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Infigratinib (also known as BGJ398; BGJ-398; NVP-BGJ398; Truseltiq) is a novel, potent, selective, and orally bioavailable FGFR (fibroblast growth factor receptors) inhibitor with potential antiangiogenic and antineoplastic activities. In order to treat cholangiocarcinoma, the FDA has approved it as an anti-cancer medication as of May 28, 2021. With an IC50 of 0.9 nM/1.4 nM/1 nM in cell-free assays, infigratinib inhibits FGFR1/2/3. FGFR is the target of over 40-fold selectivity when compared to FGFR4, VEGFR2, and Abl, Fyn, Kit, Lck, Lyn, and Yes are the targets of minimal activity. An orthotopic xenograft bladder cancer model in mice demonstrates strong anti-proliferative activity in vitro and strong in vivo antitumor efficacy.
Physicochemical Properties
| Molecular Formula | C26H31CL2N7O3 |
| Molecular Weight | 560.48 |
| Exact Mass | 559.186 |
| Elemental Analysis | C, 55.72; H, 5.57; Cl, 12.65; N, 17.49; O, 8.56 |
| CAS # | 872511-34-7 |
| Related CAS # | Infigratinib phosphate;1310746-10-1 |
| PubChem CID | 53235510 |
| Appearance | White to off-white solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 747.9±60.0 °C at 760 mmHg |
| Flash Point | 406.1±32.9 °C |
| Vapour Pressure | 0.0±2.5 mmHg at 25°C |
| Index of Refraction | 1.654 |
| LogP | 5.03 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 38 |
| Complexity | 724 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O=C(N(C)C1C=C(NC2C=CC(N3CCN(CC)CC3)=CC=2)N=CN=1)NC1C(Cl)=C(OC)C=C(OC)C=1Cl |
| InChi Key | QADPYRIHXKWUSV-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C26H31Cl2N7O3/c1-5-34-10-12-35(13-11-34)18-8-6-17(7-9-18)31-21-15-22(30-16-29-21)33(2)26(36)32-25-23(27)19(37-3)14-20(38-4)24(25)28/h6-9,14-16H,5,10-13H2,1-4H3,(H,32,36)(H,29,30,31) |
| Chemical Name | 3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-[6-[4-(4-ethylpiperazin-1-yl)anilino]pyrimidin-4-yl]-1-methylurea |
| Synonyms | NVP-BGJ398; Infigratinib; NVPBGJ398; BGJ398; NVP-BGJ398; BGJ-398; BGJ398; BGJ 398; 3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1-methylurea; Infigratinib [INN]; BGJ-398; NVPBGJ 398; NVPBGJ-398; BG J398 |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
FGFR1 (IC50 = 0.9 nM); FGFR2 (IC50 = 1.4 nM); FGFR3 (IC50 = 1 nM); FGFR4 (IC50 = 60 nM) Fibroblast Growth Factor Receptor (FGFR) 1 (IC50 = 0.9 nM), FGFR2 (IC50 = 1.4 nM), FGFR3 (IC50 = 1.0 nM), FGFR4 (IC50 = 37 nM); weak activity against VEGFR2 (IC50 = 65 nM), PDGFRα (IC50 = 80 nM); no activity against EGFR, ALK (IC50 > 1000 nM) [1] - FGFR1/2/3 (validated in FGFR-amplified endometrial cancer cells; no additional IC50 values) [2] - FGFR (targeted in FGFR-signaling activated triple-negative breast cancer (TNBC); consistent with [1]’s IC50 data for FGFR1-3) [3] |
| ln Vitro |
BGJ398 also inhibits VEGFR2 to a lesser extent. BGJ398 has an IC50 of 0.18 μM for inhibiting VEGFR2. Other kinases such as ABL, FYN, KIT, LCK, LYN, and YES are suppressed by BGJ398 with IC50 values of 2.3 μM, 1.9 μM, 0.75 μM, 2.5 μM, 0.3 μM, and 1.1 μM, in that order. BGJ398 impedes the growth of BaF3 cells that are dependent on FGFR1, FGFR2-Q, and FGFR3 at the cellular level, with IC50 values of 2.9 μM, 2.0 μM, and 2 μM, in that order. BGJ398 has an IC50 of 4.6 nM, 4.9 nM, 5 nM, 5 nM, and 168 nM, respectively, and inhibits autophosphorylation on particular tyrosine residues, such as FGFR-WT, FGFR2-WT, FGFR3-K650E, FGFR3-S249C, and FGFR4-WT. Cancer cells that overexpress wild-type (WT) FGFR3, such as RT112, RT4, SW780, and JMSU1, are suppressed by BGJ398 with IC50 values of 5 nM, 30 nM, 32 nM, and 15 nM, respectively.[1] Inhibited recombinant FGFR kinase activity: FGFR1 (IC50 = 0.9 nM), FGFR2 (IC50 = 1.4 nM), FGFR3 (IC50 = 1.0 nM); suppressed FGFR autophosphorylation in Ba/F3-FGFR1 cells (IC50 = 3.3 nM), Ba/F3-FGFR2 cells (IC50 = 4.2 nM) [1] - Reduced viability of FGFR-amplified endometrial cancer cells: AN3CA (IC50 = 12 nM), HEC-1A (IC50 = 25 nM), Ishikawa (IC50 = 48 nM); 100 nM Infigratinib downregulated p-ERK1/2 (Thr202/Tyr204) and p-AKT (Ser473) in AN3CA cells (2 hours) [2] - Inhibited proliferation of FGFR-driven TNBC cells: MDA-MB-453 (IC50 = 18 nM), BT-549 (IC50 = 22 nM); blocked FGFR-mediated p-STAT3 (Tyr705) phosphorylation in MDA-MB-453 cells (50 nM, 4 hours) [3] |
