An experiment between Isovanillin and Vanillin

Isovanillin is a phenolic aldehyde, an organic compound and isomer of vanillin.It is a selective inhibitor of aldehyde oxidase. It is not a substrate of that enzyme, and is metabolized by aldehyde dehydrogenase into isovanillic acid.

Vanillin is a phenolic aldehyde, which is an organic compound with the molecular formula C8H8O3. Its functional groups include aldehyde, ether, and phenol. It is the primary component of the extract of the vanilla bean. Synthetic vanillin, instead of natural vanilla extract, is sometimes used as a flavoring agent in foods, beverages, and pharmaceuticals.

There is an experiment about enzymatic oxidation of vanillin, isovanillin and protocatechuic aldehyde with freshly prepared Guinea pig liver slices. Let me show you the background, methods, results and conclusions of this experiment briefly.

Background: The oxidation of xenobiotic-derived aromatic aldehydes with freshly prepared liver slices has not been previously reported. The present investigation compares the relative contribution of aldehyde oxidase, xanthine oxidase and aldehyde dehydrogenase activities in the oxidation of vanillin, isovanillin and protocatechuic aldehyde with freshly prepared liver slices.

Methods: Vanillin, isovanillin or protocatechuic aldehyde was incubated with liver slices in the presence/absence of specific inhibitors of each enzyme, followed by HPLC.

Results: Vanillin was rapidly converted to vanillic acid. Vanillic acid formation was completely inhibited by isovanillin (aldehyde oxidase inhibitor), whereas disulfiram (aldehyde dehydrogenase inhibitor) inhibited acid formation by 16% and allopurinol (xanthine oxidase inhibitor) had no effect. Isovanillin was rapidly converted to isovanillic acid. The formation of isovanillic acid was not altered by allopurinol, but considerably inhibited by disulfiram. Protocatechuic aldehyde was converted to protocatechuic acid at a lower rate than that of vanillin or isovanillin. Allopurinol only slightly inhibited protocatechuic aldehyde oxidation, isovanillin had little effect, whereas disulfiram inhibited protocatechuic acid formation by 50%.

Conclusions: In freshly prepared liver slices, vanillin is rapidly oxidized by aldehyde oxidase with little contribution from xanthine oxidase or aldehyde dehydrogenase. Isovanillin is not a substrate for aldehyde oxidase and therefore it is metabolized to isovanillic acid predominantly by aldehyde dehydrogenase. All three enzymes contribute to the oxidation of protocatechuic aldehyde to its acid.

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Synbiotics probiotic supplement for children has been discovered containing vanillin

Recently, the synbiotics probiotic supplement for children has been discovered containing vanillin. The self-proclaimed "baby little sick, my mother worry less about" children's products quickly attracted the attention of many parents, vanillin is what? Would adversely affect children do?

Vanillin (CAS NO: 121-33-5) usually divided into methyl vanillin and ethyl vanillin. It is an edible spices, from the outside looks usually white or slightly yellow crystal, with vanilla aroma and rich milk, vanilla can be found in the seeds can also be synthetic.

Vanillin in the food industry is very widely used, can be used for the preparation of tea, chocolate, butter and so on. After years of experimental research, FAO and the World Health Organization (FAO / WHO) Joint Expert Committee established vanillin highest daily intake limit of 0-10 mg / kg body weight, while the European Food Safety Authority established maximum limits for the is 47 mg per day. And all that does not exceed the current limit under the premise of using vanillin, do not worry about security.

Some people may worry that, in case of overdose businesses add it? In fact, it is almost unfounded. Because, in the actual production under normal circumstances no one high-dose use of vanillin. Is the so-called extremes meet, just like many other flavors, like vanillin, used in small amounts will add flavor; but if used in excess, it is no longer the flavor, but the bitterness.

Since vanillin security can be reassuring, then why this "children's probiotic supplements" products will be investigated and dealt with it. In fact, it is primarily a violation of our laws.

According to China's current food additive standard (GB2760-2011), for 0-6 months baby infant formula does not add any food flavor, which naturally includes a stage of infant formula. However, for six months or older infants and infant formula foods, including diblock and triblock infant formula, it is allowed to use vanillin, ethyl vanillin and vanilla extract, which is three kinds of additives maximum allowable volume of 5 mg / 100 ml, 5 mg / 100 ml, and in accordance with production needs in moderation.

View synbiotics this food and found it prominently says children products, advertising language also suggests that the baby's food, then it includes infants 0-6 months. The product packaging is also clearly indicate the main raw material containing vanillin, but suitable for people in this column, but the product is marked "no."

In other words, this type of synbiotics children children's probiotic supplements do not apply to infants 0-6 months. In fact, if it is clearly written on the label on the label of this product is not suitable for infants 0-6 months, then there is no problem, you can normally sell.

