pubs.acs.

org/jchemeduc Article

Toward Qualified Synthetic Chemist. Step 1: Building and Assessing

Basic Skills in Organic Synthesis
Dmitriy M. Volochnyuk,* Valeriya G. Makhankova, Alexander Yu. Lyapunov, Liliia S. Muravska,
Olexander M. Vitsenko, and Sergey V. Ryabukhin*
Cite This: J. Chem. Educ. 2023, 100, 3309−3319 Read Online

ACCESS Metrics & More Article Recommendations *

sı Supporting Information
See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.

ABSTRACT: An introductory experimental course to teach and assess

basic organic synthesis and identification skills has been designed. The first
Downloaded via Alexander Lyapunov on September 19, 2023 at 07:08:33 (UTC).

work of this course, which is based on the well-known hydrazone

formation reaction of benzaldehydes with Brady’s reagent, is presented.
The designed experiment has many advantages and perfectly matches the
goal of teaching rookie chemists. At the same time, the work allows one to
fasten such primary laboratory skills as a measurement of the mass of a
solid substance; measurement of the volume of a liquid; the use of a
laboratory apparatus for filtration under reduced pressure; making
calculations of yield neatness and accuracy in doing the work; the ability
of students to make and record qualitative observations. Apart from
teaching, the work emphasizes the quantitative assessment of students’
skills and the development of a clear and easy-to-use grading scale
including all basic quality parameters of a reaction product. Additionally, a
comprehensive analysis of how students’ mistakes contribute to the products’ yield and purity as well as the time required for the
accomplishment of the experiment was performed. The proper design of the experiment and precise quality control of the product
help to answer the question, “What have the students done wrong while performing the work?”.
KEYWORDS: High School/Introductory Chemistry, First-Year Undergraduate, General/Organic Chemistry, Assessment, Synthesis,
Identification

■ INTRODUCTION
Gap Problem
Nowadays, many students, graduating from educational
institutions and trying to find a good job, suffer from an
absence of some knowledge they formally must have, but, in
fact, it is missing.1 Such a state of affairs is not new and has
even received the special term “gap problem”.2−5 This notion
refers to a gap between theoretical knowledge (which is usually
a prerogative of universities and other educational institutions)
and practical skills required by various branches of industry.6,7
“To build a bridge” over this gap, worldwide educators are
trying to implement some practicals in the curriculum to
ensure that students are competitive in the job market.8
Nevertheless, this approach cannot completely eliminate the
problem because employers’ requirements are evolving much
faster than the universities’ ability to adopt syllabuses and
primarily renew the equipment used.
The “practice-theory” gap is most sensitive in industries
related to high-tech chemistry, like pharmaceutical, agricul-
tural, and CRO (Contract Research Organization).9 Moreover,
problems concerning the gap are sometimes laid in the first
students’ year. For instance, many higher educational
institutions in Ukraine experience difficulties with insufficient
© 2023 American Chemical Society and
Division of Chemical Education, Inc.
3309
reagents or other equipment to arrange all of the necessary
practices. Thereby, the departments mainly focus on theory
and standard old-fashioned procedures and techniques from
the first years. Besides, the experiments are often undertaken in
groups and not individually, which additionally interferes with
acquiring practical abilities. All of the above-mentioned factors
create a snowball when students lacking basic experimental
skills cannot perform more complex procedures and
consequently develop the ability to solve practical problems;
the modern industry requires this. Although the severity of the
problem varies significantly among the students, chemistry
courses, faculties, universities, and even countries, it is
common and severe enough to keep trying to find a solution.
In this article, our view on its resolution is presented.
Received: December 14, 2022
Revised: June 29, 2023
Published: August 3, 2023
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
Concept/Design of Educational Laboratory Course for
First and Second Year Students
Working for the Ukrainian leading CRO “Enamine Ltd.”, we
have firsthand experience observing the gap problem while
working with undergraduate students. Usually, the students
intern with the company in their 7−8th semesters or after
being awarded a bachelor’s degree. In recent years, we have
noticed that most newcomers do not possess the necessary
qualities that allow them to start solving even simple practical
tasks. Usually, the students knew all the required theoretical
material but could not use it, either because their under-
standing was insufficient or because they lacked general skills
in applying their existing knowledge to practical problems.
