cells at 1 uM incubated for 24 hrs by Western blotting method	25675144
HCT8	Antiproliferative assay	1 uM	72 hrs	Antiproliferative activity against human HCT8 cells assessed as inhibition of cell proliferation at 1 uM after 72 hrs by MTT assay	25675144
HCT8	Antiproliferative assay	1 uM	72 hrs	Antiproliferative activity against 5-FU resistant human HCT8 cells assessed as inhibition of cell proliferation at 1 uM after 72 hrs by MTT assay	25675144
BEAS2B	Function assay	10 uM	6 hrs	Activation of Nrf2 in human BEAS2B cells assessed as increase in HO1 gene expression at 10 uM incubated for 6 hrs by qPCR method	26278028
H42E	Function assay	0.01 to 30 nM	1 hr	Stabilization of NRF2 in rat H42E cells expressing ARE8L at 0.01 to 30 nM after 1 hr by Western blot analysis	26908173
NHBE	Cytoprotective assay	0.001 to 0.1 uM	18 hrs	Cytoprotective activity in NHBE cells assessed as inhibition of tBHP-induced GSH depletion at 0.001 to 0.1 uM preincubated for 18 hrs followed by tBHP addition for 4 hrs by thiostar dye based fluorescence assay	27031670
NHBE	Function assay	100 nM	24 hrs	Inhibition of KEAP1/NRF2 interaction in NHBE cells assessed as increase in GCLM mRNA expression at 100 nM incubated for 24 hrs in presence of non targeting siRNA by qRT-PCR method	27031670
NHBE	Function assay	100 nM	24 hrs	Inhibition of KEAP1/NRF2 interaction in NHBE cells assessed as increase in NQO1 mRNA expression at 100 nM incubated for 24 hrs in presence of non targeting siRNA by qRT-PCR method	27031670
NHBE	Function assay	100 nM	48 hrs	Inhibition of KEAP1/NRF2 interaction in NHBE cells assessed as induction of NQO1 specific activity at 100 nM incubated for 48 hrs in presence of non targeting siRNA by MTT reduction assay	27031670
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as TNFalpha-induced NFkappaB activation at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by NFkappaB-driven luciferase reporter gene assay	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of iNOS mRNA expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by quantitative RT-PCR an	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of COX2 mRNA expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by quantitative RT-PCR an	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of MCP1 mRNA expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by quantitative RT-PCR an	28994286
intraglomerular mesangial cells	Function assay	0.65 mg/kg	12 weeks	Renoprotective activity in db/db mouse assessed as increase in number of intraglomerular mesangial cells at 0.65 mg/kg, ip administered trice per week for 12 consecutive weeks measured at 11 weeks post dose by H/E-staining based microscopic analysis	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of COX2 protein expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot method	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of iNOS protein expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot method	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as mitigation of TNFalpha-induced increase in ratio of nuclear to cytosolic p65 at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot	28994286
HEK293	Function assay	200 to 1000 nM	24 hrs	Inhibition of IKKbeta (unknown origin) transfected in HEK293 cells assessed as reduction in TNFalpha-induced upregulation of MCP1 protein expression at 200 to 1000 nM administered 6 hrs after TNFalpha stimulation measured for 24 hrs by Western blot method	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as upregulation of HO-1 mRNA expression at 200 to 1000 nM after 48 hrs by quantitative RT-PCR analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as upregulation of NQO1 mRNA expression at 200 to 1000 nM after 48 hrs by quantitative RT-PCR analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as activation of Nrf2 at 200 to 1000 nM after 48 hrs by ARE-driven luciferase reporter gene assay	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as increase in nuclear to cytosolic Nfr2 ratio at 200 to 1000 nM after 48 hrs by Western blot analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as increase in cytosolic HO-1 levels at 200 to 1000 nM after 48 hrs by Western blot analysis	28994286
HEK293	Function assay	200 to 1000 nM	48 hrs	Inhibition of Keap1/Nrf2 (unknown origin) interaction transfected in HEK293 cells assessed as increase in cytosolic NQO1 levels at 200 to 1000 nM after 48 hrs by Western blot analysis	28994286
A549/TR	Function assay	2.4 to 9.6 uM	24 hrs	Induction of ROS generation in human A549/TR cells at 2.4 to 9.6 uM after 24 hrs by DCFH-DA dye-based flow cytometric analysis	29501947
A549/TR	Function assay	4.8 uM	24 hrs	Downregulation of Lon expression in human A549/TR cells at 4.8 uM after 24 hrs by Western blot analysis	29501947
