Lucidin (1,3-dihydroxy-2-hydroxymethyl-9,10-anthraquinone), was mutagenic in five Salmonella typhimurium strains without metabolic activation, but the mutagenicity was increased after addition of rat liver S9 mix. In V79 cells, lucidin was mutagenic at the hypoxanthine-guanine phosphoribosyl transferase gene locus and active at inducing DNA single-strand breaks and DNA-protein cross-links as assayed by the alkaline elution method. Lucidin also induced DNA repair synthesis in primary rat hepatocytes and transformed C3H/M2-mouse fibroblasts in culture. We also investigated lucidinethylether, which is formed from lucidin by extraction of madder roots with boiling ethanol. This compound was also mutagenic in Salmonella, but only after addition of rat liver S9 mix. Lucidinethylether was weakly mutagenic to V79 cells which were cocultivated with rat hepatocytes. The compound did not induce DNA repair synthesis in hepatocytes from untreated rats, but positive results were obtained when hepatocytes from rats pretreated with phenobarbital were used. We conclude that lucidin and its derivatives are genotoxic.[1]

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lucidin, a hydroxyanthraquinone derivative present in this plant, is mutagenic in bacteria and mammalian cells. We also demonstrated the formation of DNA adducts in tissue culture and mice after treatment with this compound. To elucidate the possible carcinogenicity of madder root, three groups of male and female ACI rats received either a normal diet or a diet supplemented with 1 or 10% drug for a total period of 780 days. Weight gain and morbidity were not different among the three groups. Non-neoplastic lesions related to the treatment were evident in the liver and kidneys of both sexes. Moreover, dose-dependent increases in benign and malignant tumour formation were observed in the liver and kidneys of treated animals. 32P-post-labelling analysis showed an increase in the overall level of DNA adducts observed in the liver, kidney and colon of rats treated with 10% madder root in the diet for 2 weeks. HPLC analysis of 32P-labelled DNA adducts revealed a peak co-migrating with an adduct obtained after in vitro treatment of deoxyguanosine-3'-phosphate with lucidin. These observations suggest that the use of madder root for medicinal purposes is associated with a carcinogenic risk.[2]

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Lucidin was mutagenic in five Salmonella typhimurium strains (TA 98, TA 100, TA 102, TA 104, TA 1537, TA 1538) without metabolic activation, with mutagenicity increased upon addition of rat liver S9 mix. [1]
Lucidin induced dose-related increases in 8-azaguanine-resistant colonies in V79 Chinese hamster fibroblasts at concentrations of 5–30 µg/ml, indicating mutagenicity at the HGPRT locus. [1]
Lucidin induced DNA repair synthesis (unscheduled DNA synthesis, UDS) in primary rat hepatocytes, with significant increases in net grains/nucleus at concentrations ≥6.3 µg/ml. [1]
Lucidin induced transformed foci in C3H/M2 mouse fibroblasts in culture, indicating malignant transformation potential. [1]
Lucidin induced DNA single-strand breaks and DNA-protein cross-links in V79 cells as detected by the alkaline elution method. [1]

Long-term dietary administration of madder root (containing Lucidin) to ACI rats induced dose-dependent increases in benign and malignant tumors in the liver and kidneys. [2]
Lucidin derived from madder root formed DNA adducts in the liver, kidney, and colon of rats after 2 weeks of dietary administration (10% madder root). [2]

V79 HGPRT Mutagenicity Assay: V79 Chinese hamster fibroblasts were treated with Lucidin at concentrations ranging from 5 to 30 µg/ml. After treatment, cells were plated in medium containing 8-azaguanine to select for HGPRT-deficient mutants. The number of resistant colonies was counted and normalized to plating efficiency. [1]
Primary Rat Hepatocyte DNA Repair (UDS) Assay: Hepatocytes were isolated from male Wistar rats and cultured. Cells were exposed to Lucidin at various concentrations, then incubated with ³H-thymidine. DNA repair synthesis was assessed autoradiographically by counting net grains per nucleus. [1]
C3H/M2 Mouse Fibroblast Transformation Assay: C3H/M2 cells were treated with Lucidin at concentrations from 1 to 10 µg/ml. After treatment, cells were cultured and monitored for the formation of morphologically transformed foci. [1]
Alkaline Elution Assay for DNA Damage: V79 cells were treated with Lucidin for 60 minutes. DNA single-strand breaks and DNA-protein cross-links were analyzed by alkaline elution on polycarbonate filters, using tritium-labeled reference cells irradiated with 3000 RAD as a control. [1]