Such an obstacle could make the qualified “field” chemists
carry out tedious as well as time- and resource-consuming in-
house training. Obviously, such a state of affairs is not
acceptable. Thereby, a task was set to fix the problem and turn
the theory in students’ minds into practical skills starting right
from their second or even first year in a university. It was
planned to achieve this by creating an innovative cycle of
training works, which is intended to develop primary skills of
independent experimentation, and deepen and fill the gaps in
theoretical knowledge. Hence, in 2017, a joint project of Taras
Shevchenko National University of Kyiv and the “Enamine
Ltd.” was started with the aim to educate the talented and
prone to work in synthetic laboratories students. The start-up
has created a unique tandem of fundamental theoretical
education and practice-oriented training in industrial labo-
ratories, which opened an exceptional opportunity for the
students.
Implementation of such a complex event required careful
design. To reach the goals of the project, it was necessary to
resolve two issues. The first one involved the educational and
methodological part answering what components should be
acquired and how to “embed” them in the course. The second
issue was developing a justified assessment approach, which
should provide a powerful tool for an instructor to reveal
students’ mistakes.
In this way, years of educational and practical experience
allowed us to outline essential techniques/skills to enable
students to start working independently in the laboratory, at
least while carrying out simple experimental procedures.
According to the skills outlined, 12 experimental works for
implementing and reinforcing them were selected. All of the
activities were introduced gradually using the principle of
“easier to more difficult”. Some skills were simultaneously
covered by different lab works of the program, and this indeed
supports better mastering and confident performance of the
activities by the students as they advance through the course.
It should be noted that ideas for some experiments included
in the course were borrowed from the practical tour of the
International Chemistry Olympiads (IChO).10 However, their
design did not allow answering the question “What has a
student done wrong?”, which is undoubtedly essential for the
learning process. Although the works chosen were excellent for
grading students’ performance,11 they did not wholly match
the educational purpose set.
Therefore, the original problems have been transformed to
make a fusion of three components:
(1) educational, including step-by-step mastering the most
demanded essential skills;
3310
pubs.acs.org/jchemeduc Article
(2) analytical, making known all students’ mistakes based on
the results obtained;
(3) competitional, providing an honest and transparent
assessment of student’s performance, which increased
their motivation and allowed one to determine their
level of expertise.
As stated before, grading the students is a fundamental
component of the course design. However, while developing
the course, a problem was faced with how to make the grading
clear, practical, and at the same time simple. In the case of
theoretical knowledge, assessment is a set of well-defined
approaches.12 Meanwhile, accurate and informative assessment
of laboratory skills is still challenging, despite the high
importance of hands-on experience in chemical education.13,14
Partially, the problem has been solved in analytical chemistry
education, where practical skills are graded based on a
deviation of students’ results from master’s values.15 In organic
laboratories, the above-mentioned approach does not work due
to various parameters, which characterize a successful syn-
thesis, e.g., yield, purity, water content, inorganic contami-
nation level, etc.16,17 Furthermore, attempts to improve some
parameters usually lead to downgrading others. For example,
the “yield/product purity” couple could not be graded
separately. Therefore, most organic synthesis practices are
assessed according to the simple “Passed/Not Passed”
principle or using only one synthesis parameter (for example,
yield), which is inadequate. All of the above-mentioned criteria
forced us to search for versatile criteria reflecting how correctly
and successfully every practice task has been fulfilled as well as
methods for determination of this success. As key parameters
for each work, which are easy to check, reasonable time (not
more than 5 h for completely finalizing the practice) and
precise synthetic data (e.g., yield and purity of a product with
their mutual dependence, solvents content) were chosen.
In the following reports, our experience gained from
realizing the course will be shared, in which the current
research is the first in the series.
What Experiment Comes First?
The choice of the topic for the first work in the series was a
responsible and sophisticated task. This experiment should
involve the student in practical chemistry as well as be simple
enough to be conquered by a rookie synthetic chemist.
