Christine Crawford

Synthetic #2 Formal Final Report

Synthesis of Piperonylonitrile
TA: Yi-Chun Lin (Jim) Chem 213, Section 2 April 11, 2013

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Introduction The ability to synthesize nitriles in different compounds is a very critical tool within organic synthesis reactions. Nitriles are organic substances that contain the cyano (C=N) group and are functionally known by virtue of their short, polarized triple bond.1 These cyano groups are widely found as starting materials and intermediates in organic synthesis.2 Because these groups can also be found so widely as starting materials or intermediates, nitriles have a wide commercial application in use for solvents, synthetic intermediates, pharmaceuticals, and monomers.1. The role of nitriles in medicinal agents has been steadily emerging as the number of nitrile-containing pharmaceuticals has increased over the past couple years. Currently, over 30 nitrile-containing pharmaceuticals are available and prescribed for a diverse variety of medicinal indications, and there are even more than 20 additional nitrile-containing leads in clinical development at this time.3 These drugs are also extremely effective because the nitrile group is quite robust and is usually not readily metabolized in the body.3 This means that the nitrile substituent in most of these nitrile-containing pharmaceuticals can pass through the body unchanged and be excreted without any extremely unwanted side effects. Nitriles are not only important for use in the pharmaceutical industry, but they can even been adapted to microscale work within an educational setting. Specifically, the synthesis of piperonylonitrile from piperonal allows students to give excellent yields of the corresponding nitrile in one easy 15-minute step, and is therefore an extremely helpful reaction in order for organic chemistry students to learn about the improvement in yields achievable by designing a synthesis that follows the shortest possible route.4 The synthesis of piperonylonitrile from piperonal and hydroxylamine hydrochloride under acidic conditions is shown in Scheme 1.

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- Hand drawn mechanism of Synthesis of Piperonylonitrile -

Scheme 1. Synthesis of Piperonylonitrile.

In the first step, the lone pair of electrons on the nitrogen of the hydroxylamine attacks the carbonyl component of piperonal and makes a tetrahedral intermediate with a hydroxylamine cation and an oxygen anion. This anion then attacks hydrochloric acid to form an alcohol. The leftover chloride ion then attacks one of the hydrogens of the hydroxylamine cation for reform hydrochloric acid and produces a hemiaminal. The alcohol substituent attached to the carbon will then again attack the hydrochloric acid, forming an oxygen cation (R-O+H2) and a chloride ion. The lone pair of electrons from the hydroxylamine will then attack the adjacent carbon to form a double bond between the carbon and nitrogen, releasing water as a leaving group and forming
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another nitrogen cation. The chloride ion will then attack the final hydrogen attached to the nitrogen to form the oxime intermediate. In a final step, the oxygen of the oxime will attack the hydrochloric acid to form another oxygen cation (R-O+H2) and a chloride ion. The chloride ion will then attack the hydrogen attached to the carbon of the double bond, therefore pushing these electrons to form a CN triple bond and expelling water as a leaving group. The synthesis of nitrile substituents is extremely important for synthetic organic chemistry, especially for the large scale production of medical agents and pharmaceuticals, as well as on a smaller scale use for the instruction of organic chemistry students. The purpose of this lab is to learn the reaction mechanism of the synthesis of nitrile groups from an aldehyde starting material on a small scale basis and quantify its efficiency of producing the product of piperonylonitrile from the starting materials. The product can be isolated from the reaction mixture and then analyzed by IR, 1H NMR, and this efficiency.
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C NMR instrumentation in order to quantify

Experimental Piperonal (200mg, 1.33mmol) and DMF (1mL) are added to a 5mL long-necked round bottom flask with attached air condenser and are stirred and heated to a boil (~170C). Hydroxylamine hydrochloride (120mg, 1.73mmol) is dissolved in DMF (1mL) and added drop wise to the boiling piperonal solution over a 2 minute period. Solution is then stirred and refluxed for 90 minutes, as it will gradually change from a colorless solution to a dark golden brown solution. Upon completion of the reaction, the solution is cooled to room temperature then placed in to a 50mL Erlenmeyer flask where it will sit in an ice bath for 5 minutes. Solution is then diluted with 10mL distilled water and allowed to crystalize over a two day period in the

