Science topic

Anthocyanins - Science topic

A group of flavonoids derived from flavonols, which lack the ketone oxygen at the 4-position. They are glycosylated versions of cyanidin, pelargonidin or delphinidin. The conjugated bonds result in blue, red, and purple colors in flowers of plants.
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Hello, please I need help with a statistical analysis. I don't know what statistical test to use. The study I carried out is as follows: Quantify polyphenols, flavonoids and anthocyanins in addition to antioxidant activity for 2 different concentrations of a fruit. I'm not sure what statistical analysis to perform
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You should use a randomized complete block design, 2×4× reps. You should as much replications as you can to increase prescion of your results
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I am sampling tree leaves for pigment analysis with a UV-Vis Spectrometer. The samples will be snap-frozen in the field and stored in the dark. I am trying to determine the best method for anthocyanin extraction and analysis. It would also be great to know if it could be integrated into a chlorophyll and carotenoid extraction too? Thanks in advance!
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Dear Procopio Peinado-Torrubia and J.D. Franco-Navarro , thank you for your advice! If it is possible to be linked to the protocols you discussed above I would really appreciate it.
Many Thanks,
Jack
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I read an article about the optimization of anthocyanin, flavonol and phenolic acid extractions.
I wondered if these extracts would damage the colon when run with LC or not.
Adjé, F., Lozano, Y. F., Lozano, P., Adima, A., Chemat, F., & Gaydou, E. M. (2010). Optimization of anthocyanin, flavonol and phenolic acid extractions from Delonix regia tree flowers using ultrasound-assisted water extraction. Industrial Crops and Products, 32(3), 439-444.
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Dear Dr.Burnaz
Why use sulfuric acid to extract?
In Green Chemistry exist many solvents such as carboxylic acids; glycerol; etc. Nesibe Arslan Burnaz
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I stored at sample 25 °C for 2 weeks. Absorbance was read at 520 nm after 0 and 48 h, 7 and 14 days.
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You can calculate it as a loss of colour strength ( absorbance of 1% sol.) as a function of time.
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For my thesis, I need additional information, numbers, raw materials about the content of some anthocyanins, total phenolic composition, or antioxidant activity in grape seed extract in natural and after fermentation conditions. If you have any of them, please, send me.
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Hope this review can help.
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I have been trying to extract anthocyanin using different concentrations of alcohol. For that TPC, TAC and Antioxidant activity (DPPH method) are the following analysis that I'm performing.
So, the different method of extractions (different concentrations) shows different results.
For e.g.,
Suppose, Method 1 has indicated more antho content but the antioxidant activity is less.
Method 2 shows less antho or may be less TPC but more antioxidant activity.
So my question is
What should be the criteria for the selection of a method as i want both more TAC value and antioxidant content?
I'm highly confused , please help.
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so i got the peak for cyanidin glucoside correspond to the reference standard but i have other peaks in my samples how can i identify them?
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Does anyone know a reliable method to measure anthocyanin content with spectrophotometer?
I am working on Tradescantia Pallida leaves.
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Hi,
I am looking for good articles on Dye-Sensitized Solar Cells (DSSC) by which one with a minimum knowledge about this subject can quickly understand how DSSC works.
Specially articles fully describing the function of dyes like Anthocyanin in DSSC are welcomed.
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Is the reaction of Anthocyanin to pH reversible? I mean if we increase the pH and get the color change, if we then decrease the pH, the color will be changed back to its original color?
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I also recemmend you to read these two interesting articles (in attached).
Best wishes,
Sabri
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Hi there,
i was wondering if someone could help me with my question.
i am working on drying of fruits and vegeatables.
4.5 g of fresh sample after drying process will reduces to 0.570 g.
For anthocyanin, phenolic content and antioxidant capacity i add 1g of dried sample to 10 ml of solvent.
how much of fresh sample should be added ?
If any citeable source please mention.
Your help is much appreciated.
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Absorbance (A) = (Aλ vis-max x A700) pH 1.0 0 (Aλ vis-max x A700) pH 4.5
TAC (mg/L) = (A × MW × DF × 1000)/(ε × l)
MW: the molecular weight, calculated as cyanidin-3-glucoside (449.2); DF: the dilution factor;
l: the cuvette radius, 1 cm; ε: the molar absorptivity, calculated as cyanidin-3-glucoside (26,900).
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For justification of results it is necessary to run reference standard.
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Hello. Please i want to know how to prepare standard (cyanidin-3- glucoside) for determining total anthocyanin using pH differential method ?
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Dear sir
The total anthocyanin content (TAC) was determined by the pH-differential method.[12] 1 ml of TAE solution (1 mg/ml) was mixed separately with 9 ml buffer at pH 1.0 (0.1 M HCl/4.9 mM KCl) and another at pH 4.5 (24.8 mM sodium acetate), and then the mixtures were balanced for 1 h in the dark. Absorbance was measured in a WFZ UV-2000 UV and visible spectrophotometer at 510 nm and 700 nm in buffers of pH 1.0 and pH 4.5, respectively. TAC was expressed as milligrams cyanidin-3-glucoside equivalents per gram of dry weight purification (mg C-3-G/g DW) and calculated via the following formula:
you can read
Total anthocyanins and cyanidin-3-O-glucoside contents and antioxidant activities of purified extracts from eight different pigmented plants
DOI: 10.4103/pm.pm_162_18
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I don't know if we have a freeze-dryer or spray-dryer here in our area that's why I'm opting to use Ionic gelation, is it an effective method? or can you recommend a more effective method that don't require expensive instruments?