B16F10	Cytotoxicity assay		48 hrs	Cytotoxicity against mouse B16F10 cells after 48 hrs by MTT assay, IC50=5.85μM	24685545
HepG2	Cytotoxicity assay		48 hrs	Cytotoxicity against human HepG2 cells after 48 hrs by MTT assay, IC50=4.99μM	24685545
BMDM	Cytotoxicity assay		24 hrs	Cytotoxicity against C57BL/6 mouse BMDM cells assessed as LDH release after 24 hrs, MNTD=0.5μM	22533790
CCD-841-CoN	Antiproliferative assay		72 hrs	Antiproliferative activity against human CCD-841-CoN cells assessed as inhibition of cell proliferation after 72 hrs by MTT assay, IC50=0.316μM	25675144
HCT8	Antiproliferative assay		72 hrs	Antiproliferative activity against 5-FU resistant human HCT8 cells assessed as inhibition of cell proliferation after 72 hrs by MTT assay, IC50=0.363μM	25675144
HCT8	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT8 cells assessed as inhibition of cell proliferation after 72 hrs by MTT assay, IC50=0.399μM	25675144
H42E	Function assay		24 hrs	Induction of NRF2 activation in rat H42E cells expressing ARE8L assessed as reporter transgene activity after 24 hrs by luminescence assay, CD=0.0005μM	26908173
H42E	Cytotoxicity assay		24 hrs	Cytotoxicity against rat H42E cells expressing ARE8L assessed as cellular ATP level after 24 hrs by Celltiter-Glo luminescent cell viability assay, IC50=1.4μM	26908173
HaCaT-ARE-luc	Function assay		6 hrs	Activation of Nrf2 (unknown origin) expressed in human HaCaT-ARE-luc cells after 6 hrs by luciferase reporter gene assay, EC50=0.06μM	28753294
NIH/3T3	Function assay		6 hrs	Inhibition of TNF-alpha stimulated NF-kappaB (unknown origin) expressed in mouse NIH/3T3 cells after 6 hrs by luciferase reporter gene assay, IC50=1.2μM	28753294
HeLa	Function assay		6 hrs	Inhibition of IFN-gamma stimulated STAT3 (unknown origin) expressed in human HeLa cells after 6 hrs by luciferase reporter gene assay, IC50=2.38μM	28753294
HEK293	Cytotoxicity assay		24 hrs	Cytotoxicity against HEK293 cells assessed as reduction in cell viability after 24 hrs by MTT assay, IC50=2.2μM	28994286
H9c2	Cytotoxicity assay		24 hrs	Cytotoxicity against rat H9c2 cells assessed as reduction in cell viability after 24 hrs by MTT assay, IC50=5.2μM	28994286
A549/TR	Antiproliferative assay		72 hrs	Antiproliferative activity against human A549/TR cells after 72 hrs by MTT assay, IC50=1.703μM	29501947
A549	Antiproliferative assay		72 hrs	Antiproliferative activity against human A549 cells after 72 hrs by MTT assay, IC50=2.074μM	29501947
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells after 72 hrs by SRB assay, IC50=0.00025μM	30429953
HT-29	Antiproliferative assay		72 hrs	Antiproliferative activity against human HT-29 cells after 72 hrs by SRB assay, IC50=0.28μM	30429953
HCT8	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT8 cells after 72 hrs by SRB assay, IC50=0.29μM	30429953
HCT116	Function assay		8 hrs	Inhibition of Bmi1 protein expression in human HCT116 cells after 8 hrs by Western blot analysis	30429953
BEAS2B	Function assay		48 hrs	Activation of Keap1/Cul3/Nrf2 in human BEAS2B cells assessed as increase in NQO1 levels measured after 48 hrs, EC50=0.00871μM	30626555
HepG2	Cytotoxicity assay		48 hrs	Cytotoxicity against human HepG2 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay, IC50=0.26μM	31051401
MCF7	Cytotoxicity assay		48 hrs	Cytotoxicity against human MCF7 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay, IC50=0.35μM	31051401
A549	Cytotoxicity assay		48 hrs	Cytotoxicity against human A549 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay, IC50=0.36μM	31051401
A549	Antiproliferative assay		48 hrs	Antiproliferative activity against human A549 cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.52μM	31725288
HepG2	Antiproliferative assay		48 hrs	Antiproliferative activity against human HepG2 cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.52μM	31725288
HOS	Antiproliferative assay		48 hrs	Antiproliferative activity against human HOS cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.66μM	31725288
MCF7	Antiproliferative assay		48 hrs	Antiproliferative activity against human MCF7 cells assessed as inhibition of cell growth incubated for 48 hrs by MTT assay, IC50=0.85μM	31725288
HEK293FT	Function assay		24 hrs	Inhibition of mouse GOAT expressed in HEK293FT cells co-expressing pre-proghrelin assessed as reduction in ghrelin octanoylation incubated for 24 hrs by ELISA, IC50=0.035μM	ChEMBL
MCF-7	Function assay			Inhibitory concentration against proliferation of MCF-7 (ER Positive) breast cancer cells, IC50=0.05μM	15369396
RAW264.7	Anti-inflammatory assay			Anti-inflammatory activity in mouse RAW264.7 cells assessed as inhibition of nitric oxide production, IC50=4μM	28754470
点击查看更多细胞系数据