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refrigerator. Crystals are then collected by vacuum filtration using a Hirsch funnel and washed with 1mL of cold water. The slightly wet crystals are allowed to dry until the next lab period (132mg, 64.7%) mp 90.5-93.0C; 1H NMR (400 MHz, CDCl3) (ppm) 7.19 (d, 1H), 7.01 (s, 1H), 6.84 (d, 1H), 6.08 (s, 2H); 13C NMR (400 MHz, CDCl3) (ppm) 152, 149, 129, 119.5, 111, 109.5, 106, 102 ;IR (ATR) (cm-1) 2919, 2219, 1606, 1482, 1254, 1026.

Results and Discussion As a class, nitriles are characterized by the presence of the CN (CN) group.5 Nitriles can be formed in many different ways, including simultaneous replacement of all three hydrogen atoms of a methyl group by nitrogen, by replacement of both the carbonyl and alcohol portions of a carboxylic acid group, or by the reaction of an aldehyde with a hydroxylamine.5 More specifically, reactions of aldehydes with hydroxylamines produce a nitrile substituent through the use on an oxime intermediate. The synthesis of piperonylonitrile is characterized by the exchange of the aldehyde of the piperonal reactant to form a nitrile substituent using hydroxylamine hydrochloride within an acidic reaction mixture. As the reaction takes place, the reaction mixture should change from a clear colored solution to a dark yellow/golden brown color. However, the piperonylonitrile does not precipitate or crystalize out of the solution during the reaction. Instead, the reaction mixture must be cooled within an ice bath for 5 minutes and then diluted with 10-15mL of distilled water. This distilled water helps to achieve a better crystallization because the piperonal starting material is more likely to stay dissolved within the water as the piperonylonitrile product is crystalized. To allow for an extremely efficient and complete crystallization of the piperonylonitrile product, the reaction mixture can be left in a refrigerator overnight, or even

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multiple days. The final crystalized product can then be vacuum filtered to isolate the piperonylonitrile product, which is a brown/grey colored crystal that resembles the looks of sheep wool. The product was analyzed by IR spectra data. The most notable change that occurs during this reaction is the substitution of the aldehyde on piperonal with a nitrile, producing an oxime as the intermediate product. An aldehyde of piperonal is characterized by the carbonyl C=O bond, which shows stretching peaks around 1650-1800cm-1. As can be seen from the spectral data (Figure 1, Supplemental Information), the isolated product from this experiment showed two peaks close to or within this range: one peak at 1852cm-1 and the second peak at 1730cm-1. These peaks are very small, but can support the conclusion that there is potentially a small amount of starting material still left over in the crystalized product. However, when converting piperonal to piperonylonitrile, the synthesis goes through an oxime intermediate. This intermediate would have a most notable stretching frequency around 3200-3600cm-1 which corresponds to the O-H bond attached to the nitrogen of the oxime. The IR spectral data of the final product of this synthesis does not show any peaks within this range that could support the conclusion that there is any intermediate left over within the crystalized product. Instead, only starting materials can be found in the piperonylonitrile product. Finally, the nitrile group of the piperonylonitrile product has a characteristic CN triple bond, having a stretching frequency between 2230 2250cm-1. As can be seen from the spectral data of the crystalized product (Figure 1, Supplemental Information), there is a very strong peak that can be found at 2219cm-1. Although this peak does not reside in the normal location for a CN stretching frequency, it is certainly very close and it can be concluded that this peak corresponds to the nitrile group.

