Citric acid is a weak organic tricarboxylic acid found in citrus fruits. Citric acid is a natural preservative and food tartness enhancer.

Citric acid induces apoptosis through the mitochondrial pathway in the human keratinocyte cell line HaCaT. It inhibits proliferation of HaCaT cells in a dose-dependent manner, but also induces apoptosis and cell cycle-arrest at the G2/M phase (before 24 h) and S phase (after 24 h)^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Citric acid is found in all animal tissues as an intermediary substance in oxidative metabolism. The administration of citric acid (1–2 g/kg) attenuates LPS-induced elevations in brain MDA, nitrite, TNF-α, GPx, and PON1 activity. In the liver, nitrite is decreased by 1 g/kg citric acid. Citric acid (1-2 g/kg) decreases brain lipid peroxidation and inflammation, liver damage, and DNA fragmentation^[2]. Citric acid supplementation increases intestinal calcium and phosphorus absorption and the retention/intake ratio only in rats fed the 1% Ca diet. Citric acidsupplementation together with a calcium-rich diet allows to obtain an increased retention of calcium and phosphorus in bone. The prolonged administration of calcium citrate supplements may therefore help to increase bone mineral concentration^[3]. Oral administration of citric acid ameliorates ketosis and protects against the development of diabetic complications in an animal model of type 1 diabetes^[4].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

192.12

C₆H₈O₇

77-92-9

柠檬酸

Room temperature in continental US; may vary elsewhere.

H₂O : 100 mg/mL (520.51 mM; Need ultrasonic)

DMSO : 100 mg/mL (520.51 mM; Need ultrasonic)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.5 mg/mL (13.01 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (13.01 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.5 mg/mL (13.01 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (13.01 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (13.01 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (13.01 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Ying TH, et al. 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.

  [2]. Abdel-Salam OM, et al. Citric acid effects on brain and liver oxidative stress in lipopolysaccharide-treated mice. J Med Food. 2014 May;17(5):588-98.

  [3]. Lacour B, et al. Stimulation by citric acid of calcium and phosphorus bioavailability in rats fed a calcium-rich diet. Miner Electrolyte Metab. 1997;23(2):79-87.

  [4]. Nagai R, et al. Citric acid inhibits development of cataracts, proteinuria and ketosis in streptozotocin (type 1) diabetic rats. Biochem Biophys Res Commun. 2010 Feb 26;393(1):118-22.

HaCaT cells are treated with different concentrations of citric acid (2.5, 5, 7.5, 10, 12.5 mM) for 24 h; 0.5% of DMSO (vehicle) is used as a control. Cells are then centrifuged at 1000 ×g for 5 min, and cell pellets are dissolved with 0.5 mL of Phosphate buffered saline (PBS) containing 5 μg/mL PI and viable cells are determined by using a flow cytometer for determination of viable cells^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Rats: Rats are induced of diabetes and divided randomly into an untreated diabetic group and two treated diabetic groups, receiving citric acid (2 g/L) in the drinking water, or insulin therapy. Insulin-treated rats receive 3 IU of neutral protamine Hagedorn insulin three times per week. Fasting (12 hr) blood samples are obtained from the tail vein two days after insulin injection for measurement of blood glucose, HbA1c and ketone bodies^[4].

^[2]Mice: Citric acid is prepared in sterile physiological saline. Mice are randomly divided into five equal groups (six mice each). Mice are treated with either 0.2mL of: sterile physiological saline (group 1) or citric acid at doses of 1, 2, and 4 g/kg, orally (groups 2-4). Treatments are given just prior to endotoxin administration (LPS: 200 lg/kg, injected intraperitoneally, 0.1 mL). The fifth group received just the vehicle, no LPS (negative control). Mice are euthanized after 4 h of LPS or vehicle injection by decapitation under ether anesthesia, where the brain and liver of each mouse are then removed for analysis^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Ying TH, et al. 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.

  [2]. Abdel-Salam OM, et al. Citric acid effects on brain and liver oxidative stress in lipopolysaccharide-treated mice. J Med Food. 2014 May;17(5):588-98.

  [3]. Lacour B, et al. Stimulation by citric acid of calcium and phosphorus bioavailability in rats fed a calcium-rich diet. Miner Electrolyte Metab. 1997;23(2):79-87.

  [4]. Nagai R, et al. Citric acid inhibits development of cataracts, proteinuria and ketosis in streptozotocin (type 1) diabetic rats. Biochem Biophys Res Commun. 2010 Feb 26;393(1):118-22.