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Many plants produced various classes of compounds such as polyphenols (phenolic acids, flavonoids, anthocyanins, lignans and stilbenes), carotenoids (xanthophylls and carotenes) and vitamins (vitamin E and C) which have potent antioxidant activity.
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Polyphenols and carotenoids are molecules that have potent antioxidant activity because they sometimes act synergistically in the oxidation system, where polyphenols act as scavengers, while carotenoids act as chelators of free radicals.
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Blood oranges have a reddish orange outer peel and red color flesh, the red color comes from anthocyanins, which are not only pigments but powerful antioxidants with many health benefits, different studies have shown an association between anthocyanins’ high antioxidant activity and a decreased risk of chronic diseases such as cancer, heart disease and type 2 diabetes.
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Blood oranges are full of anthocyanins, a type of antioxidant. Numerous studies have shown an association between anthocyanins’ high antioxidant activity and a decreased risk of chronic diseases such as cancer, heart disease, and type 2 diabetes.
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As weighing anthocyanin standard (keracyanin chloride; ACN STD) is very difficult and risky, the plan is to dissolve ACN STD with analytical solvent which is not soluble with water or alcohol (Information source: https://www.scbt.com/p/keracyanin-chloride-18719-76-1#:~:text=In%20other%20studies%2C%20Keracyanin%20chloride,through%20inhibition%20of%20α-glucosidase.&text=Solubility%20%3A,Soluble%20in%20water%2C%20and%20alcohol.); thus, what solvent is suitable for dissolving ACN STD without contaminating it?
After that, the desiccant is used to dry analytical solvent; thus, what desiccant is suitable without contaminating ACN STD?
Link of research papers or video from official source will be helpful, but it's optional.
This question is related to 'How to transfer ACN STD correctly?' previously, and a video demo can be watched for better understanding.
Thank you.
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I want to know the genetics of Black rice, if anyone had published one,or have any articles please share to me.
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The anthocyanin standard is a keracyanin chloride and it is used for making standard curve of anthocyanin concentration. Its characteristic is hygroscopic (very sticky); thus, it is not possible to use regular spatula due to very small plastic microtube. By using needle, it is still difficult to transfer the powder. Besides, the solvent to be used has very low pH (the anthocyanin analysis method used is pH Differential Method), so it is not recommended to dilute in microtube by using micropipette (the tips are made of plastic). For better understanding, attached are video of demonstration and protocols for making anthocyanin standard. Hopefully, the problem will be solved. Thank you
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May 10, 2021
Dear Augusto,
Your video shows that the vial contains 5 mG of keracyanin-Cl. As you learned, it is very hygroscopic. This makes it impossible to weigh it out accurately, as it sticks to the spatula, Al-foil, etc.
If I were you, I wouldn't try weighing it out. Even if you could do this, since the material is hygroscopic, it would absorb H20 from the air during the weighing step. As such, the weight would be inaccurate due to the absorbed moisture. Instead, add a known volume of your solvent to the vial. Cap it tightly and mix it gently until everything dissolves. You can even add a small magnetic stirring bar ("flea")to the vial and put in on a stir plate until solution is complete. From the AOAC method, the material is exposed to pH 1.0. This will not harm the plastic vial or plastic pipet tips.
Once the material completely dissolves, you can transfer it to a volumetric flask and calculate the concentration from the material weight and final solvent volume. You should get the Certificate of Analysis (COA) for this material from Sigma's website and check the product purity. If the material is <100% pure, you will need to take this into account to calculate the concentration accurately. To get the COA, you will need to know the Lot # for this product. This is shown on the vial's label.
If you have any other questions about this material, you can contact Sigma directly by e-mail. I've used Sigma's products in the past and have contacted them many times with questions about their chemicals. They are always willing to help and very good at providing information.
I hope this information helps you.
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Hello every one please can any one help me for procedure/formulae on how to calculate anthocyanin biosynthesis related such as 4CL, C4H and CHS ?
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What HPLC Marker(s) or standard is used to detect and quantify anthocyanins such as Delphinidin, Cyanidin in a given sample of plant extract?
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Dear Yahyea Baktiar Laskar thank you for this interesting question. Please see e.g. the following potentially useful article which might be helpful in answering your question:
1. Recent Advances in Anthocyanin Analysis and Characterization
and
2. Identification of Anthocyanin Composition and Functional Analysis of an Anthocyanin Activator in Solanum nigrum Fruits
The first article is freely available as public full text on RG, while the second paper is Open Access and can be downloaded for free from the general internet. Please find attached a pdf file of this article.