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There are other strong peaks that are present within the IR spectra of piperonylonitrile (Figure 1, Supplemental Information), however, these peaks do not help to identify the purity of the sample as much because they are peaks that could be found for all three compounds: piperonal, the oxime intermediate, and piperonylonitrile. Some of these peaks include the peak at 2919cm-1 that corresponds to the stretching frequency of a C-H (sp3) bond, found on the one carbon of the 5-membered ring found between the two oxygens, as well as the C-O stretching frequency found at 1026cm-1, corresponding to the C-O bonds of the 5-membered ring. NMR was also a useful technique for characterizing the identity of piperonylonitrile as our crystalized product. Using a 60 MHz 1H NMR, spectral data (Figure 2, Supplemental Information) was indistinguishable as five peaks were all grouped very closely together, but only had an integral value of 3 hydrogens. To distinguish this data, 400MHz 1H NMR was used to produce a total of four peaks (Figure 3, Supplemental Information). The first three peaks of this spectra correspond to the hydrogens attached to the benzene ring of piperonylonitrile. The first peak is the appearance of 1 proton at 7.19ppm, which splits into a doublet. This one hydrogen can be associated with the C-H bond of the benzene ring closest to the nitrile group and having a single hydrogen on the adjacent carbon. Normal C-H bonds of benzene rings are found between 6.5-8.0ppm. The reason for this hydrogen to be found at the higher end of this spectral range is from the deshielding effects of the nitrile group attached to the benzene on the adjacent carbon. The other hydrogen located next to the nitrile group is located at 7.01ppm and forms a singlet peak. This peak is a singlet because there are no other hydrogens on adjacent carbons next to this C-H bond. However, the peak is not at as high of a ppm value as the first hydrogen because although there are deshielding effects from the adjacent nitrile group, there are also shield effects from the adjacent ether group of the 5-membered ring. Finally, the last of the hydrogens from the

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benzene ring can be associated with the peak at 6.84ppm. This peak again is a doublet because of the hydrogen on the adjacent carbon, but it is again lesser in ppm value because of the shielding effects of the ether group attached at the other adjacent carbon of the 5-membered ring. The final peak found on the 400 MHz 1H NMR is a singlet peak located at 6.08ppm, and has an integral value of almost 2. This last peak is associated with the two hydrogens attached to the carbon of the 5-membered ring, in between the two oxygens of the ring. There are no other peaks found on the NMR spectra of the crystalized product, so from this data we can conclude that the crystals are extremely pure. If there were some of the piperonal starting material left over in this sample, there should have been a peak around 9.0-10.0ppm, corresponding to the hydrogen of the aldehyde substituent in piperonal. Also, if any of the oxime intermediate were present in the final crystals, there would be a broad peak visible somewhere between 0.5-5.0ppm corresponding to the O-H bond of the oxime. However, because this peak is also not present in the spectra, we can conclude that this intermediate is also not present in the final synthesized sample, and only piperonylonitrile has been isolated from the reaction. A further examination of the synthesized crystals by 400 MHz 13C NMR (Figure 4, Supplemental Data) can conclude the crystals are pure piperonylonitrile. The
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C NMR spectra

shows 9 different signals of carbon atoms. Piperonylonitrile contains only 8 carbon atoms, however the one extra carbon found in this spectra is due to the carbon of the chloroform-d (CDCl3) used as the solvent for the NMR tube. Discounting, this chloroform-d peak located at 78ppm, the 8 peaks found on the
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C NMR spectra are located at 152ppm, 149ppm, 129ppm,

119.5ppm, 111ppm, 109.5ppm, 106ppm, and 102ppm. The first two peaks at 152ppm and 149ppm fall within the range of carbons that are part of a benzene ring. These two specific carbons of the benzene are most likely the two carbons that are attached to the oxygens of the 5-

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membered ring as well. This is most likely the case because the extra attachment of these oxygen groups will tend to increase the displacement of the peaks along the spectra. The third peak, found at approximately 129ppm corresponds to the carbon of the benzene ring that is attached to the nitrile group. Nitriles are usually located between 110-125ppm on a 13C NMR spectra, and so a carbon of a benzene ring attached to this nitrile should be just slightly above the range of a nitrile. The nitrile carbon is then found at the next peak at about 119.5ppm. It is known that this carbon corresponds to the CN of the piperonylonitrile because it falls within the range of nitrile groups. After this, the next 3 peaks of the 13C NMR spectra (Figure 4, Supplemental Information) pertain to the three last carbons that are a part of the benzene ring. The first peak, located at 111ppm, most likely corresponds to the carbon of the benzene ring located in between the adjacent carbons with attachments of the nitrile group and the oxygen of the 5-membered ring. The next peak at 109.5ppm corresponds to the carbon located on the opposite side of the nitrile group. The last of the benzene carbons resides with a peak at 106ppm. Finally, the last carbon of piperonylonitrile found on the
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C NMR spectra is located at the tip of the 5-membered ring in

