Difenoconazole   中文名称: 苯醚甲环唑

苯醚甲环唑对禾谷镰刀菌有抑制作用，EC50为1.69～19.6 mg/L[2]。苯醚甲环唑的 EC50 值分别为 0.131 和 0.131，可抑制立斑病、黄腐病、灰葡萄孢和立枯丝核菌的发育。 0.297、0.252 和 0.069 毫克/升[2]。

苯醚甲环唑（0.25-2 mg/L；暴露 96 小时）可诱导斑马鱼细胞生长表现出一系列症状，包括生长抑制、心脏减慢、生长衰退和形态发生 [1]。

Animal/Disease Models: Zebrafish [1]
Doses: 0.25, 0.5, 1, 1.5 and 2 mg/L
Route of Administration: 96 hrs (hrs (hours)) of exposure
Experimental Results: Within 24 hrs (hrs (hours)) at a concentration of 0.5 mg/L, the body color of zebrafish larvae changed Dramatically Black, heart rate decreases. Inhibition of growth weight was measured in adult zebrafish after 14 days of exposure to 0.25 mg/L.

Absorption, Distribution and Excretion
Following a single oral administration of 0.5 mg/kg body weight of [(14)C-phenyl]fenofibrate, both male and female rats experienced rapid and almost complete absorption. A single oral administration of 300 mg/kg body weight of [(14)C-phenyl]fenofibrate resulted in lower absorption, with 17% and 22% of the dose excreted in feces in cannulated male and female rats, respectively. Peak plasma concentrations were reached in the low-dose group within 2 hours, followed by a rapid decline; peak plasma concentrations were reached in the high-dose group approximately 4 hours later, followed by a slow decline over 24 hours, with a similar rate of decline to the low-dose group. The difference in area under the plasma concentration-time curve (AUC) between the low-dose and high-dose groups in female rats corresponded to the dose difference, while the AUC difference in male rats was 400-fold. The systemic dose was primarily excreted via bile, with 73% of the administered dose (0.5 mg/kg body weight) excreted in bile in male rats and 76% in female rats. In cannulated rats, 14% of the administered dose was excreted in urine, 9% in female rats, and less than 4% in feces, confirming that the drug was almost completely absorbed at low doses. When bile (0.5 mg/kg body weight) from male rats was administered duodenally to other cannulated rats, 80% of the dose was re-excreted in bile, with only 4% excreted in urine, indicating enterohepatic circulation. At a dose of 300 mg/kg body weight, bile was also the main route of excretion, accounting for 56% of the administered dose in male rats and 39% in female rats, while urinary excretion accounted for only 1%. In uncannulated male and female rats, after a single oral administration of 0.5 mg/kg body weight of [(14)C-phenyl]fenofibrate or [(14)C-triazole]fenofibrate, 13-22% of the administered dose was excreted in urine and 81-87% in feces. In uncannulated male and female rats, following a single oral administration of 300 mg/kg body weight of [(14)C-phenyl]fenofibrate or [(14)C-triazole]fenofibrate, 8–15% of the administered dose was excreted in the urine and 85–95% in the feces. At any dose, there were no significant differences in excretion profiles between male and female rats or between the two radiolabeled forms. When rats were given similar doses of [(14)C-phenyl]fenofibrate or [(14)C-triazole]fenofibrate after 14 consecutive days of oral administration of 0.5 mg/kg body weight of unlabeled fenofibrate, no sex differences were observed, and the excretion profiles were not significantly different from those of untreated rats. Excretion data from the 0.5 mg/kg body weight dose group showed that although enterohepatic circulation was present, bile metabolites were primarily excreted in the feces. At lower doses, the excretion half-life of the drug was approximately 20 hours. At a dose of 300 mg/kg body weight, uncannulated rats excreted more of the administered dose in their urine than cannulated rats, presumably due to the reabsorption and further metabolism of some bile metabolites; however, as observed in the low-dose group, the primary route of excretion of radioactive material in bile remained feces. In the high-dose group, the excretion half-life was approximately 33–48 hours. Therefore, at both doses, elimination kinetics were independent of sex and the location of the radiolabel. In male rats, after oral administration of a 0.5 mg/kg body weight dose, the concentration of the radiolabeled material in the blood reached its peak concentration (Cmax) at 2 hours, followed by a rapid decline. The AUC within 168 hours post-administration was 6.19 μg equivalents/hour/mL. The Tmax in female rats was shorter than in male rats, and the Cmax and AUC values in female rats were only about 50% of the corresponding values in male rats. The rate of elimination of radioactive material from the body was slightly faster in female rats than in male rats. Tissue clearance results showed that, at 2 and 24 hours after administration of 0.5 mg/kg body weight of [(14)C-phenyl]fenofibrate, only the liver and kidneys showed persistently higher radioactive concentrations than plasma concentrations in both male and female rats. Similar results were observed in whole-body autoradiographic sections of male rats administered the same dose, as most of the radioactive material was present in the gastrointestinal contents and bile at 2 and 24 hours after administration, with lower concentrations in the liver and kidneys. Other tissues with initially higher radioactive concentrations than plasma included the adrenal glands of both male and female rats, and the Haversian glands and adipose tissue of female rats; however, the radioactive concentrations in these tissues decreased rapidly. After 168 hours, the residual levels of [(14)C-phenyl]fenofibrate in tissues were extremely low, with only the concentration in adipose tissue comparable to that in plasma, while the residual levels of [(14)C-triazole]fenofibrate in all tissues