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Hello everyone. I do my anthocyanin test for biscuit incorporated with muntingia calabura fruit powder.
I use ultrasonic bath for extraction biscuit powder in metanol HCl 0,1 N (1:10, w/v), filtered, and centrifuged, then collected the supernatan for analysis.
I use potasium chloride buffer (pH= 1) and sodium citric acid buffer, instead of acetate.
I got turbidity solution. Does anyone know where the turbidity come from?
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Anthocyanins are more stable at low pH (acidic conditions) which gives a red pigment. Meanwhile, the higher the pH value of anthocyanin will provide color fading of the color blue. So as a food colorant, anthocyanin with a low pH or height pH has a significant effect on the food colorant.
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Hello everybody!
I have performed an acidic MeOH:HCl 1% (v/v) plant extraction from Arabidopsis leaves to calculate Anthocyanins concentrations based on:
Rabino, I., and Mancinelli, A. L. 1986. Light, temperature, and anthocyanin production. Plant Physiol. 81:922-924.
I want to calculate also Chl a, Chl b and Carotenoids concentrations from this acidic methanolic extraction. So far, I have only found 100% MeOH equations to calculate concentrations of these pigments, separately, but not a single reference to calculate them from a MeOH:HCl 1% (v/v) plant extract.
I have read that "A657 values provide an estimate of Chl content" (Photoregulation of Anthocyanin Synthesis' VIII. EFFECT OF LIGHT PRETREATMENTS, Mancinelli, 1983), but I would like to have a more precise value for Chl a, Chl b, and carotenoids also, if possible.
I have recorded measurements of wavelengths between 300-700 nm.
I would be grateful if anyone helps me with that.
Thank you all!
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Hi, you can find more information in this chapter (Chlorophylls and Carotenoids: Measurement and Characterization by UV‐VIS Spectroscopy by Hartmut K. Lichtenthaler, Claus Buschmann). There are formulas for calculating the concentration of chlorophyll (a; b) and total carotenoids (x + c), depending on the solvent used in the extraction.
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I use the water method to extract the anthocyanidin from Peristrophe bivalvis (L.) Merr leave, then dilute 20 times with 0.025M potassium chloride, pH1.0 and 0.4M sodium acetate buffer, pH4.5 respectively. To measure the absorbance at 510nm and 700nm.
Set into the formula:
Anthocyanin pigment concentration(mg/liter)=(A*MW*DF*1000)/(ɛ*1) \
But now I have a problem is that the result of the A=(A510-A700)pH1.0-( A510-A700)pH4.5 is a negative value that I can’t have result from this formula.
So my question is Why A would be a negative value?
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I use the 1.5N HCl/EtoH method to extract the anthocyanidin from potato roots, then dilute 5 times with 0.025M potassium chloride, pH1.0 and 0.4M sodium acetate buffer,pH4.5 respectively. To measure the absorbance at 510nm and 700nm.
Set into the formula: Anthocyanin pigment concentration(mg/liter)=(A*MW*DF*1000)/(ɛ*1)
But now I have a problem is that the result of the A=(A510-A700)pH1.0-( A510-A700)pH4.5 is a negative value that I can’t have result from this formula.
So my question is Why A would be a negative value?
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Dear Chiao-Yi Chang,
I have the same problem (negative A) when I used pH-different method for Peristrophe bivalvis (L.) Merr leave. You already solved the problem or not? How do you do? Could you give me the advice?
Thank you in advance!
Ngo Van Tai, from Vietnam.
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I would like to know more about the light stability test for anthocyanin using UV light and agar solution. If we use agar solution, how can we read absorbance value? Or else can we show the light stability of anthocyanin in color measured with tintometer. Thank you all in advance.
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The Tintometer colour measurement instrument is made by the Lovibond Company in Aylesbury Wiltshire UK and uses visual standard comparison to quantify colour. Coordinate colour and colour difference definitions in Cielab Cluv and CIE XYZ colour space, covering both light stability (i.e. colour fastness) and AOC Light Absorbance scale are supported. 
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i am working on anti-viral drug designing so i need a detail list of flavonoids only not its sub group i-e
  • Anthocyanidins
  • 3.2Anthoxanthins
  • 3.3Flavanones
  • 3.4Flavanonols
3.5Flavan
so if someone can provide me this list with atleast 100 flavonoids and their derivatives .
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Flavonoids related to immuno system its act as antioxidants
Antimicrobial or antifubgal its ok
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I'm trying to quantify the anthocyanins in tomato leaves using LC-MS/MS. The two most common anthocyanins in tomato vegetative tissue are petunidin-3-(p-coumaryl rutinoside)-5-glucoside and malvidin-3-(p-coumaryl rutinoside)-5-glucoside.
Is it possible to quantify these two anthocyanins with the commercially available standard malvidin-3-glucoside using molecular-weight correction factors as in the following article?
( Chandra, A., Rana, J., & Li, Y. (2001). Separation, identification, quantification, and method validation of anthocyanins in botanical supplement raw materials by HPLC and HPLC− MS. Journal of agricultural and food chemistry, 49(8), 3515-3521.)