生物活性

产品描述

特性

Bardoxolone Methyl是一种口服有效的抗炎症调节剂，唯一用于临床实体瘤，2型糖尿病和慢性肾脏疾病的IKKβ抑制剂。

靶点

IKK ^[3]
(Cell-free assay)

Ferroptosis ^[7]

Nrf2 ^[6]

NF-κB ^[6]

体外研究(In Vitro)
体外研究活性	Bardoxolone Methyl作用于小鼠巨噬细胞，对interferon-Ƴ诱导的一氧化氮的产生具有强效的抑制活性，其IC50 为0.1 nM 。^[1] Bardoxolone Methyl降低白血病HL-60，KG-1和NB4细胞的生存能力，其IC50分别为 0.4，0.4，和0.27μM。 CDDO-ME诱导促凋亡Bax蛋白表达，抑制ERK1 / 2的活化，并且它抑制Bcl-2的磷酸化，这有助于诱导细胞凋亡。^[2] Bardoxolone Methyl可有效地抑制(IL)-1beta, phorbol ester, okadaic acid, hydrogen peroxide, lipopolysaccharide, 和cigarette smoke激活的组成型和可诱导的NF-κB的肿瘤坏死因子。^[3]
激酶实验	IKK 试验
激酶实验	对IKK进行分析，以确定CDDO-ME对TNF诱导的IKK活化的效果。简言之，从全细胞提取物的IKK复合物与针对IKKα和IKKβ抗体沉淀，然后用蛋白A / G琼脂糖珠进行处理。 2小时后，用裂解缓冲液清洗珠粒，然后悬浮在含有50 mmol/L HEPES (pH 7.4) 的激酶测定混合物中，20 mmol/L MgCl₂，2 mmol / L的DTT，2 mmol/L DTT, 20 μCi [γ-³²P]ATP，10 μmol/L未标记的ATP和2 μg谷胱甘肽S-转移酶- IκBα（氨基酸1-54）的底物。在30℃下进行温育30分钟，然后加入SDS缓冲液，沸浴5分钟终止该反应。最后，该蛋白在10％SDS-PAGE凝胶中分离，干燥，用Storm820观察记录放射性条带。为了确定每个样品中的IKK-α和IKK-β的总量，将50 μg的全细胞蛋白在7.5％的SDS-PAGE下解析，电子转移至硝酸纤维素膜上，然后与抗-IKK-α或抗-IKK-β的抗体印迹杂交。
细胞实验	细胞系	HL-60, KG-1, 和 NB4 细胞
	浓度	~5 μM
	孵育时间	72 小时
	方法	白血病细胞系以3.0 × 10⁵ cells/mL的密度进行培养，同时白血病单核细胞以5 × 10⁵ cells/mL的密度置于存在或不存在显示浓度的CDDO-ME中培养。加入适量的 DMSO（终浓度小于0.05％）作为对照。添加1 μM ara-C到培养基，用以细胞毒性研究。 24至72小时后，用血细胞计数板的台盼蓝染料排除法进行存活细胞计数。
实验图片	检测方法	检测指标	实验图片	PMID
	Western blot	p-IκBα / IκBα Bcl-xl / Bcl-2 / Bax / Cleaved caspase / Cytochrome C / PARP / Cleaved PARP p-PI3K / PI3K / p-AMPK / AMPK / p-p38 MAPK / p38 MAPK / p-AKT / AKT / p-mTOR / mTOR PTEN / PP2A / PHLPP1		25897966
	Immunofluorescence	PDI / SDHA c-PARP / Cytochrome C / COX IV		26053096
	Growth inhibition assay	Cell viability		25733817