Parents are more concerned or infants eat such products will not be harmful to it? In fact, this product although a violation of our regulations, but it does not mean that will have a direct impact on the child's body. Provisions of GB 0-6 months baby infant formula does not add vanillin, vanillin certainly not because infants have a direct physical harm. Infant formula should be only essential nutrients, vanillin no nutritional value, so there is no need to add. The present study found that the infant milk powder containing vanillin possible impact may affect primarily infants grow up right food choices, because the addition of vanillin more fragrant milk taste better, baby prefers to eat, long Great post may prefer sweet foods, their health is not adversely affected.

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History of Vanillin tablets

1. Basic information of Vanillin tablets

Vanillin is a phenolic aldehyde, which is an organic compound with the molecular formula C8H8O3(CAS NO: 121-33-5). Its functional groups include aldehyde, ether, and phenol. It is the primary component of the extract of the vanilla bean. Synthetic vanillin, instead of natural vanilla extract, is sometimes used as a flavoring agent in foods, beverages, and pharmaceuticals. But what the most important usage is treatment for epilepsy in medicine community. Vanillin is the main Component of Vanillin tablet which is high-performance for epilepsy.

Vanillin tablets are synthetic, not Traditional Chinese Medicine. Vanillin tablet is a kind of anti epileptic drugs. It is used for the treatment of epilepsy and other various types of epilepsy, vertigo, adhd and so on.

2. Usage and Dosage of Vanillin tablets

Oral, 3 times a day, a 0.1-0.2g (1/2 or 1) or as directed by a physician; children dose reduce.

3. History of Vanillin tablets

In 1965, China Medicine University (formerly the Nanjing College of Medicine) was first synthesized with good anti epilepsy effect of vanillin. Initiated the anti epilepsy era for antiepileptic. At the same time, it caused international epilepsy countries such as Poland, United States and other countries attention.

In 1979, vanillin for anti epileptic subjects is listed as key scientific research item of Health Department of Jiangsu Province, and organized several related units to work together, and after 17 years of efforts, finally completed the pharmacology, toxicology, pharmacokinetics, quality standard, production process and clinical validation studies.

In January, 1982, through the identification of the treatment of epilepsy, we confirmed that Vanillin tablet has good curative effect on epilepsy, and no obvious toxic side effects.

In October 1982, Jiangsu Province Health Department formally approved the production of vanillin.

During the next 20 years, the process of vanillin is continuous improvement. Vanillin was renamed vanillin tablets, is widely used in clinical treatment of seizures and encephalopathy. Vanillin tablets have been approved as the country medicine accurate H320258 by the National Food Administration in January 29th, 2003.

During the 5 years from 2000 to 2005, China Medicine University Pharmaceutical Co Ltd, increase the vanillin tablets of the promotion, tens of thousands of seizures, encephalopathy patients in rehabilitation! At the beginning of 2005, a plan, named "improve the life quality of patients with epilepsy" started quietly. After that, Vanillin tablets began to expand outwards at an alarming rate, a potential from provincial hospitals.

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Initial Assessment for Human exposure in Vanillin

Vanillin is a phenolic aldehyde, which is an organic compound with the molecular formula C8H8O3. Its functional groups include aldehyde, ether, and phenol. It is the primary component of the extract of the vanilla bean. Synthetic vanillin, instead of natural vanilla extract, is sometimes used as a flavoring agent in foods, beverages, and pharmaceuticals.The cas no is 121-33-5.

Vanillin occurs widely in plants in the nature, usually as a glycoside bound to sugar or as a precursor to vanillin bound to the large lignin molecule abundant in wood. Free vanillin in the environment will be distributed to the aqueous compartment, and there is no tendency to bioaccumulation. The emission of vanillin from the vanillin production and from consumer products to the environment, is not considered to represent any biohazard. In the previous article,we have talked about the question that how does human expoeure in vanillin. This passage,I will give the Initial Assessment.

In the human health risk assessment the consumer exposure (by oral intake) is taken as the ADI value(10.0 mg/kg/day).

Occupational exposure has been identified as inhalation of vanillin dust by operators in the packaging area of the factory. This exposure has been quantified to a maximum of 0.5 mg/kg/day. This level is likely to never be reached since the amount of vanillin in the total dust is estimated to only about 10 %. Additionally, much less than 1 % of the product has a particle size small enough to reach the lungs.

Skin exposure to vanillin has not been quantified, but both consumers and workers will be subject to aminor extent of suchexposure.

1.Assessment from acute and repeated dose toxicity

From the acute toxicity studies, the LD50 value for rats (3500 - 4000 mg/kg) gives a safety margin of 350-400 for consumers, and this is considered satisfactory. From the oral repeated dose toxicity studies a NOEL of 2500 mg/kg/day was observed after oral feeding to rats. This gives a safety margin of 250 for consumers use, and this is considered acceptable.