Or is it better to quantify petunidin-3-(p-coumaryl rutinoside)-5-glucoside using petunidin-3-glucoside as a standard?
Is it possible that malvidin-3-(p-coumaryl rutinoside)-5-glucoside and the standard malvidin-3-glucoside don't have the same retention time and is this a problem for the quantification?
Thank you for the help!
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Hi, I think it is possible, but at first you should determine relative conversion factor.
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Almost all publications use same formula to determine the total anthocyanin content. As you know, generally molar extinction coefficient of cyanidin-3-glucoside is 29600 in this formula. Differently, can i use calibration curve of cyanidin-3-glucoside or different anthocyanin standard to determine this.
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I am looking for available databases for a standardized list of SOPs. More specifically, I am looking for a standardized SOP for anthocyanin extraction from deepwater rice. This SOP must be for industrial use.
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In the protocol for the extraction of anthocyanins from plant material, the first step is the sample preparation. The sample is prepared by converting the plant material into a powdered form after mixing it with liquid nitrogen. Is it necessary to mix the plant material with liquid nitrogen or can we proceed without liquid nitrogen to convert plant material into powdered form?
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The liquid N2 is to keep it cold, make it brittle, and exclude O2. The success otherwise depends on the properties of the particular material.
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Anthocyanins are rich in antioxidant properties. Previous research has shown that red rice contains a good amount of anthocyanins. While there are a number of laboratory procedures for anthocyanin extraction, is there a standard operating procedure(SOP) for its extraction from red rice to be used in the industry? Any suggestions are welcome.
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Concerned researchers can describe the topic.
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How to estimate anthocyanin content of vegetable sample from OD value in spectrophotometry method?
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During aging or storage, the concentration of monomeric anthocyanins decreases rapidly, while low level of polymeric anthocyanins generates, and the color of wine turns brick-red gradually. In my opinion, the contribution of polymeric anthocyanins to the old wine color is relatively small due to its small amounts. So, where do the brick-red or brown color come from? Why is it so difficult to detect these pigments through HPLC-MS? Is there any method to measure them?
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I’m wondering if the oxidation of condensed tannin and the auto-polymerized flavanols also result in the brown color. And those compounds’ structures were well studied in the model solution during the past decades.
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Hello, We are looking for the information and references related to the normal ranges for Anthocyanin & anti-nutritional (Phytic acid, tannin & Gliadin to gluten ratio) factors in foods/plant leaf/fruits. We found many references related to anti-nutritional (Phytic acid, tannin & Gliadin to gluten ratio) factors but no one mentioned about the normal ranges which are suitable for consumption. Hence, we are looking for this information. Regards, Raghavendra HL
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Please look at the following below links which may help you in your analysis:
Thanks
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Dear Researchers,
I did Transcriptome of my two Fruit varieties at developmental, ripening and harvesting stage for the comparison of their differential gene expression profile, such as Sugar acids, Anthocyanins, Color, Volatile, Fatty acids, etc
Now the company gave me results in different files format and it's little complicated for understanding and analysis, so if you have experience and have some websites and software for transcriptome data understanding and analysis then please share with me
Thanks a lot
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omics share website?
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I use 0.5 g berry extract which i dissolve in 50 µl water and then i add 100 µl 3 sulfuric acid to make the extraction more acetic. After this point i add 1 ml 80% methanol to do the extraction, spin down, and transfer 1 ml supernantant to another tube. Then I add another 1 ml methanol to the pellet and repeat the procedure so I now have 2 ml extraction.
In the assay the methanol hopefully containing my anthocyanin is diluted 1:40.
The formula look like this, but i am unsure how to calculate the amount of anthocyanins in the dried extract (W/W). Will i then say i have 0.05 g in 2 ml methanol as i would expect the anthocyanins to be in this fase ex (0.05g/0.002 L)*40?
The formula looks like this:
Anthocyanin concentration=(A*MW*DF*1000)/(ɛ*1)
I really hope somebody will be able to help
Thanks,
Christine
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I agreed with Duried answer he mentioned an easy protocol to determine the anthocyanins conc.
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when determining the total anthocyanin content using a SO2 bleaching protocol, the results are showing only the quantity of anthocyanins (not bonded to tannins)? And the same question for the tannins determined by absorbance measurement at 550 nm after the acid hydrolysis. Will I get results of tannins not bound to anthocyanins? Or can I speculate that some molecules will be bounded? Thanks!!
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Anthocyanins (At) are present in different forms: free anthocyanins (Al) and anthocyanins combined with tannins (Ac), some of which are bleached by SO2 (TA), while the rest is unaffected (TAT). In conclusion, by bleaching with SO2 some of the anthocyanins bonded to tannins (TA) will respond.
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There are many color of anthocyanin pigments?
