Trisodium citrate exhibits dose-dependent antiproliferative action (0–12.5 mM; 24 hours) [3]. In G2/M and S phases, trisodium citrate (12.5 mM; 72 h) dose-dependently promotes apoptosis and cell cycle arrest [3]. The expression of FAS, BAX, BID, AIF, EndoG, cytochrome c, PARP, GADD153, GRP78, and caspase-3, -8, and -9 was upregulated and the expression of BCL-2 and BCL-Xl was downregulated after 48 hours at 12.5 mM of trisodium citrate[3].

In mouse livers, intraperitoneal injections of trisodium citrate (120, 240, and 480 mg/kg) can dramatically lower GSH-Px activity and raise MDA (malondialdehyde) levels [1]. Mouse hepatocytes exposed to intraperitoneal trisodium citrate (120, 240, and 480 mg/kg) exhibit dose-dependent increases in caspase-3 activity, which results in apoptosis [1]. Mice exposed to intraperitoneal injections of trisodium citrate (120, 240, and 480 mg/kg; once weekly for three weeks) develop nephrotoxicity [2].

Cell Viability Assay[3]
Cell Types: HaCaT Cell
Tested Concentrations: 0, 2.5, 5, 7.5, 10, 12.5 mM
Incubation Duration: 24 hrs (hours)
Experimental Results: Inhibition of cell viability in a dose-dependent manner.

---

Cell cycle analysis[3]
Cell Types: HaCaT Cell
Tested Concentrations: 12.5 mM
Incubation Duration: 0, 12, 24, 48, 72 hrs (hours)
Experimental Results: Induced apoptosis and cell cycle arrest in G2/M phase and S in a dose-dependent manner Expect.

---

Western Blot Analysis[3]
Cell Types: HaCaT Cell
Tested Concentrations: 12.5 mM
Incubation Duration: 12, 24, 48 hrs (hours)
Experimental Results: Increased expression of FAS, BAX, BID, AIF, EndoG, cytochrome c, PARP, GADD153, GRP78 and caspase -3, -8, -9, and BCL-2 and BCL-X1 were diminished.

Animal/Disease Models: 20 g male Kunming mice [2]
Doses: 120, 240, 480 mg/kg
Route of Administration: intraperitoneal (ip) injection; once a week for 3 weeks.
Experimental Results: The activities of T-SOD and GSH-Px in the treatment group diminished with the increase of citric acid dose, the activity of NOS demonstrated an increasing trend, and the contents of H2O2 and MDA gradually diminished.

Absorption, Distribution and Excretion
Time to Peak (Tmax): 98-130 minutes. Primarily eliminated through hepatic metabolism; minimal renal clearance. 19-39 liters. Total clearance: 313-1107 ml/min. In vivo, sodium citrate is oxidized to bicarbonate and excreted in urine… Metabolism/Metabolites Citrate is metabolized to bicarbonate in the liver and acts as an intermediate in the citric acid cycle. In vivo, sodium citrate is oxidized to bicarbonate… Biological Half-Life: 18-54 minutes.

[1]. Study on injury effect of food additive citric acid on liver tissue in mice. Cytotechnology. 2014 Mar;66(2):275-82.

[2]. Chen X, Lv Q, Liu Y, Deng W. Effects of the food additive, citric acid, on kidney cells of mice. Biotech Histochem. 2015 Jan;90(1):38-44.

[3]. Citric acid induces cell-cycle arrest and apoptosis of human immortalized keratinocyte cell line (HaCaT) via caspase- and mitochondrial-dependent signaling pathways. Anticancer Res. 2013 Oct;33(10):4411-20.

Sodium citrate is the trisodium salt of citric acid. It is used as a flavoring agent and anticoagulant. It contains citrate (3-). Sodium citrate is the sodium salt of citric acid. It is a white crystalline powder or white granular crystal, slightly deliquescent in moist air, readily soluble in water, and almost insoluble in alcohol. Like citric acid, it has a sour taste. From a medical perspective, it is used as an alkalizing agent. It works by neutralizing excess acid in the blood and urine. It has been used to treat metabolic acidosis. Sodium citrate is the sodium salt of citrate, which has an alkalizing effect. After absorption, sodium citrate dissociates into sodium ions and citrate anions; the organic citrate ions are metabolized into bicarbonate ions, leading to an increase in plasma bicarbonate concentration, buffering excess hydrogen ions, raising blood pH, and potentially reversing acidosis. Furthermore, taking sodium citrate can increase free sodium load, thereby increasing intravascular blood volume, promoting the excretion of bicarbonate compounds, and exerting an anti-calculi effect. Sodium citrate is used as a buffer and food preservative. Medically, it is used as an anticoagulant for storing blood and as a urine alkalizer to prevent kidney stones. See also: Anticoagulant sodium citrate solution (with subclasses); Sodium chloride; Sodium citrate, unspecified form (ingredient)... See more...

---

Drug Indications
Used as an anticoagulant during plasma exchange and as a neutralizer for treating stomach upset and acidic urine.
FDA Label

---

Mechanism of Action
Citrate chelates free calcium ions, preventing them from forming complexes with tissue factor and coagulation factor VIIa, thereby inhibiting the activation of coagulation factor X. This inhibits the extrinsic initiation of the coagulation cascade. Citrate may also exert its anticoagulant effect through a currently unknown mechanism, as restoring calcium concentration does not completely reverse the effects of citrate. Citrate is a weak base and therefore reacts with hydrochloric acid in the stomach, raising the pH. It is further metabolized into bicarbonate, which then acts as a systemic alkalizing agent, raising the pH of blood and urine. It also has a diuretic effect and increases the excretion of calcium in urine.

C6H5NA3O7

258.0690

257.972

68-04-2

Citric acid;77-92-9

6224

White to off-white solid powder

1.008 g/mL at 20 °C

300°C

1

7

2

16

211

0

HRXKRNGNAMMEHJ-UHFFFAOYSA-K

InChI=1S/C6H8O7.3Na/c7-3(8)1-6(13,5(11)12)2-4(9)10;;;/h13H,1-2H2,(H,7,8)(H,9,10)(H,11,12);;;/q;3*+1/p-3

trisodium;2-hydroxypropane-1,2,3-tricarboxylate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

H2O : ~50 mg/mL (~193.75 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

---

Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

View More

Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

---

Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

View More

Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

---

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

---

 (Please use freshly prepared in vivo formulations for optimal results.)