were below the limits of detection or quantitation. For both radiolabeled difenoconazoles, residual levels in female animal tissues were generally slightly lower than in male animals, and pretreatment with the unlabeled test substance had no effect on tissue distribution. Tissue clearance results showed that 4 hours after administration of [(14)C-phenyl]difenoconazole at a dose of 300 mg/kg body weight, concentrations in most tissues of both sexes were similar to or higher than plasma concentrations. The highest concentrations were found in adipose tissue, followed by the liver, Haschs gland, adrenal gland, kidney, and pancreas. In all other tissues, drug concentrations initially higher than plasma concentrations decreased rapidly within 48 hours after administration, and by 168 hours, residual levels of [(14)C-phenyl]difenoconazole in all tissues were significantly lower, except for adipose tissue where residual levels were higher than plasma concentrations. Tissue residual levels of [(14)C-triazole]difenoconazole were significantly lower than those of [(14)C-phenyl]difenoconazole, and by 168 hours, it was only detectable in the liver. Measurements of gastrointestinal contents were consistent with the observed absorption and elimination curves. Following 14 consecutive days of administration of [4-chlorophenoxy-U-(14)C]fenofibrate at a dose of 0.5 mg/kg body weight/day, the absorbed radiolabeled material was rapidly and almost completely eliminated, primarily via feces. Over 90% of the radiolabeled material was eliminated within 24 hours of the last dose, and 98.5% was recovered within 7 days. At this point, less than 0.5% of the radiolabeled material remained in tissues and the carcass. Although some quantitative differences were observed between single and multiple oral administrations, the metabolite profiles in urine and feces were qualitatively similar at various time points. Radiolabeled material concentrations plateaued in most tissues after 7 days. In the liver, kidneys, fat, and pancreas, residual concentrations increased with continued administration and did not plateau during the administration period; however, it was estimated that residual concentrations would plateau within 3 weeks. Clearance of radiolabeled material from tissues was moderately rapid. Assuming that the clearance of radiolabeled compounds from tissues follows first-order kinetics and single-phase consumption kinetics, with a half-life typically of 4–6 days, clearance is faster in the liver, kidneys, and pancreas, with a half-life of 1–3 days; clearance is slower in adipose tissue, with a half-life of 9 days. Experiments using position-specific radiolabeled compounds showed that triazole-labeled compounds had the highest tissue concentrations in the liver, while phenyl-labeled compounds had the highest tissue concentrations in adipose tissue and plasma. The residual amounts of triazole-labeled compounds were significantly lower than those of phenyl-labeled compounds. The tissue residual amounts in women were slightly lower than those in men. Repeated pretreatment with unlabeled difenoconazole had no effect on tissue distribution.
This study investigated the in vivo skin penetration of unlabeled difenoconazole (99.3% purity) and [triazole-U-(14)C]difenoconazole after dermal administration to rats, as well as in vitro studies on rat and human skin. In male HanBrlWIST (SPF) rats, the absorption, distribution, and excretion of radioactive materials were investigated after a single dermal administration of a mixture of (14)C-labeled difenoconazole and unlabeled difenoconazole (prepared as SCORE 250 EC). The specific activity of the highest dose group was 54 kBq/mg (1.5 μCi/mg). The difenoconazole in the highest dose group was dissolved in a blank preparation at a concentration of 250 g/L, representing the undiluted product. For the intermediate and lowest doses, the prepared materials were mixed with water at ratios of 1:200 (w/v) and 1:5000 (w/v), respectively. The nominal doses administered were 0.5, 13, and 2600 μg/cm², respectively, and remained for 6 hours. Dermal absorption at the highest dose was repeated in another experiment at an administration dose of 2400 μg/cm². …At the lowest, intermediate, and highest doses, the systemic absorption rates over 6 hours were 15.3%, 7.5%, and 7.1%, respectively, with permeability rates of 0.013, 0.162, and 30.4 μg/cm²/hour, respectively. These permeability ratios (i.e., 1:12:2400) were proportional to the administered concentration (i.e., 1:26:5100). However, significant variability exists in the data, attributed to the irritant nature of the formulation carrier SCORE 250 EC; this resulted in a wide range of individual values for skin absorption, reaching up to 45% of the administered dose. In in vivo rat studies, the mean skin absorption rates at the lowest, intermediate, and highest doses were 37.6%, 14.6%, and 10.6% in the worst-case scenario. Nevertheless, residual drug concentrations in the blood were generally very low: the highest concentrations (in difenoconazole equivalents) at the intermediate and highest doses were 0.01 μg/mL and 0.26 μg/mL, respectively, 6–8 hours after administration. In vitro, after exfoliation of the stratum corneum from rat and human epidermal membranes, the permeability of non-radiolabeled difenoconazole (99.3% purity) and [triazole-U-14C] difenoconazole to the separated epidermal membranes was determined after 24 hours of exposure in difenoconazole solutions at concentrations of 0.05, 1.28, or 250 mg/mL. The doses of difenoconazole administered were 0.5, 12, or 2345 μg/cm². …Over 24 hours, the percentages of radiolabeled substances