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Dear scientists,
I have been using the method of pH differentiation to measure anthocyanins. However, I wanted to know how to differentiate between polymeric, monomeric and copigmented anthocyanins. I have seen the following methods in and :
Polymeric anthocyanins: A520 with SO2
Monomeric anthocyanins: A520 -A520 with SO2
Copigmented anthocyanins: A520 with acetaldehyde - A520
Total anthocyanins: A520 with acetaldehyde
But I have some questions regarding the basis for these assays:
  • Can SO2 affect only monomeric anthocyanins because the polymeric ones don't have available heterocycle ring for the molecule? The spatial hindrance?
  • Can it be assumed that the formation of a long chain of amino anthocyanins will cause an increase in the absorbance value?
  • How the copigmented anthocyanins relate to the total amount of the anthocyanins - how that affect the absorbance, what is the relation between copigmented anthocyanins and acetaldehyde?
  • Do the measurement with acetaldehyde relate to the pH differentiation method? Why doesn't the latter use acetaldehyde?
Thank you in advance for your collaboration.
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goods quistion
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like anthocyanin and flavonoids pigments
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Proteins and carbohydrates are known as primary metabolites, as they are essential for the vital development of plants, while secondary metabolites (such as anthocyanins, flavonoids, etc.) appear from the adaptation of plants to their environment (attract pollinators, repel pests ...) but not for growth and development. They are related in that some secondary metabolites are synthesized by metabolism of the primary metabolites.
Regards
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In a dye-sensitized solar cell using TiO2 as the semiconductor, FTO or ITO glass, KNO3 as the electrolyte and clorophyll and/or anthocyanin as the dye, why is the resistance observed on the cell so high? How can I low it down?
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Dear Gabriela,
I may come late. Hope you are already solved the problem. The resistance at the terminal of the solar cell is due to either the bulk resistances of the consisting layers of the solar cells or the resistance of the interfaces between the layers.
According to your solar cell description, the die is the layer that may have the highest resistivity. So, it must be very thin in the order of 20 nanometer. Also there are are two dominant interfaces the interface between the Die and TiO2 as well as the Die and electrolyte. These interfaces must be verified for the energy band matching.
If you display the the dark and the illuminated I-V characteristics one can make a better diagnosis of the solar cell structure.
Best wishes
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Based on literature that Pb is suitable with Dhitiazone as regaent in base medium but in the other side I'm going to use anthocyanin to chelate Pb. as we know that anthocyanin is in the optimum with pH 4. Am I supposed to change the reagent with acid one or I dont need to use Dhitiazone when I already use anthocyanin? (It means Dhitiazone is used in blank solution only)
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Some compounds of Pb turn blue when carotene is added.
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mixed sample of anthocyanins as a sensitizer for DSSC
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Dear Babangida,
welcome,
i can give give a conceptual answer. As these
anthocyanins acts as the Dies for the solar cells they can be mixed together with the proper ration and used as an active material for absorbing light and affecting photogenartion. By mixing you may get a die with more absorption from the incident solar radiation and so increase the photo current.
From the practical point of view you can run such experiment and evaluate it. You use every die separately and then you can combine them and assess your results.
Wish you success
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I need the full protocol for analysis Chlorophyll a and b, and Anthocyanin for hydroponics lettuce and the ideal solvent for analysis such as acetone for cholophyll and methanol for anthocyanin.
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I am wondering what the expected zeta potential (ZP) of anthocyanins in an aqueous solution at pH 3 compared to pH 5.
My thinking is that ZP at pH 3 would be more positive (higher absolute value) than at pH 5. For instance, ZP at pH 3 is +10 but at pH 5 would be +1. I don't know if these numbers are the actual truth.
I expect ZP to be positive because of the overall positive charge of the flavylium cations dominant at pH 3. The structural equilibrium of anthocyanins shifts to more neutral structures as pH increased thus ZP would decrease (be closer to zero).
I would appreciate help thinking through this question and any sources that may discuss the ZP of anthocyanins.
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The presence of a hydrocolloid is important to understanding what might be happening.
If you measure zeta potential via light scattering (Zetasizer, ZetaPALS etc) then it requires substance(s) that scatter the laser light. A true solution won't do this but a colloid will. As Leo Vleugels explained, zeta potential is a concept related to colloids and not solutions.
What is your hydrocolloid? Is it at a sufficient concentration to form a gel?
If the hydrocolloid is negatively charged then decreasing the pH could neutralize it and cause precipitation. It could be that the anthocyanin just happens to be there and is acting as an indicator of the change in the nature of the hydrocolloid!
What you would need to do is measure the "zeta potential" of just the hydrocolloid at different pHs. I put the term in quotes because a gel won't move in the same way as a dispersed particle but the instrument may still detect vibration due to the electric field. Typically, this would require using the so-called phase analysis mode (also known as fast field reversal and monomodal). Not seeing a difference will not rule out the mechanism but a difference will support it.
In addition, you can simply observe the visual appearance of the hydrocolloid as a function of pH. Shine a laser pointer through the sample and see if there is an obvious change. This will help determine if it is simply the hydrocolloid changing its physical properties.