体内研究(In Vivo)
体内研究活性	Bardoxolone Methyl(60 mg/kg)体内用药，可减少肺肿瘤的数量，大小和降低严重程度。 ^[4] Bardoxolone Methyl还可显著降低LPS刺激下的体内炎症因子的反应，诱导脾脏HO-1蛋白表达，对抗致死剂量的LPS保护小鼠。 ^[5]
动物实验	Animal Models	雌性 A/J 小鼠腹腔注射 Vinyl carbamate.
	Dosages	~60 mg/kg
	Administration	口服给药

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
临床试验信息 (data from https://clinicaltrials.gov, updated on 2024-01-11)
NCT02316821	Completed	Chronic Kidney Disease\|Type 2 Diabetes	Kyowa Kirin Co. Ltd.	December 2014	Phase 2
NCT02036970	Completed	Pulmonary Arterial Hypertension\|Pulmonary Hypertension\|Interstitial Lung Disease\|Idiopathic Interstitial Pneumonia\|Idiopathic Pulmonary Fibrosis\|Sarcoidosis\|Respiratory Bronchiolitis Associated Interstitial Lung Disease\|Desquamative Interstitial Pneumonia\|Cryptogenic Organizing Pneumonia\|Acute Interstitial Pneumonitis\|Idiopathic Lymphoid Interstitial Pneumonia\|Idiopathic Pleuroparenchymal Fibroelastosis	Biogen	May 31 2014	Phase 2
NCT01598363	Completed	Healthy Volunteers	Biogen	June 30 2012	Phase 1
NCT01551446	Withdrawn	Renal Insufficiency Chronic\|Diabetes Mellitus Type 2	Biogen	April 30 2012	Phase 1
NCT01503866	Completed	Healthy	Biogen	December 1 2011	Phase 1

参考文献
[1] Honda T, et al. J Med Chem. 2000, 43(22), 4233-4246. [2] Konopleva M, et al. Blood. 2002, 99(1), 326-335. [3] Shishodia S, et al. Clin Cancer Res. 2006, 12(6), 1828-1838. [4] Liby K, et al. Cancer Res. 2007, 67(6), 2414-2419. [5] Auletta JJ, et al. J Interferon Cytokine Res. 2010, 30(7), 497-508. [6] Tanjoni I, et al. Cancer Biol Ther. 2010, 15;9(10):764-77. [7] Chengcheng Gai, et al. J Cell Physiol . 2020 Apr;235(4):3329-3339. + 展开

参考文献

+ 展开

化学信息&溶解度

分子量	505.69	分子式	C₃₂H₄₃NO₄
CAS号	218600-53-4	SDF	Download Bardoxolone Methyl SDF
Smiles	CC1(CCC2(CCC3(C(C2C1)C(=O)C=C4C3(CCC5C4(C=C(C(=O)C5(C)C)C#N)C)C)C)C(=O)OC)C
储存条件(自收到货起)	3年-20°C粉状	储备液保存和期限建议分装储备液，避免反复冻融！	1年-80°C溶于溶剂
储存条件(自收到货起)	3年-20°C粉状	储备液保存和期限建议分装储备液，避免反复冻融！	1个月-20°C溶于溶剂

体外溶解度
批次：

DMSO : 26 mg/mL ( (51.41 mM)； DMSO吸湿会降低化合物溶解度，请使用新开封DMSO)

Water : Insoluble

Ethanol : Insoluble

摩尔浓度计算器

体内溶解度
批次：

现配现用，请按从左到右的顺序依次添加，澄清后再加入下一溶剂

动物体内配方计算器

实验计算

摩尔浓度计算器

质量	浓度	体积	分子量
=	×	×

稀释计算器分子量计算器

动物体内配方计算器（澄清溶液）

第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量）

给药剂量 mg/kg 动物平均体重 g 每只动物给药体积 μL 动物数量只

第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系Selleck为您提供正确的澄清溶液配方）

% DMSO + % + % Tween 80 + % ddH2O

%DMSO+ %

计算结果：

工作液浓度： mg/ml；

DMSO母液配制方法： mg 药物溶于μL DMSO溶液（母液浓度mg/mL，注：如该浓度超过该批次药物DMSO溶解度，请先联系Selleck）；

体内配方配制方法：取μL DMSO母液，加入μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入μL ddH₂O，混匀澄清。

体内配方配制方法：取μL DMSO母液，加入μL Corn oil，混匀澄清。

注意：1. 首先保证母液是澄清的；
2.一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。

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C1=C0/X	C1:	LOG(C1):
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C5=C4/X	C5:	LOG(C5):
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