| ln Vivo |
BGJ398 inhibits tumor growth and induces stasis in this orthotopic xenograft bladder cancer model after being taken orally for 12 days straight at doses of 10 and 30 mg/kg, respectively. It's interesting to note that the animals that received BGJ398 show either 10% body weight gain (30 mg/kg) or no body weight loss (10 mg/kg), which is another sign of efficacy. One oral dose of BGJ398 monophosphate salt at 4.25 and 8.51 mg/kg is given to female Rowett rats (RT112) that are tumor-bearing. In a dose-dependent manner, BGJ398 dramatically lowers the levels of pFRS2 and pMAPK. In a dose-dependent manner, BGJ398 markedly suppresses bFGF-stimulated angiogenesis. Nevertheless, BGJ398 does not impede the formation of blood vessels induced by VEGF. In nude mice bearing H1581 (FGFR1-amplified lung cancer) xenografts: Oral Infigratinib (15 mg/kg/day) for 21 days resulted in 85% tumor growth inhibition (TGI); tumor p-FGFR1 reduced by 70% (immunoblotting) [1] - In nude mice bearing AN3CA (FGFR2-amplified endometrial cancer) xenografts: Oral Infigratinib (20 mg/kg/day) for 14 days achieved 72% TGI; no tumor regression but suppressed p-ERK in tumor tissues [2] - In syngeneic TNBC mouse model (4T1-FGFR3): Oral Infigratinib (25 mg/kg/day) for 28 days caused 68% TGI; improved mouse survival (median survival: 42 days vs. 28 days for vehicle) [3] |
| Enzyme Assay |
The purified GST-fusion FGFR3-K650E kinase domain phosphorylates a synthetic substrate in the presence of radiolabeled ATP to measure the enzymatic kinase activity. The enzyme activity is determined by combining 10 μL of the corresponding substrate mixture (peptidic substrate, ATP, and [γ 33 P]ATP) with 10 μL of a 3-fold concentrated BGJ398 solution or control. The assay buffer is mixed with 10 μL of a concentrated enzyme solution three times over to start the reactions. The following are the assay components' final concentrations: 0.5 μM ATP (γ-[ 33 P]-ATP 0.1 μCi), 3 mM MnCl2, 3 mM MgCl2, 1 mM DTT, 250 μg/mL PEG 20000, 2 μg/mL poly(EY) 4:1, 1% DMSO, and 10 ng of GST-FGFR3-K650E were added. The assay is performed using the filter binding (FB) method in 96-well plates for 10 minutes at room temperature in a final volume of 30 μL with BGJ398 included. When 20 μL of 125 mM EDTA is added, the enzymatic reactions are stopped, and the amount of 33 P incorporated into the polypeptidic substrates is measured as follows: 30 microliters of the halted reaction mixture are placed onto Immobilon-PVDF membranes that have been soaked in methanol for five minutes, rinsed with water, and then soaked in 0.5% H3PO4 for five minutes. The membranes are then mounted on a vacuum manifold that has a disconnected vacuum source. Following spotting, a vacuum is connected, and 200 μL of 0.5% H3PO4 is rinsed through each well. Free membranes are taken out and ished with 1% H3PO4 four times and ethanol once on a shaker. Membranes are dehydrated and covered with a layer of scintillation fluid (10 μL/well). Eventually, a microplate scintillation counter is used to seal and count the plates. The BGJ398 percentage inhibition is analyzed using linear regression to determine the IC50 values. FGFR kinase activity assay: Recombinant human FGFR1/2/3/4 kinases (50 ng/well) were incubated with 10 μM ATP and a fluorescent peptide substrate in reaction buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl2, 1 mM DTT) at 30°C for 60 minutes. Infigratinib (0.1 nM-1 μM) was added 15 minutes before ATP initiation. Phosphorylated substrate was detected via fluorescence polarization; IC50 values were calculated using nonlinear regression (four-parameter logistic model) [1] |
| Cell Assay |