Another possibility is that the anthocyanins may become protonated at low pH leading to a positively-charged polyions. These may act as electrolytes that - if at sufficient concentration - could destabilize the hydrocolloid. What is the molar concentration of the anthocyanin? I would guess that if it is above a few tens of millimolar then this could happen.
I don't known enough (okay - anything) about the solution behavior of anthocyanins. My suggestions above are based on general concepts and may be irrelevant in your case.
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I will isolate cyanidin compounds from local fruits. we try to get pure cyanidin which is free from the 3-glucoside chain. if you have the right and effective method to get this compound, please share with us.
Thank you
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In my experience, anthocyanidins should be concentrated/separated by using Sephadex LH-20. A well-designed method for preparative RP-HPLC will be helpful for the isolation of compounds. I suggest you contact Dr. Jorge E. Wong-Paz , he is experienced in the purification of procyanidins.
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I have a methanol extract of 3-deoxyanthocyanins (not water-soluble) from sorghum and I need to remove the alcohol-soluble proteins from this extract. What would be the best way to do this?
Solubility is reduced with lower temperatures and I do see precipitation of protein when I freeze the extracts (-20C).
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Using some precipitation agent that selectively reacts with the proteins, but not with the 3DAX may be one way.
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I'm looking for a simple method to concentrate anthocyanin extract and to me precipitation seems the easiest. Any suggestions would be appreciated.
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I will isolate the anthocyanin compounds from the callus. I try to get anthocyanins. Is it better to estimate anthocyanins in dry callus or fresh callus?. If you have an idea or information, please join us.
Thank you
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Anthocyanin compounds are extracted from the plant raw material with acidic alcohol.
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I want to research on anthocyanins however I'm not finding what I want. I want to do research by isolating and then seeing how different ph and temperature change the pigment. I just can't seem to find what everyone else has researched and I don't want to duplicate results if I can do something new.
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Look up articles by R. Brouillard, like
DOI: 10.1021/ja00447a012
or
DOI: 10.1016/S0040-4039(00)84978-3
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i tried to inject the sample without any pretreatement as did for anthocyanins unfortunately in this case i couldnt obtain any pic.
the using analysis method is HPLC-DAD-MS
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I just want to know which solvent system and spraying reagent will be used for TLC of anthocyanine.
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If you are looking anti-oxidants this will work: 0.05% (m/v) DPPH methanolic solution. After immersion, remove excess DPPH, plates keep in a dark box for 12 h and then photographed under visible light
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Why doesn't the anthocyanins/flavonoids accumulation in petals result in a colored phenotype?
Carotenoids cannot accumulate in the petals because Arabidopsis lack chromoplasts, essential for accumulation or storage of carotenoids. But what is the reason behind flowers not having pigmentation from flavonoids?
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By using a transcription factor from blood orange we were able to induce the appearance pink petals in some of our transgenics, thought the most robust and consistent phenotype was anthocyanin accumulation in mature embryos. I think this supports your above stated conclusions.
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I'm looking for TLC solvent system which suitable with anthocyanin
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you may consult this book " Phytochemical Methods A Guide to Modern Techniques of Plant Analysis, page 69" where you can find different systems to investigate different types of anthocyanin.
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relationship between chlorophyll and anthocyanins content
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Thank you so much for your kindly help.
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Hey folks ! I am doing something new experiments and each time i am getting the production of anthocyanins in my Arabidopsis seedlings. is it any ways to quantify them and also the modifications of them by glucosylation/glycosylation?
If you guys have any idea kindly send me the protocols, that would be great !
thank you in advance!
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Collaborate with experts in mass-spectrometry, i.e., LC-MS/MS and then you would get good 'specificity' identification and better quantification than colorimetric/ spectrophotometric (not sensitive enough) and enzymatic assays. Consult with experts on pigments as well.
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How does one make anthocyanins free of hygroscopicity without excipients and freezing?
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thank you very much rodrigo castaneda
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Hi everyone,
Is Horwitz equation suitable for the validation of anthocyanins extraction and analysis method?
Thank you
Best regards
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Hello, Horwitz equation has some limitation but you can use if you have all sufficient data. But I think if you have all data for Horwitz so the better way is to perform validation with the use of classical analytical parameters which is more easy and reliable way. Limitation connected with the use of Horwitz equation is attached.
Hope this info will help you !
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The signals shown in the attached file, lower panel, appeared as more or less constant background signals in LCMS analyses (ESI, negative mode, qTOF). Conditions and materials have been routinely used with that instrument without problems: binary gradient of water and acetonitrile, each with 0.1% formic acid, at 0.4 mL/min over a Dionex Acclaim Polar Advantage II 2.1x 100 mm column. Steel tubing in front of and PEEK tubing behind the column. The contaminant does not appear in positive mode. As we measure in positive mode most of the time it is not possible to precisely find out when the contamination took place. The first analyses that have shown the signals were acid hydrolysates of plant polyphenols (proanthocyanidins) which contained 4 M HCl. 5 µL were injected of six samples (first in positive, then in negative mode).