between to the two oxygen atoms, and shows a peak at about 102ppm. This peak is the lowest of the 8 carbons of piperonylonitrile because it is an R2CH2 group. Usually, these groups are found more towards the lower end of the NMR spectrum, but because the two R-groups attached to the carbon are ethers, the peak gets shifted up the spectra more towards the carbon-oxygen single bond ppm range. The IR, 1H NMR, and
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C NMR analysis spectra for the synthesized product have

confirmed the identity of piperonylonitrile as the pure crystals formed from this synthesis process, and show little to no signs of any possible impurities. Although there were signs of

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possible leftover starting materials found in the IR spectral data, there were no signs of this contamination from either the hydrogen or carbon NMRs, therefore it is possible to conclude that this reaction was carried out to produce a yield of a very pure product. The amount of piperonylonitrile crystallized and isolated during this experiment returned a 67.4% yield. Although this yield may be slightly lower than desired, it is still a significantly decent yield for the product in this reaction. This lower percent yield could be due to the fact that not all of the starting materials had actually reacted during the 90 minute time period, as seen from the trace amount of piperonal in the IR spectra, and so this would lead to some sources of error and instead would need to find the corrected percent yield of piperonylonitrile from only the amount of piperonal and hydroxylamine hydrochloride that reacted during the experiment. In order to further test the purity of the 67.4% yield of piperonylonitrile, melting point range determination was carried out. Melting point of the isolated product produced values of about 90.5-93C. The accepted melting temperature range for piperonylonitrile is between 9193C, so this observed melting temperature range is almost exactly on point with the accepted vales. The barely lower starting temperature of this range may be due to the trace amounts of piperonal found in the sample, but overall it can be concluded that the crystalized and isolated compound from this synthesis is a pure piperonylonitrile crystal. The synthesis of nitrile substituents is an extremely important aspect of organic chemistry and synthetic organic reactions. Nitriles, such as the piperonylonitrile synthesized in this lab, are useful for not only the large scale production of medical agents and pharmaceuticals, but for use in model reactions to teach organic chemistry students the mechanism of the synthesis of nitrile groups from an aldehyde starting material. This experiment was able to focus on the synthesis of piperonylonitrile and then analyze the product for both quantity and percent yield as well as the

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purity of the sample. The product was analyzed by IR, 1H NMR, and determine the identity and purity of the product.

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C NMR in order to

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Works Cited 1. DeVito , Stephen C. Chapter 10: Designing Safer Nitriles. In Designing Safer Chemicals. American Chemical Society: Washington D.C., United States, 1996; pp 194-223. http://pubs.acs.org/doi/pdf/10.1021/bk-1996-0640.ch010 (accessed 4/9/13)

2. Prasad, Shreenath; Bhalla, Tek Chand. Nitrile Hydratases (NHases): At the Interface of Academia and Industry. Biotechnology Advances. [Online] 2010, 28, 725-741. http://www.sciencedirect.com/science/article/pii/S0734975010000716 (accessed 4/9/13)

3. Fleming, Fraser F.; Yao, Lihua; Ravikumar, P. C.; Funk, Lee; and Shook, Brian C. Nitrile-Containing Pharmaceuticals: Efficacious Roles of the Nitrile Pharmacophore. J. Med. Chem. [Online] 2010, 53, 79027917. http://pubs.acs.org/doi/pdf/10.1021/jm100762r (accessed 4/9/13)

4. DeMott Jr , James M; Kelley, Charles J. An Alternative One-Step Procedure for the Conversion of Piperonal to Piperonylonitrile. J. Chem. Educ., 2001, 78, 780. http://pubs.acs.org/doi/pdfplus/10.1021/ed078p780 (accessed 4/9/13)

5. Nitriles. In Nomenclature of Organic Compounds: Principles and Practice. Fletcher, John H., Ed.; Dermer, Otis C., Ed.; Fox, Robert B., Ed. American Chemical Society: United States, 1973; pp241-245. http://pubs.acs.org/doi/pdf/10.1021/ba-19740126.ch031

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