The RPMI-1640 medium supplemented with 10% FBS, 4.5 g/L glucose, 1.5 g/L sodium bicarbonate, and Pen/Strep is used to cultivate mouse BaF3 cell lines, whose proliferation and survival have been rendered IL-3-independent by stable transduction with tyrosine kinases activated either by mutation or fusion with a dimerizing partner. Twice a week, cells are passaged. The Luciferase bioluminescent assay is utilized to evaluate the BGJ398-mediated suppression of BaF3 cell proliferation and viability. BaF3 or BaF3 Tel-TK cells that are growing exponentially are seeded into 384-well plates (4250 cells/well) at a volume of 50 μL per well using a μFill liquid dispenser in fresh medium. In a polypropylene 384-well plate, BGJ398 is serially diluted in DMSO. The plates were then incubated at 37 °C (5% CO2) for 48 hours after 50 nL of BGJ398 were added using the pintool transfer device. The luminescence is then measured with an Analyst-GT after adding 25 μL of Bright-Glo. The logarithm of inhibitor concentration is used to generate a logistic fit of the percent cell viability. This is done using custom curve-fitting software. When cell viability is reduced to 50% of a DMSO control, the concentration of BGJ398 is called the IC50 value. FGFR-engineered cell proliferation assay (Ba/F3-FGFR1/2/3): Cells were seeded in 96-well plates (5×10³ cells/well) and treated with Infigratinib (0.1 nM-1 μM) for 72 hours. Cell viability was measured via tetrazolium salt reduction assay; absorbance at 570 nm was recorded, and IC50 values were determined [1] - Endometrial cancer cell assay (AN3CA/HEC-1A): Cells were seeded at 4×10³ cells/well, treated with Infigratinib (0.1 nM-1 μM) for 72 hours. Viability was assessed via colorimetric assay; Western blot analyzed p-FGFR, p-ERK, p-AKT, and GAPDH (30 μg protein/ lane, 10% SDS-PAGE, chemiluminescence detection) [2] - TNBC cell assay (MDA-MB-453): Cells were seeded at 6×10³ cells/well, treated with Infigratinib (1 nM-500 nM) for 96 hours. Viability was measured via fluorometric ATP assay; caspase-3/7 activity was detected to assess apoptosis (200 nM, 24 hours) [3] |
| Animal Protocol |
Mice: Female HsdNpa: It is done with athymic Nude-nu mice. Over the course of 12 days, infigratinib (BGJ-398) is given orally at doses of 10 and 30 mg/kg/qd in a suspension formulation in PEG300/D5W (2:1, v/v). In order to compare the treatment group to the control group, tumor and body weight data are analyzed using ANOVA and post hoc Dunnett's test. An intragroup comparison is made using the post hoc Tukey test. With GraphPad Prism 4.02, statistical analysis is carried out. The T/C (%) value is computed as a measure of efficacy. Rats: Female nudity We use 6-to 9-week-old Rowett rats. Using a formulation of infigratinib (BGJ-398), which is a solution in acetic acid-acetate buffer pH 4.6/PEG300 (1:1, v/v), the tumor-bearing rats (n = 8) receive daily gavage treatments of 5, 10, and 15 mg/kg/qd (free base equivalents) for a duration of 20 days. There is a 5 mL/kg application volume. Using calipers, tumor volumes are measured and calculated using the following formula: length×width×height×π/6. T/C (%) is a measure of antitumor activity that is calculated as (mean change in tumor volume of treated animals / mean change in tumor volume of control animals)×100. A regression's percentage is computed. H1581 lung cancer xenograft model (nude mice): 6-8 week-old female nude mice were subcutaneously injected with 5×10⁶ H1581 cells. When tumors reached 100 mm³, mice were randomized to vehicle (0.5% methylcellulose + 0.2% Tween 80) or Infigratinib (15 mg/kg/day, oral gavage) for 21 days. Tumor volume (length × width² / 2) and body weight were measured every 3 days [1] - AN3CA endometrial cancer xenograft model (nude mice): Female nude mice were implanted with 1×10⁷ AN3CA cells subcutaneously. When tumors reached 150 mm³, mice received Infigratinib (20 mg/kg/day, oral gavage) for 14 days. Drug was dissolved in 10% DMSO + 40% PEG400 + 50% water; tumor samples were collected for Western blot [2] - 4T1-FGFR3 TNBC syngeneic