Considering and trying out various interactions and reaction
setups, our attention was turned to a reaction of 2,4-
dinitrophenylhydrazones formation used as a practical problem
in the 45th IChO.18 The reaction possessed many advantages
over the other candidates. The most vital one is the absence of
the possibility for students to fail when accomplishing the
reaction, which is crucial for the first work, as it must inspire
students with determination to proceed further. Another
valuable feature of the reaction is the simplicity of the synthetic
procedure, which includes simple stirring of reagents in a
beaker at ambient temperature, enabling untrained students to
perform it successfully after a short briefing. It is important that
the labwork allows us to trace students’ mistakes easily and
does not take much time to accomplish.
The preceding explanation clearly indicates the validity of
our laboratory for the pedagogical goals and learning
outcomes. Moreover, this experimental platform provides a
remarkable opportunity to discuss broad aspects of organic
chemistry, such as catalysis, organic compounds identification,
theory of color-structure relationship, drug discovery, and lab
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
safety. This makes it very flexible and applicable to many
teaching needs.
■ PEDAGOGICAL GOALS AND LEARNING
OUTCOMES
Usually, in Ukrainian universities, organic chemistry starts in
the second study year (III semester). Hence, most students
participating in the experiment lacked basic organic laboratory
skills. Moreover, they did not have enough theoretical
knowledge in the field. Of course, students might figure out
the theoretical part of organic chemistry by themselves, which
led to the formation of very diverse groups. Therefore, the
principal goal of the activity was not to give students
fundamental knowledge in organic chemistry but to engage
them in practical work and make them confident while carrying
out standard procedures. At the same time, the necessary
minimum specific theoretical information was provided for
leveling the difference between group members and guarantee-
ing an understanding of “what they are doing”.
Another goal pursued in all our lab works was identifying
possible mistakes that students make and finding out how they
affect the quality of the final product. So, the choice of an
appropriate grading scale is the key milestone for an adequate
assessment of students and for providing them with relevant
feedback about what they have done wrong. Such an approach
significantly increases the motivation of students to continue
their progress in their practical skills. As this work is the first in
the series, students had not been supposed to make
calculations and interim conclusions but simply followed the
experimental procedure. Hence, all mistakes are associated
with the only inaccuracy of students’ performance.
The learning outcomes and pedagogical effectiveness of this
experiment were assessed through students’ experimental
outcomes, such as the yield, purity of products obtained, and
answers to postlaboratory questions. It directly correlated with
their ability to follow prescriptions, performance accuracy, and
understanding of manipulations done. As for specific learning
outcomes, at the end of the experiment, students were
expected to master the following skills:
• weigh out solid compounds
• measure out a specified amount of a liquid substance
• accurately transfer solids and liquids into a container
• use a magnetic stirrer
• perform filtration under reduced pressure
• wash and dry the crude product
• make and record qualitative observations
• use their time productively and efficiently (have time
management skills)
• calculate the yields of a product
■ EXPERIMENT
All glassware and magnetic stir bars are dried in an oven before
use. Students work individually in a fume hood and are
instructed on safe operation before experimenting. The
method for synthesizing hydrazones and the analytical assay
for qualitatively examining the obtained products are given
below.
Synthetic Procedure
Two portions of 2,4-dinitrophenylhydrazine (2,4-DNPH) (200
mg each) are weighed out and transferred to two 50 mL
beakers equipped with a magnetic stir bar and labeled A1 and
3311
pubs.acs.org/jchemeduc Article
A2. Then, using a graduated pipet, 1 mL of phosphoric acid is
added to each beaker, and the mixture is stirred until a
homogeneous mass without large lumps is formed. Next, 1.6
mL of water is added to the beakers, and the mixture is stirred
until a homogeneous suspension is formed. Subsequently,
ethanol is added up to the 10 mL mark in such a way to wash
off all drops and dust from the beakers’ wall.
Preweighed equimolar amounts of aldehydes placed in two
25 mL beakers with labels A1 and A2 are dissolved in about 5
mL of ethanol.
The solutions of the aldehydes are poured into the beakers
with 2,4-DNPH at once (A1 to A1, A2 to A2, respectively)
trying not to moisturize the beakers’ walls. The aldehyde
residues in the beakers were washed off with about 2 mL of
ethanol and transferred to the corresponding beaker. The
mixtures are stirred for 20 min. At this stage, the students
observe how the color of the reaction mixture changes.