penetrating the epidermal membranes at the lowest, intermediate, and highest concentrations were: 71%, 64%, and 23% for rat skin epidermis; and 7.6%, 7.0%, and 0.7% for human skin epidermis. Although in vitro human skin studies showed that the skin absorption rate at the lowest concentration was approximately 8% (7.6%), this could be increased to 15% if the residue in the skin is considered a potentially absorbable substance. However, the primary objective of the in vitro rat and human skin comparison study was to assess the differences in actual percentage absorption, thereby estimating the appropriate species ratio. At the lowest, intermediate, and highest concentrations, the flux (i.e., the rate of permeation under steady-state conditions) of rat skin was 0.020, 0.455, and 26.0 μg/cm²/hr, respectively, while that of human skin was 0.002, 0.037, and 0.82 μg/cm²/hr, respectively. Therefore, at the lowest, intermediate, and highest concentrations, the flux ratios of rat to human were approximately 1:10, 1:12, and 1:32, respectively. Comparing permeability at different doses (concentration ratio of 1:25:5000), the permeability of the rat epidermal membrane increased to 1:23:1300, while the permeability of the human epidermal membrane was only 1:24:500. Metabolites/Metabolites were isolated from the urine and feces of male and female rats. These rats were administered a single oral dose of 0.5 or 300 mg/kg body weight of [(14)C-phenyl]fenofibrate or [(14)C-triazole]fenofibrate, or a pre-administered oral dose of 0.5 mg/kg body weight of unlabeled fenofibrate 14 times daily, followed by an oral dose of 0.5 mg/kg body weight of fenofibrate. Balance data for phenyl and triazole ring labels showed that over 97% of the radiolabeled material was excreted in all cases, with over 78% excreted in feces. Three major metabolites, A, B, and C, were isolated from the feces, accounting for an average of 68% of the administered dose. Metabolite B (hydroxy-CGA 169374) was hydroxylated on the outside of the benzene ring, and spectral analysis revealed that it contained two isomers, one of which had a rearrangement of the chlorine atom on the outside of the benzene ring, attributed to a mechanism similar to the NIH shift. Metabolite C (CGA 205375) is the hydroxyl product of the dioxolane cleavage in the dioxolane molecule and is found only in the feces of rats receiving higher doses of the drug. Metabolite A (hydroxy-CGA 205375) is the hydroxylated product of the benzene ring of metabolite C, containing two diastereomers, similar to metabolite B. The chromatograms of urinary metabolites are more complex, with greater differences between the two radiolabeled forms. Free 1,2,4-triazole was detected in male urine at levels below 10% of the administered dose, representing the breaking of intercyclic alkyl bridges. Other urinary metabolites include metabolite C and its sulfate conjugate, cyclically hydroxylated metabolite C and its sulfate conjugate, and the chlorophenoxy-chlorophenyl moiety of glycolic acid metabolites, all at low levels, each below 3% of the administered dose. In addition, a metabolite CGA 189138 (chlorophenoxy-chlorobenzoic acid) was isolated from the liver. Therefore, diphenyl ether dicyclazole is extensively metabolized, although the cleavage of the triazole ring and dioxolane ring is limited. The observed extensive bile excretion is consistent with the relatively high molecular weight of the major metabolite. The major steps in the inferred metabolism of diphenoconazole in rats include the hydrolysis of the ketal in diphenoconazole to generate CGA 205375 (1-[2-chloro-4-(4-chlorophenoxy)phenyl]-2-(1,2,4-triazole)-1-ylethanol), in which the ketone CGA 205374 (1-(2-chloro-4-(4-chlorophenoxy)phenyl)-2-(1,2,4-triazole)-1-ylethyl ketone) is considered a speculative but unidentified intermediate, and hydroxylation of the outer benzene ring of the parent compound (hydroxy-CGA 169374) and CGA 205375 (hydroxy-CGA 205375). As a secondary process, the alkyl chain between the triazole ring and the inner benzene ring breaks, generating glycolic acid (NOA 448731) or CGA 189138 (2-chloro-4-(4-chlorophenoxy)benzoic acid) and free triazole. Sulfate conjugates of CGA 205375 and hydroxy-CGA 205375 have been identified. The dissipation of the fungicide difenoconazole (3-chloro-4-[(2RS,4RS;2RS,4SR)-4-methyl-2-(1H-1,2,4-triazol-1-ylmethyl)-1,3-dioxolane-2-yl]phenyl-4-chlorophenyl ether) after application to apples used in the production of infant food was investigated. To control pathogens causing fungal diseases, this formulation was sprayed on three apple varieties (Jonagold Decosta, Gala, and Idared) containing powdery mildew (Podosphaera leucotricha ELL et Ev./Salm.) and apple scab (Venturia inaequalis Cooke/Aderh.). Residue analysis was performed using a validated gas chromatography-electron capture detector (GC-ECD) and nitrogen-phosphorus detector (NPD) method. The analytical performance of this method was very satisfactory, with an expanded uncertainty of 19% (coverage factor k = 2, confidence level 95%). The dissipation of difenoconazole was studied using a pseudo-first-order kinetic model (determination coefficient R² between 0.880 and 0.977). In experiments conducted on the three apple varieties, the half-life of difenoconazole ranged from 12 to 21 days. In these experiments, the initial residue levels gradually decreased, reaching 0.01 mg/kg within 50–79 days. To maintain residue levels below 0.01 mg/kg (the maximum acceptable concentration for infant formula), difenoconazole must be applied approximately 3 months prior to harvest at a dose of 0.2 L/ha (50 g of active ingredient per hectare).