2.Assessment from reproduction and development studies

The potential toxicity of vanillin to the reproductive system has been studied in several connections. Microscopic and macroscopic histopathological evaluation of reproductive organs were performed in connection with repeated dose studies. In the mouse spot test and in a study to test the possible antimutagenic effect of vanillin, it was given to pregnant mice. The potential teratogenicity of vanillin has

also been tested in the developing chicken embryo test. There has been no sign of vanillin being toxic to the reproduction system, or to the developing embryo in any of these tests. Even though a reproduction/development study carried out according to OECD-guidelines is lacking, the present use and production of vanillin indicate no such risk.

3.Assessment from genotoxicity and cancer studies

The testing of the potential genotoxicity of vanillin is comprehensive. Mutagenicity testing in bacteria was negative. Testing in vitro in mammalian cells gave positive results in tests for sister chromatid exchange and chromosomal aberration in human lymphocytes. These results indicate that vanillin under certain testing conditions might be genotoxic. However, in vivo genotoxicity tests were negative. An overall evaluation of the test results indicate that vanillin is not likely to pose a genetic risk to humans.

The carcinogenicity studies, including a 2 years study in rats, did all give negative results. It is concludedthat the present use and production of vanillin represent little risk for genotoxicity and there are no signs of carcinogenicity.

4.Assessment from skin irritation and sensitisation testing

Skin irritation and sensitisation testing have shown that in some tests vanillin turned out to be a sensitiser and thus a potential allergen. These results are, however, not conclusive and the negative human datasupport the opinion that vanillin is not a human allergen.

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Human Exposure in vanillin

Vanillin is a phenolic aldehyde, which is an organic compound with the molecular formula C8H8O3. Its functional groups include aldehyde, ether, and phenol. It is the primary component of the extract of the vanilla bean. Synthetic vanillin, instead of natural vanilla extract, is sometimes used as a flavoring agent in foods, beverages, and pharmaceuticals.The cas no is 121-33-5.

Vanillin occurs widely in plants in the nature, usually as a glycoside bound to sugar or as a precursor to vanillin bound to the large lignin molecule abundant in wood. Free vanillin in the environment will be distributed to the aqueous compartment, and there is no tendency to bioaccumulation. The emission of vanillin from the vanillin production and from consumer products to the environment, is not considered to represent any biohazard.In this article,we will talk about the question that how does human expoeure in vanillin.

Firstly,let us talk about the Occupational exposure.

The industrial production of vanillin is a closed process. However, the milling of the dried crystals and the filling of the product into containers generate some dust to which a limited number of workers are exposed. Also some dust formation is expected in the food and beverage industry during addition of vanillin.

A study was carried out in the packaging section at Borregaard EuroVanillin, Norway, to assess the inhalation exposure of the operators to vanillin dust. The work in the packaging section is divided among 12-18 persons, meaning 2-3 persons per shift. On average, these workers are exposed to vanillin for 1 to 1.5 hours per day. For the rest of the day they conduct other operations where they are not exposed to

vanillin. The exposure was measured by a constant flow sampler (2 l air/min) carried by the operator during the exposure period. The total amount of dust collected corresponded to air concentrations of 4.2, 4.3 and 6.0 mg/m3 for the three consecutive days. From microscopic examination of the dust particles, only 10 % of the dust particles were considered to be vanillin.

Occupational exposure to vanillin other than from dust, is of little relevance. For the risk assessment, it is anticipated that occupational exposure is maximum 5 mg/m3. Assuming a person has a respiratory minute volume (RMV) of 20 l and an exposure for 6 hours per day at the limit concentration, will give a daily exposure by inhalation of 36 mg. For a 70 kg person this makes 0.5 mg/kg/day.

Absorption of vanillin by inhalation of dust will also depend on particle size. Sifting analysis of crystalline vanillin revealed that less than 1 % of the particles were smaller than 53 microns. Since only particles smaller than 5 microns are respirable, much less than 1 % of inhaled vanillin can be absorbed. This is valid for vanillin produced from both lignin and guaiacol.

Then,Consumer exposure.

The majority of industrially produced vanillin is ingested in the form of food and beverages. Minoramounts are applied topically as skin care products, perfumes etc. The global use of vanillin in food and beverages imply that almost every human globally is exposed to minute amounts of vanillin by ingestion.The individual doses and exposure can vary due to eating habits and preferences. An AcceptableDaily Intake (ADI) of 10 mg/kg has been agreed between FAO/WHO and EU. For a 70 kg person the ADI is 700 mg vanillin which as an example corresponds to minimum 700 g chocolate, or 7000 g of ice cream. For the risk assessment it is assumed that even persons with a high intake of vanillin containing food and beverages do not have a vanillin intake above the ADI.