I have tried to learn more about that contaminant by trying some simulations of the isotope pattern (upper panel). The pattern at lower m/z values (m/z 160.8) matches 3 chlorine atoms, the other one 4 chlorine atoms, and the difference between their supposed monoisotopic signals at m/z 160.8 and 195.8 is 35.9698 u (HCl). The simulations are for Si2Cl3 (m/z 160.8) and Si2Cl4 (m/z 195.8), which was the best match so far but the m/z deviation of about 0.03 is far to big and the isotope pattern does neither fit well.
Maybe someone is familiar with this contamination or has some idea about it.
Greetings and sincere thanks in advance,
Jandirk Sendker
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Guess what, Jandirk, your spectrum in negative mode matches Fe(II)Cl3 and Fe(III)Cl4 near perfectly. I am willing to bet 2 cents on that.
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I'm wanting to find a dietary questionnaire that allows me to evaluate the quantity of phytochemicals in the human diet, preferably validated. It would be useful if the results are broken down in categories such as flavonoids, isothocyanates, anthocyanins etc. Any help much appreicated.
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Not just food specialists
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In humans will the sugar moiety be metabolised after cleavage of anthocyanins?
Normally the aglycone is absorbed from the intestine, does the same happen to the sugar moiety?
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Hello Terun Desai,
I am sorry, I only realize to-day you asked this question.
The answer of Paulo is quite clear and does not need further comment. Hydrolysis of anthocyanins gives back anthocyanidins and the free sugar moiety.
However, I am just willing to tell you that almost all those natural heterosides (including anthocyanins) are of "bêta"-configuration, and, we (animals) have no enzymes that could hydrolyze such heterosidic bounds. Only gut microflora do possess the necessary bêta-glycosidases, and can metabolize such bêta-O-heterosides. Therefore, to my opinion, the sugar moiety has almost no chance to be metabolized by humans (after resorption). It will be rather used by gut microflora (thus, metabolized, as Paulo told you).
By the way, what about the "freed" anthocyanidins: it still is a cation (quite acidic compound) whom resorption by the colon is not obvious at all ... Would you have any information on its fate?
At the same time, because of the higher chemical instability caused by the formation of the free OH group at the C-3 position. the pyrilium ring could easily degrade: opening leads to alpha dicarbonyl derivatives, that further break down in 2 parts! We have shown earlier that human flora implanted into gnotoxenic mice could metabolize such "flavonoids" into acid-phenols, phenyl-lactones ... which were found into the blood (resorption):
C. Brezillon, S. Rabot, C. Philippe, J. Durao, C. Cheze, J. Vercauteren, in Polyphenols Communications 98, XIXth International Conference on Polyphenols, Lille (France), 1–4 sept. 1998, F. Charbonnier, J.-M. Delacotte, C. Rolando, Eds., Lille, 1998, vol. 1, p 11–12).
Philippe, C., S. Rabot, C. Brézillon, J. Durao, C. Chèze and J. Vercauteren, Excretion pattern of dietary catechin in gnotobiotic rats associated with a human intestinal microflora, in Polyphenols Communications 98, F. Charbonnier, J.-M. Delacotte, and C. Rolando, Editors. 1998: Lille. p. 67-68.
Deprez, S., C. Brezillon, S. Rabot, C. Philippe, I. Mila, C. Lapierre and A. Scalbert, Polymeric proanthocyanidins are catabolized by human colonic microflora into low-molecular-weight phenolic acids. J Nutr, 2000. 130(11): p. 2733-8.
Good luck for your research.
Joseph Vercauteren
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I would need a semi-noncomplex protocol for an anti-alpha glucosidase assays in anthocyanins
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Hello dear,
You can evaluate in vitro alpha-glucosidase inhibition assay according to method of Matsui et al., 2007.
Materials and reagents for assay-
1. Alpha-glucosidase enzyme.
2. Substrate: Sucrose.
3. Positive control: Acarbose.
4. Sodium dihydrogen orthophosphate (NaH2PO4.2H2O).
5. Disodium hydrogen phosphate (Na2HPO4.2H2O).
6. Glucose reagent.
7. Working solution (Phosphate Buffer; 80 mM, pH 7.0, 25 oC)
Best of Luck
Dharmendra Arya
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Hello,
Can someone suggest me a good spectrophotometer for anthocyanins/polyphenol/antioxidant properties quantitation?
I am followig the AOAC Official Method 2005.02.
Are there any specification on this instrument that I have to take in consideration?
Thank you
Arianna
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Dear Arianna,
Any good spectrophotometer can do the job. But I would like to give you a suggestion. Go for HPLC instead of spectrophotometric estimation. I had done a huge work on quantification of secondary metabolites, but couldnt get published in any good journal. All editors asked me to come up with HPLC data as spectro is backdated.
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I measure anthocyanin content in my samples. I measured the absorbance and put the values in equation. I am confused about the unit of of concentration. it should be mg/L?
the initial concentration of my ample was 2g/20mL so how can I calculate the concentration of anthocyanins in initial extract (2g/20mL)?
Please help me
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Dear S.Z
Before changing the units, firstly we must know what kind of sample, secondly the used method. I can help you because i am working in anthocyanins in fruits...