During this time gap, the students weigh two glass filters and
two Petri dishes and write A1_XX and A2_XX, where XX is
their number on the students’ list, on Petri dishes.
When the time is up, about 10 mL of water is added to each
beaker, and the mixtures are stirred for an additional 3−5 min.
After that, the stirrers are turned off, and a reaction mixture is
transferred on a filter followed by applying a vacuum to a
Bunsen flask. The residue left in the beaker is washed off with
2−3 portions of water, about 5 mL each.
A vacuum is disconnected; distilled water is poured into the
filter about 4/5 of its height, and the content is mixed with a
spatula until a homogeneous suspension is formed. A vacuum
is applied, and the precipitate is filtered off. Washing is
repeated until the pH of the washing water reaches 7.
Approximately 10 mL of ethanol is poured into the filter.
The content is mixed with the spatula until a homogeneous
suspension is formed. Then, a vacuum is applied, and the solid
is filtered off.
Approximately 10 mL of methyl tert-butyl ether is poured
into the filter. The content is mixed with the spatula until a
homogeneous suspension is formed. A vacuum is applied, and
the solid is filtered off and left on the filter for 5 min. The solid
is fluffed up and left for an additional 5 min.
While the precipitate is getting dry, students wash the 25 mL
beakers.
The filter with the product is weighed and then allowed to
dry for 5 min and weighed again. If the difference between 2
masses is less than 10 mg, drying is considered complete. The
precipitate is transferred to the weighed and marked-up Petri
dish.
The second precipitate is treated in the same manner.
Finally, students weigh the products A1 and A2 in Petri
dishes and write down their masses.
Analytical Assay
Using a glass rod, a minute amount of the products A1 and A2
is placed in the corresponding 25 mL beaker labeled A1 and
A2, and 5 mL of acetone is added to each one. The solutions
obtained have to be yellow with similar intensity.
2 mL of NaHCO3 solution is added to each of the beakers.
Students note their observations.
Next, 1 mL of a NaOH solution is added to each beaker.
Students note their observations.
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
Scheme 1. Principal Synthetic Scheme of the Work
■ HAZARDS
Since low laboratory experience students were engaged in the
work, the elaborated experiment was relatively safe and did not
involve highly reactive, toxic, or highly flammable substances.
It also did not include processes occurring at low and high
temperatures as well as reduced (except filtration) and elevated
pressure. Nevertheless, the overview lecture and prepractice
briefing emphasized that laboratory work requires great care
and attention while performing. Students were also familiarized
with good laboratory practices to work safely in the laboratory
and proper procedures and precautions in handling waste and
chemicals. The importance of informing the supervisor about
potential hazards and accidents that happened was strongly
stressed to students, as well.
The entire experiment was carried out in a fume hood.
Personal protective equipment (disposable nitrile gloves, safety
goggles, and laboratory coat, as well as closed-toe shoes) is
mandatory throughout the laboratory sessions.
Safety profiles of reagents utilized in work are the following.
2,4-Dinitrophenylhydrazine is harmful if swallowed and must
be kept away from heat, hot surfaces, sparks, open flames, and
other ignition sources. Salicylic aldehyde is harmful if
swallowed, may cause skin and severe eye irritation, and is
suspected to cause genetic defects. Concentrated phosphoric
acid is harmful if swallowed and causes severe skin burns and
eye damage. Methyl tert-butyl ether is a highly flammable
liquid and causes skin irritation. Acetone is a highly flammable
liquid, causes severe eye irritation, and may cause drowsiness
or dizziness. 4-Anisaldehyde and solutions of NaOH and
NaHCO3 in specified concentrations are not hazardous
substances for humans.
■ DESIGN AND OVERVIEW OF THE EXPERIMENT
The experiment on hydrazone synthesis typically requires ∼3 h
to complete. The experimental session is preceded by a brief
introductory lecture and a discussion of the planned experi-
ment, highlighting the most significant points of lab processes
and safety concerns. Afterward, the students are sent a
benchmark video of how the experiment is supposed to be
done.
