Neohesperidin dihydrochalcone is a synthetic glycoside chalcone, is added to various foods and beverages as a low caloric artificial sweetener.

Neohesperidin dihydrochalcone shows remarkable radical scavenging activity against stable radical and reactive oxygen species (ROS) in concentration dependent manner. Especially, neohesperidin dihydrochalcone is the most potent inhibitor of H₂O₂ and HOCl. Neohesperidin dihydrochalcone shows HOCl scavenging activity of 93.5% and H₂O₂ scavenging property of 73.5%. Neohesperidin dihydrochalcone shows extensive inhibitory effect especially on non-radical ROS H₂O₂ and HOCl with IC₅₀ values of 205.1, 25.5 μM^[1]. Neohesperidin dihydrochalcone is found to be an activator of porcine pancreatic alpha-amylase (PPA) with an IC₅₀ of 389 μM^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Neohesperidin dihydrochalcone administration results in significant reduction in activities of two useful markers of liver damage, AST and ALT. The relative levels of NF-κB, IL-6, IL-1β and TNF-α protein in the liver of PQ-treated mice are inhibited by neohesperidin dihydrochalcone^[3]. The embryotoxicity/teratogenicity of neohesperidin dihydrochalcone is examined in Wistar Crl:(WI)WU BR rats. No adverse effects are observed at neohesperidin dihydrochalcone levels of up to 5% of the diet, the highest dose level tested, at which the rats consumed about 3.3 g/kg body weight/day^[4].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

612.58

C₂₈H₃₆O₁₅

20702-77-6

新橙皮甙二氢查尔酮；新橙皮苷二氢查尔酮

Room temperature in continental US; may vary elsewhere.

DMSO : 100 mg/mL (163.24 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 (4.08 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (4.08 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 (4.08 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (4.08 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 (4.08 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (4.08 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]. Choi JM, et al. Antioxidant properties of neohesperidin dihydrochalcone: inhibition of hypochlorous acid-induced DNA strand breakage, protein degradation, and cell death. Biol Pharm Bull. 2007 Feb;30(2):324-30.

  [2]. Kashani-Amin E, et al. Neohesperidin dihydrochalcone: presentation of a small molecule activator of mammalian alpha-amylase as an allosteric effector. FEBS Lett. 2013 Mar 18;587(6):652-8.

  [3]. Shi Q, et al. Artificial sweetener neohesperidin dihydrochalcone showed antioxidative, anti-inflammatory and anti-apoptosis effects against paraquat-induced liver injury in mice. Int Immunopharmacol. 2015 Dec;29(2):722-9.

  [4]. Waalkens-Berendsen DH, et al. Embryotoxicity and teratogenicity study with neohesperidin dihydrochalcone in rats. Regul Toxicol Pharmacol. 2004 Aug;40(1):74-9.

WST-8 dye is used in the cell viability assay. HIT-T15 and HUVEC cells are grown and maintained in Dulbecco’s modified Eagle’s medium, supplemented with 10% fetal bovine calf serum. 1000 cells in each well are incubated with various concentrations of neohesperidin dihydrochalcone (50, 100, 500 μM, 1 mM) and other compounds. After treating HIT-T15 and HUVEC cells with 500 μM HOCl, WST-8 dye is added to each well, and the absorbance is detected at 420 nm with microplate reader^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Rats: The embryotoxicity/teratogenicity of neohesperidin dihydrochalcone is examined in Wistar Crl:(WI)WU BR rats. The study is comprised of four groups of 28 mated female rats each, i.e., a control group (0% neohesperidin dihydrochalcone) and three treatment groups (1.25, 2.5, and 5% neohesperidin dihydrochalcone). The general condition and behavior of the animals are observed twice daily. Body weight is determined on days 0, 7, 14, and 21 of gestation. Food consumption is determined during three consecutive periods (days 0-7, 7-14, and 14-21 of gestation)^[4].

^[3]Mice: Neohesperidin dihydrochalcone is dissolved in a 0.5% CMC vehicle. Mice are randomized into four groups. The control group receives equal volume of vehicles throughout. The PQ group receives saline once daily for 6 consecutive days. One hour after final saline treatment, mice are injected with PQ (75 mg/kg body weight). The neohesperidin dihydrochalcone group receives a daily dose of 200 mg/kg body weight by oral gavage for 6 consecutive days. One hour after final neohesperidin dihydrochalcone treatment, mice are injected with PQ (75 mg/kg body weight)^[3].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Choi JM, et al. Antioxidant properties of neohesperidin dihydrochalcone: inhibition of hypochlorous acid-induced DNA strand breakage, protein degradation, and cell death. Biol Pharm Bull. 2007 Feb;30(2):324-30.

  [2]. Kashani-Amin E, et al. Neohesperidin dihydrochalcone: presentation of a small molecule activator of mammalian alpha-amylase as an allosteric effector. FEBS Lett. 2013 Mar 18;587(6):652-8.

  [3]. Shi Q, et al. Artificial sweetener neohesperidin dihydrochalcone showed antioxidative, anti-inflammatory and anti-apoptosis effects against paraquat-induced liver injury in mice. Int Immunopharmacol. 2015 Dec;29(2):722-9.

  [4]. Waalkens-Berendsen DH, et al. Embryotoxicity and teratogenicity study with neohesperidin dihydrochalcone in rats. Regul Toxicol Pharmacol. 2004 Aug;40(1):74-9.