Best regards
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tomatos can be genetically modified ,so that they produce anthocyanin which can reduce cancer?
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Reducing cancer is not at all identical to preventing cancer. Rephrase your question!
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Anthocyanins are water-soluble and can easily degrade due to its environment . Any recommendations?
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Thank you!
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I want to calculate anthocyanins contents through ph differential method. i calculate for different quantities like 10g, 5g, etc. Now i want to calculate is their any significant difference in anthocyanins contents in different doses. which test I will apply?
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Dear Sir. Concerning your issue about the determination of anthocyanins contents in different doses of same sample. Initially 1g of skin and 25 mL of water were used in triplicate, protected from light. The filtrate absorbance readings were taken at l 535nm, in a Micronal B 382 spectrophotometer. In the anthocyanin concentration calculations were taken into account the dilution factor and the cyaniding 3-galactoside coefficient of extinction (98.2) (Fuleki & Francis (1968); Lees and Francis (1972).
Total anthocyanins (mg cyanidin g-1) = Absorbance x dilution factor / 98.2
After established the best extraction methodology, other solvents were evaluated: ethanol 95% P.A., aqueous ethanol solution 50%, methanol 50% and acetone 30%, using the volume of chosen extractor (15 or 25 mL). I think the following below links may help you in your analysis:
Thanks

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If my sample already has red color, can the result for phenolics using Folin's reagent at all be dependent on?
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max absorbance for anthocyans at acidic pHs are around 550 nm and FC is read at > 700 nm: red whould not hinder detection in the FC wavelength if not 2 complications:
- at the highly alkaline pH in the FC reaction anthocyanins will not stay coloured or at least not long. I'd be surprised if the blank stayed red. However it does go through a blue phase.
- And yes, anthocyans will react in the totel phenolic assay, they are polyphenols tooµ.
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we always find more anthocyanin in arabidopsis when grown on high sucrose gelling. how does sucrose induce anthocyanin? what is the mechanism underlying of that? it is a sterile and nutrient-aboundant condition in which plants do not starve or be infected? why does anthocyanin metabolited?
thank you so much for your expert intellgence support
huayi
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this review helps alto about anthocyanins
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Dear all. How long an anthocyanin rich-extract aqueous solution can be stored at -20 ºC, protected from light? Thanks in advance.
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refer this review article which covers all the details about anthocyanins...
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I want to quantify anthocyanin concentration in my samples using the spectrophotometer. Is there and ideal sample volume to use in the 96 wells plate?
Also, can pigment concentration data be compared if the volume between spectrophotometer samples varied (between different measurements)?
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Also, do not forget to implement the pathway correction option (usually available on 96-plate spectrophotometer)
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Can someone explain in simple terms how recovery of polyphenols in urine informs bioavailability of that compound?
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Oral or dietary polyphenols that appear in the Urine, after ADME (absorption, distribution, metabolism and excretion) are assumed to be bioavailable (BV). In a agreement with Professor Shi, metabolims of polyphenols could lead to "underestimation" of the bioavailability. So "urinary bioavialablity" is the lower estimate whist the actual could be grater? In principle, to improve the accuracy of BV determinations the excreted (Urinary) value should combine the net concentration of all metabolites and non-metabolized polyphenols (not easy to measure).
Alternative measures of bioavailabilty exist as you know.
Pharmacolgists define BV as; = Area under curve -with oral route/ Area under curve IV route.
Nutrition scientists consider "digestbility" or Retention as more relevant or practical. Nutrient digestbility = 100*(Intake - (Feceal + Urinary)/ Intake.
I have not thought much about this but, I think the general concensus is that all current definitions and measures of BV have some limitations (?). It would be interesting to see what others think.
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why does some plants accumulate anthocyanin, and others accumulate carotenoids ?
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see the followings
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Betalains and anthocyanins are said to be mutually exclusive in a plant species (Stafford HA, 1994, Plant Sci. 101:91-98). However, Harris et al. (2012, BMC Plant Bio 12:34) reported that betalains can be induced in anthocyanin-producing plants through genetic engineering and substrate feeding, possibly because of a background enzyme. 
1. Alternatively, can betalains be induced in plants that normally produce anthocyanins in an in vitro system using plant growth regulators?
2. Would spectrophotometric analysis be sufficient to detect the presence of both pigments in any given sample?
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Dear Emmanuel
In a study to establish callus production in Alternanthera sessilis grown under different combinations of growth regulators and light qualities and to assess whether these factors can increase betalain and flavonoid production, the most suitable treatment for callus formation and subsequent betalain and flavonoid induction was to combine a medium containing 6.7 μmol L⁻¹ 2,4-D and 9.0 μmol L⁻¹ BAP and blue light.
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Can anyone suggest me the easiest technique to detect anthocyanin derivatives/metabolites in rat urine? Should I collect the urine within 1 hour after the consumption? is it still possible to detect the anthocyanin derivatives after the rats were fasted for 24 hour (currently i do not have the knowledge of anthocyanin half life in sprague dawley rats)?