The work is based on a modified procedure of the
preparation of 2,4-dinitrophenylhydrazones by the interaction
of Brady’s-type reagent (solution of 2,4-dinitrophenylhydrazine
in a mixture of phosphoric acid, ethanol, and water) with
salicylic and 4-anisaldehydes (Scheme 1).19,20 Salicylic
aldehyde and 4-anisaldehyde have been chosen as carbonyl
components in the reaction, as they are inexpensive and easily
accessible benzaldehydes giving well-isolable 2,4-dinitrophe-
nylhydrazones in good yields. Moreover, both aldehydes are
liquids at normal conditions, which is very important for the
theoretical part of the work and prevents the determination of
3312
pubs.acs.org/jchemeduc Article
the aldehyde by its physical properties. The formed hydrazones
differ in their color, and this fact may contribute to students’
understanding that even slight changes in a molecule’s
structure may result in a shift in its appearance. In the
experiment, commercially available 2,4-DNPH (98%), salicylic
aldehyde, and 4-anisaldehyde (98+%) were used without
additional purification. Hazardous methanol and sulfuric acid,
common reagents for such interactions, are not used in work.
While the experiment was elaborated, the intention was to
get informative and repeatable results rather than finding the
best reaction conditions that provide excellent yields. There-
fore, in contrast to the classic method, our methodology
utilized equimolar amounts of the reagents. This enables us to
assess the weighing skill of a student (both under- and
overweighing) (vide inf ra).
One should note that students are given exact masses of the
aldehydes preweighed by qualified personnel, prepare their
solutions, and transfer them to the reaction mixture. Besides,
the students weigh out 2,4-DNPH and perform the syntheses
of hydrazones 4 and 5 in accordance with the procedure (see
Supporting Information). Such an approach eliminates double-
weighing errors, leading to results that are challenging to figure
out.
A simple analytical experiment was added to the practicum
to demonstrate general principles of identifying organic
compounds via a test reaction on a functional group. As one
can see, the hydrazones feature the acidic NH group, in which
deprotonation must lead to a notable color change of the
solution. Even though hydrazones 4 and 5 have very close pKa
values of 10.84 and 11.03, respectively,21,22 the proper
concentration of NaHCO3 solution to selectively deprotonate
product 4 and thus cause a color shift was found. At the same
time, the subsequent addition of a solution of NaOH as a
much stronger base results in deepening the color of both
solutions (Figure S1). Importantly, color changes must be
noticeable, and solutions remain optically transparent through-
out the test. Exactly in search of the optimal pair to achieve this
effect, many benzaldehydes were sorted out.
When the work is accomplished, the students are supposed
to hand in their products to be put on Petri dishes. Taking the
masses of the obtained products, the students also calculate
their yields. Then, the students fill out the “Paper for answers”
(mass and yield of the products, their observations regarding
analytical assay), which is assessed by an experiment
coordinator. The experiment typically requires approximately
3 h to complete.
The next day, students receive oral feedback on their work
including an analysis of mistakes they have made. Subsequent
discussion, if needed, is also held.
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
Figure 1. Visualization of the equation calculating the grade for the yield/purity couple.
■ GRADING APPROACH
Students’ works are graded quantitatively according to a
worked-out scale, including a complex mark for the yield and
purity of the products (0−10 pts for each hydrazone); dryness
of the products handed in (weight loss on drying, 0−1 pts for
each hydrazone); students’ performance in doing the test
reaction (0−1 pts).
Understanding that yield and purity of the products are two
of the most influential parameters, an idea, which was
previously used in the International Chemistry Olympiads,
was adapted to our practice.23 It enabled the calculation of
marks and reflected all possible combinations of these
parameters. An equation has been written for all calculations
used throughout the article (see the Supporting Information).
For instance, the highest grade a student obtains corresponds
to a yield in the 75−99% range and purity >98%. While either
of the parameters is worsened, the mark is also gradually
falling; if purity gets 50% and lower and the yield is out of the
30−120% range, a student does not receive any points for this
component of the total mark. When one takes these ranges and
values into account, the equation is “visualized” to ensure
satisfactory results (Figure 1A). It is worth mentioning that the
equation is flexible and adjustable to particular needs by simply
altering the variables.
One can see that the obtained pyramidlike shape has a small
area on the top relating to a 10 pt grade when the highest
requirements are met. Three faces and two irregular surfaces of
the shape have specific inclinations, which depend on the
starting variables’ values and indicate the grade fall rate. The
back face is perpendicular to the horizontal surface due to the
maximum possible purity of 100%. Isolines in Figure 1B show
the dependence of yield/purity values on the location of the
grade on the surface.