Long-term Carcinogenicity Study: ACI rats (both sexes, 150-200 g) were divided into three groups: control (standard diet), low dose (diet supplemented with 1% madder root), and high dose (diet supplemented with 10% madder root). The madder root was pulverized and mixed into the diet pellets. Animals had free access to food and water. The study duration was 666-781 days. Body weight was monitored weekly. Moribund animals or those with marked weight loss were euthanized. At study termination, all remaining animals were euthanized under deep ether anesthesia for necropsy and tissue collection. [2]
DNA Adduct Formation Study: A separate group of rats was fed a diet containing 10% madder root for 2 weeks. After this period, DNA was isolated from liver, kidney, and colon for ³²P-post-labeling analysis of DNA adducts. [2]

In animal studies, the average daily intake of Lucidin was calculated based on its content in the diet: the high-dose group (10% madder feed) had a daily intake of approximately 33.6 mg of Lucidin per kilogram of body weight; the low-dose group (1% madder feed) had a daily intake of approximately 3.36 mg of Lucidin per kilogram of body weight. [2]

Lucidin showed toxicity against Salmonella typhimurium strain TA 1535 at high doses, as indicated by the "tox" marker in the reverse mutation count table. [1] DNA damage (strand breaks and cross-links) in the alkaline elution assay indicated its potential genotoxicity. [1]

[1]. The genotoxicity of lucidin, a natural component of Rubia tinctorum L., and lucidinethylether, a component of ethanolic Rubia extracts. Cell Biol Toxicol. 1988 Jun;4(2):225-39.

[2]. Carcinogenicity and DNA adduct formation observed in ACI rats after long-term treatment with madder root, Rubia tinctorum L. Carcinogenesis. 1998 Dec;19(12):2163-8.

Lucidin is a dihydroxyanthraquinone. It has been reported that Lucidin exists in Rubia argyi, Rubia wallichiana and other organisms with relevant data. Mechanism of action Lucidin is a mutagenic anthraquinone found in Rubiaceae plants. Under physiological conditions, it reacts with nucleic acid bases to form adducts. ... The isolated purine base adduct was identified as a reactant formed by the condensation of the benzylic position of compound 1 with the nitrogen atom of the purine base. Lucidin is a natural hydroxyanthraquinone component in the root of Rubia tinctorum L. [1] It is mutagenic in bacterial and mammalian cell systems, can induce DNA damage and repair, and can cause malignant transformation of mouse fibroblasts in vitro. [1] Lucidin can be converted into Lucidin ether (LUE) by boiling ethanol extraction of Rubia argyi root. The latter also exhibits mutagenicity after metabolic activation. [1]
This study suggests that treatment with madder extract containing Lucidin may pose a potential carcinogenic risk to humans. [1]

C15H10O5

270.2369

270.052

478-08-0

478-08-0 (Lucidin); 17526-17-9 (Lucidin ethyl ether)

10163

Light yellow to yellow solid powder

1.6±0.1 g/cm3

585.0±29.0 °C at 760 mmHg

330ºC

321.6±20.8 °C

0.0±1.7 mmHg at 25°C

1.746

2.79

3

5

1

20

421

0

AMIDUPFSOUCLQB-UHFFFAOYSA-N

InChI=1S/C15H10O5/c16-6-10-11(17)5-9-12(15(10)20)14(19)8-4-2-1-3-7(8)13(9)18/h1-5,16-17,20H,6H2

1,3-Dihydroxy-2-(hydroxymethyl)anthracene-9,10-dione

NSC 30546; NSC30546; NSC-30546; Lucidin; Henine;

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 : ~6.67 mg/mL (~24.68 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.

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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).

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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)

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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).

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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

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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.

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