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Biological half-life
After 14 days of application of [4-chlorophenoxy-U-(14)C]difenoconazole at a dose of 0.5 mg/kg body weight/day, the clearance rate of radioactive residues in tissues was moderate. Assuming that the clearance of radiolabeled substances from tissues follows first-order kinetics and single-phase clearance kinetics, the half-life is typically 4–6 days. Clearance is faster in the liver, kidneys, and pancreas, with half-lives of 1–3 days, while clearance is slower in fat, with a half-life of 9 days.

Toxicity Summary
Identification and Uses: Difenoconazole is a white crystalline solid. It can be used as a fungicide, insecticide, and seed treatment/protectant. Human Exposure and Toxicity: Harmful if inhaled or absorbed through the skin. May cause moderate eye irritation. Has not caused chromosomal aberrations in human lymphocytes. One case of allergic reaction to the formulation has been reported. Animal Studies: Difenoconazole has moderate transient eye irritation in rabbits. Has mild transient skin irritation in rabbits. When applied topically to intact rabbit skin, it is considered essentially non-toxic. In rats, after 28 days of transdermal administration, the incidence of mild central lobular hepatocyte hypertrophy was increased in both male and female rats at a dose of 1000 mg/kg body weight. In the thyroid gland, the incidence of mild to moderate follicular epithelial hypertrophy was slightly increased in the 1000 mg/kg body weight dose group. The liver appears to be a target organ for toxicity. No evidence of carcinogenicity or tumorigenicity in rats has been found. In rabbits, no embryotoxicity, fetal toxicity, or teratogenicity was observed at doses up to 75 mg/kg body weight/day. In rats, no embryotoxicity or teratogenicity was observed at doses up to 200 mg/kg body weight. Microscopic examination of the central and peripheral nervous systems of rats showed that neither male nor female rats were affected at dietary concentrations of difenoconazole up to 1500 ppm. Difenoconazole did not induce gene mutations in bacterial cells or cultured mammalian cells. Ecotoxicity studies: In zebrafish experiments, different doses of difenoconazole induced a range of symptoms in embryonic development, including hatching inhibition, abnormal spontaneous movement, slowed heart rate, growth regression, and morphological deformities. Difenoconazole exposure altered thyroid hormone levels and gene transcription in zebrafish larvae, indicating endocrine disruption. Difenoconazole upregulated gene expression in zebrafish embryos associated with hatching, retinoic acid metabolism, and lipid homeostasis. Exposure to difenoconazole also altered lipid metabolism and lipid profiles in marine medaka (Oryzias melastigma). Difenoconazole inhibited mitochondrial respiration in the flight muscle of bumblebees.