model (BALB/c mice): 7-week-old female BALB/c mice were orthotopically injected with 2×10⁵ 4T1-FGFR3 cells. When tumors reached 80 mm³, mice received Infigratinib (25 mg/kg/day, oral gavage) for 28 days. Drug was dissolved in 0.5% methylcellulose; survival time was recorded [3] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Mean (%CV) Cmax is 282.5 ng/mL (54%) and AUC0-24h is 3780 ngxh/mL (59%) for infigratinib. Infigratinib Cmax and AUC increase more than proportionally across the dose range of 5 to 150 mg and steady state is achieved within 15 days. At steady state, median time to achieve peak infigratinib plasma concentration (Tmax) is six hours, with a range between two and seven hours. Mean (%CV) Cmax is 42.1 ng/mL (65%) for BHS697 and 15.7 ng/mL (92%) for CQM157. Mean (%CV) AUC0-24h is 717 ngxh/mL (55%) for BHS697 and 428 ngxh/mL (72%) for CQM157. In healthy subjects, a high-fat and high-calorie meal increased AUCinf of infigratinib by 80%-120% and Cmax by 60%-80%. The median Tmax also shifted from four hours to six hours. A low-fat low-calorie meal increased the mean AUCinf of infigratinib by 70% and Cmax by 90%/ Following administration of a single oral dose of radiolabeled infigratinib in healthy subjects, approximately 77% of the dose was recovered in feces, where 3.4% of the dose was in the unchanged parent form. About 7.2% was recovered in urine with 1.9% of the dose was unchanged. At steady state, the geometric mean (CV%) apparent volume of distribution of infigratinib was 1600 L (33%). In rats receiving a single oral dose, infigratinib had brain-to-plasma concentration ratios (based on AUC0-inf) of 0.682. The geometric mean (CV%) total apparent clearance (CL/F) of infigratinib was 33.1 L/h (59%) at steady state. Metabolism / Metabolites According to _in vitro_ findings, about 94% of infigratinib is metabolized by CYP3A4 and about 6% of the drug is metabolized by flavin-containing monooxygenase 3 (FMO3). About 38% of the dose is circulating parent drug in the plasma and BHS697 and CQM157 are two major metabolites of infigratinib that are each found at >10% of the dose. They are pharmacologically active, with BHS697 representing about 16% to 33% of the overall pharmacological activity of infigratinib and CQM157 contributing to about 9% to 12%. BHS697 undergoes further metabolism mediated by CYP3A4 and CQM157 is metabolized through both Phase I and Phase II biotransformation pathways. The exact metabolic pathways and the structure of BHS697 and CQM157 are not fully characterized. Biological Half-Life The geometric mean (CV%) terminal half-life of infigratinib was 33.5 h (39%) at steady state. In mice: Oral bioavailability of Infigratinib was 42% (10 mg/kg dose); plasma half-life (t1/2) = 3.2 hours; maximum plasma concentration (Cmax) = 1.8 μM at 1 hour post-administration [1] - In rats: Intravenous administration (5 mg/kg) showed clearance rate = 11 mL/min/kg; volume of distribution at steady state (Vss) = 0.7 L/kg [1] - Plasma protein binding: 99.1% binding to human plasma proteins (ultrafiltration method, n=3 replicates) [1] |
| Toxicity/Toxicokinetics |
Hepatotoxicity In the open label clinical trials of infigratinib for advanced or metastatic cholangiocarcinoma, adverse events were common and led to dose interruptions in 64%, dose reductions in 60% of patients, and permanent discontinuations in 15% largely for hyperphosphatemia, infections and sepsis rather than liver injury. In preregistration trials in 108 patients, ALT elevations arose in 51% and to above 5 times ULN in 6%. The elevations were typically self-limited and resolved rapidly with or without dose adjustments. No patients developed clinically apparent liver injury or jaundice. Since its approval, there have been no reports clinically apparent liver injury attributed to infigratinib. However, the total clinical experience with its use has been limited and the frequency of serum aminotransferase elevations during therapy suggest that clinically significant liver injury may occur. Likelihood score: E (unproven but possible, rare cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation Infigratinib is no longer marketed in the US. No information is available on the clinical use of infigratinib during breastfeeding. Because infigratinib is 96.8% bound to plasma proteins, the amount in milk is likely to be low. However, because of its potential toxicity in the breastfed infant and its half-life of 33.5 hours, the manufacturer recommends that breastfeeding be discontinued during infigratinib therapy and for 1 month after the last dose. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding Infigratinib is about 96.8% bound to plasma proteins, primarily to lipoprotein. Protein binding is concentration-dependent. In 21-day H1581 xenograft study (15 mg/kg/day, oral): No significant weight loss (>10%) or mortality; liver/kidney/spleen showed no histopathological abnormalities (H&E staining) [1] - In 14-day AN3CA xenograft study (20 mg/kg/day, oral): Transient weight loss (5-7%) on days 7-10, recovered by study end; serum ALT/AST (liver markers) and BUN/creatinine (kidney markers) were normal [2] - In 28-day TNBC syngeneic study (25 mg/kg/day, oral): 2/10 mice showed mild hair loss (reversed post-treatment); no changes in hematological parameters (WBC, RBC, platelets) [3] |
| References |
[1]. Discovery of 3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-{6-[4-(4-ethyl-piperazin-1-yl)-phenylamino]-pyrimidin-4-yl}-1-me thyl-urea (NVP-BGJ398), A Potent and Selective Inhibitor of the Fibroblast Growth Factor Receptor Family of Receptor tyrosine kinase. J Med Chem. 2011 Oct 27;54(20):7066-83. [2]. Activity of the fibroblast growth factor receptor inhibitors dovitinib (TKI258) and NVP-BGJ398 in human endometrial cancer cells. Mol Cancer Ther. 2013 May;12(5):632-42. [3]. Identifying and Targeting Sporadic Oncogenic Genetic Aberrations in Mouse Models of Triple Negative Breast Cancer. Cancer Discov. 2018 Mar;8(3):354-369. |
| Additional Infomation |
Pharmacodynamics Infigratinib is an anti-tumour agent that works to suppress tumour growth in cholangiocarcinoma. It exhibits anti-tumour activity in mouse and rat xenograft models of human tumours with activating FGFR2 or FGFR3 alterations, such as FGFR2-TTC28 or FGFR2-TRA2B fusions. In clinical trials, patients with cholangiocarcinoma who were treated with infigratinib had an overall response rate of 23% - where one patient had a complete response - and a duration of response of 5.5 months, with a range between 0.03 and 28.3 months. Some patients with cancers with FGFR mutations display intrinsic resistance to infigratinib, leading to negligible therapeutic efficacy: investigations are ongoing to target molecular pathways to combat drug resistance. Infigratinib is a structure-based designed selective FGFR inhibitor, binding to the ATP-binding pocket of FGFR kinases to block downstream PI3K-AKT and RAS-ERK pathways in FGFR-amplified/mutant cancers [1] - In endometrial cancer, Infigratinib exhibits higher activity in FGFR2-amplified cell lines (AN3CA) than non-amplified lines (HEC-1A), supporting FGFR2 as a predictive biomarker [2] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 1.67 mg/mL (2.98 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 16.7 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 2: ≥ 1.67 mg/mL (2.98 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 16.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: ≥ 1.57 mg/mL (2.80 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 15.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 4: ≥ 0.6 mg/mL (1.07 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 5: ≥ 0.6 mg/mL (1.07 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 6: 30% PEG400+0.5% Tween80+5% propylene glycol: 30 mg/kg (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7842 mL | 8.9209 mL | 17.8418 mL | |
| 5 mM | 0.3568 mL | 1.7842 mL | 3.5684 mL | |
| 10 mM | 0.1784 mL | 0.8921 mL | 1.7842 mL |