Here,During the present reviewing of the available toxicity data for vanillin, no particular risk has been identified which should give reason to concern or additional toxicity testing in animals. The use of vanillin as a food additive is approved by authorities world wide, and FDA has granted GRAS status to its use. This is in agreement with the experience with vanillin in consumer products during many years without any confirmed report of adverse events.

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The aquatic effects of vanillin on the Environment

Vanillin is a phenolic aldehyde, which is an organic compound with the molecular formula C8H8O3. Its functional groups include aldehyde, ether, and phenol. It is the primary component of the extract of the vanilla bean. Synthetic vanillin, instead of natural vanilla extract, is sometimes used as a flavoring agent in foods, beverages, and pharmaceuticals.The cas no is 121-33-5.

Vanillin occurs widely in plants in the nature, usually as a glycoside bound to sugar or as a precursor to vanillin bound to the large lignin molecule abundant in wood. Free vanillin in the environment will be distributed to the aqueous compartment, and there is no tendency to bioaccumulation. The emission of vanillin from the vanillin production and from consumer products to the environment, is not considered to represent any biohazard.Now,this article will talk about the aquatic effects of vanillin on the Environment. I just talk about the effects for fish,Daphnia,Algae and Microorganisms.

1.Fish
The acute toxicity of vanillin to fish has been tested in Fathead Minnow (Pimephales promelas). The following LC50 values were observed for the following observation periods: 1 hr; 173 - 370 mg/l, 24 hr;100 - 131 mg/l, 48 hr; 68.3 - 123 mg/l, 72 hr; 57 - 123 mg/l and 96 hr; 57 - 123 mg/l. It was observed that fish stopped schooling, became hypoactive, swam at the surface and lost equilibrium prior todeath. There are no data available on the chronic toxicity to fish.
2.Daphnia
The acute toxicity to aquatic invertebrates has been tested in daphnia. After 24 hours exposure, the EC50 value was reported to be 180 mg/l. A 21 day reproduction study has been performed on daphnia, according to OECD 202 and GLP. After an exposure for 13 days to 100 mg/l, immobilisation occurred in all animals. The EC50 for immobilisation in the period 13 to 21 days exposure was estimated to 75 mg/l. The reproductive function was however more sensitive to vanillin, and after 21 days exposure the EC50 (reproduction) was 16 mg/l (24mg/l), NOEC 5.9 mg/l (10 mg/l) and LOEC 10 mg/l (18 mg/l) (nominal concentrations given in parentheses).
3.Algae
The toxicity of vanillin to algae was tested in a screening study in a variety of species. Only one concentration level (2 mg/l) was used. The green algae Scenedesmus obliquus and Chlorella variegata showed reduced growth after 3 days but no effect was observed after 7, 14 and 21 days exposure. When the diatoms Gomphonema parvulum and Nitzschia palea were tested for 2 mg/l of vanillin, the former showed no growth after 3 days exposure and reduced growth after 7, 14 and 21 days. The latter was unaffected after 3 days, but showed reduced growth after 7 days, however growth was normalised after 14 and 21 days. No effect was seen with the blue-green algae Cylinderospermum licheniforme and Microcystis aeruginosa. These results could indicate that green algae and diatoms are the most sensitive algae, but since this was a screening test using only one dose level, one should not attach too much importance to this finding. In another report, 50 % growth inhibition of Chlorella vulgaris was observed after 80 hr exposure to 152 mg/l. After 160 hr exposure to this concentration the growth inhibition was 30 %. No effects were observed at 15 mg/l and 1.5 mg/l.
4.Microorganisms
Another study tested the effect of vanillin on bacteria, yeast and blue-green algae in water. The effect of vanillin on anaerobic methane formation from sludge was measured, and 49 hr incubation with a concentration of 1,800 mg/l reduced the methane production with 50%. Photobacterium phosphoreum was however seen to be more sensitive to vanillin, and after 5 minutes incubation, an EC50 value of 58 mg/l was observed. Yeast (Saccharomyces cerevisiae) showed an EC50 value of 179 mg/l after 210 minutes incubation with vanillin.

To summarise the effects of vanillin to the aquatic environment, fish, bacteria and most algae seems moderately sensitive to vanillin. There are indications that some algae species are more sensitive to vanillin, and the reproductive function of daphnia was shown to be sensitive. A PNECaquatic based on the NOEC value (5.9 mg/l) found in the Daphnia reproduction test would give a value of 0.059 mg/l (onlyone long term NOEC, application factor=100). A PNEC value based on most sensitive EC50/LC50 value (LC50 fish = 57 mg/l) and an application factor of 1000 give a PNECaquatic of 0.057 mg/l. Photobacterium phosphorum being the most sensitive organism and using an application factor of 10 gives a PNECSTP of 5.8 mg/l.


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