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Non-human toxicity values
Duck oral LD50 >2150 mg/kg
Rabbit dermal LD50 >2010 mg/kg
Rat inhalation LC50 >45 mg/cu m/4 hr
Rat oral LD50 1453 mg/kg
For more complete non-human toxicity data for difenoconazole (7 items), please visit the HSDB record page.

[1]. Evaluation of acute and developmental effects of difenoconazole via multiple stage zebrafish assays. Environ Pollut. 2013 Apr;175:147-57.

[2]. Chiral triazole fungicide difenoconazole: absolute stereochemistry, stereoselective bioactivity, aquatic toxicity, and environmental behavior in vegetables and soil. Environ Sci Technol. 2013 Apr 2;47(7):3386-94.

Difenoconazole belongs to the dioxolane class of compounds, with a structure of 1,3-dioxolane ring, substituted at position 2 with 2-chloro-4-(4-chlorophenoxy)phenyl and 1,2,4-triazol-1-ylmethyl. It is a broad-spectrum fungicide with novel broad-spectrum activity, usable as a spray or seed treatment. It exhibits moderate toxicity to humans, mammals, birds, and most aquatic organisms. It is an environmental pollutant, exogenous substance, EC 1.14.13.70 (sterol 14α-demethylase) inhibitor, and antifungal pesticide. It is an aromatic ether, dioxolane, triazole compound, cyclic ketal, azole fungicide, and triazole fungicide. Difenoconazole is a broad-spectrum fungicide effective against a variety of fungi, including members of the classes Asporidium, Basidiomycetes, and Deuteromycetes. It can be used as a seed treatment, foliar spray, and systemic fungicide. It is absorbed through the surface of infected plants and transported to various parts of the plant. It has both therapeutic and preventative effects. Difenoconazole can be used on winter wheat, rapeseed, Brussels sprouts, cabbage, broccoli/Calabras broccoli, and cauliflower. It controls a variety of fungi, including wheat glume blight, brown rust, pale leaf spot, leaf spot, pod spot, ring spot, and stem rot. It can also prevent discoloration of winter wheat ears. The mechanism of action of difenoconazole is to inhibit sterol demethylation, thereby preventing the biosynthesis of ergosterol in the fungal cell membrane, and thus inhibiting fungal growth and development.

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Mechanism of Action
Difenoconazole can be applied through foliar spraying or seed treatment. Its mechanism of action is to inhibit the 14α-demethylation of sterols, interfering with the synthesis of ergosterol in the target fungi, thereby causing changes in the morphology and function of the fungal cell membrane.

C19H17CL2N3O3

406.26

405.064

119446-68-3

86173

White to off-white solid powder

1.4±0.1 g/cm3

547.0±60.0 °C at 760 mmHg

76°C

284.6±32.9 °C

0.0±1.5 mmHg at 25°C

1.642

4.92

58.4

0

5

5

27

495

0

BQYJATMQXGBDHF-UHFFFAOYSA-N

InChI=1S/C19H17Cl2N3O3/c1-13-9-25-19(27-13,10-24-12-22-11-23-24)17-7-6-16(8-18(17)21)26-15-4-2-14(20)3-5-15/h2-8,11-13H,9-10H2,1H3

1-((2-(2-chloro-4-(4-chlorophenoxy)phenyl)-4-methyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazole

Plover ScoreBardos Neu CGA-169374 CGA169374CGA 169374 DragonDifenoconazole

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~246.15 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.15 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (6.15 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (6.15 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。