For the part “dryness”, students receive 1 point if a solid they
handed in loses less than 0.2 g of the mass upon drying.
Otherwise, the value is 0 pts.
Students’ performance in carrying out test reactions was
evaluated based on their reports: 1 point was granted in the
case of correct answers. Otherwise, students obtain 0 pts.
Penalties (e.g., time ones) were not applied when grading
the students. Nevertheless, they can be used during the
3313
pubs.acs.org/jchemeduc Article
adaptation of the experiment to competitive activities, like
Olympiads, tournaments, etc.
Detailed grading guidance with complete experimental data
and all equations for calculating the total mark are given in the
Supporting Information.
■ TYPICAL STUDENTS’ MISTAKES
While designing the experiment, it was concluded that efficient
analysis of results submitted by the students is only possible if
one understands the places the students stumble over and how
these missteps affect the experiment’s outcome. In this regard,
an attempt was made to envisage possible mistakes or
inaccuracies in students’ performance. Further, it was
investigated how such procedure deviations impact the time
required to accomplish the experiment, the yield, and purity of
the products. A list of the foreseen mistakes is as follows:
1. 2,4-DNPH was weighed wrongly or partially transferred
to a beaker
2. Incomplete dissolution of the starting aldehydes
3. Partial transfer of the aldehydes (the solution was spilled
out or the step of washing off the residues was missed)
4. The acid was not added to the reaction mixture
5. The step of adding water after completion of the
reaction was missed
6. The filtration procedure was broken (e.g., the residue
was washed insufficiently)
7. Inadequate drying of the products
8. High concentration of the products while performing the
analytical assay
One would like to note in advance that all predicted
mistakes have been encountered to a greater or lesser extent
while observing students’ performance.
Tryout of the Experimental Procedure
First, the experimental procedure was subjected to “examina-
tion” by qualified synthetic chemists with the purpose of
finding out the yields and purities of the products it gives,
provided that everything was done correctly. A total of 14
qualified chemists were engaged in the test, each making 7
attempts (98 in total). Both yield-without-isolation (amount of
the products on the filter) and preparative yield (amount of the
products removed from the filter) were measured (Table 1).
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
Table 1. Benchmark Yields
Product
Parameter 4 5
Yield-without-isolation (mean value), % 85 87
Range, % 82−89 83−90
Preparative yield (mean value), % 80 78
Range, % 77−85 70−85
Purity (according to 1H NMR) ≥98 ≥98
The yield-without-isolation of 4 is slightly lower, probably due
to its higher solubility. The coarse-grained crystalline
precipitate dries quickly and is easily transferred to a Petri
dish with less than 5% losses. Similar losses during the transfer
of 5 are almost twice as high, which leads to a lower
preparative yield, despite higher yields without isolation. The
reason is that compound 5 is a much finer solid. It clogs up the
pores of the filter, gets electrified, and sticks to the walls. This
makes it more challenging to work with the hydrazone, thus
increasing losses.
1
H NMR spectra of the products obtained in these
experiments were also recorded (Figure S2). One can conclude
that the procedure is valid and gives pure hydrazones in high
yields while following the synthetic prescriptions. The yields
and spectra were taken as standard ones and used in further
analyses.
Less 2,4-DNPH than Needed
Inspection of the yields-without-isolation obtained by gradually
reducing the amount of 2,4-DNPH in the reactions (in % of
stoichiometric) indicates that in all cases 100% conversion of
2,4-DNPH into the product is achieved. At the same time, an
excess of the aldehydes is easily removed during filtration and
washing the precipitate. The product yield decreases almost
linearly with a decrease of the 2,4-DNPH amount utilized in
the reaction. Simultaneously, the purity of the products
remains >98% (1H NMR) and is independent of the yield
(Figure 2). Thus, the underweighting error causes a reduction
in product yield but does not affect its purity.
Figure 2. Dependence of the yields (lines) and purities (columns) of
the products 4 (orange) and 5 (red) on an amount of 2,4-DNPH
used in the interaction.
3314
pubs.acs.org/jchemeduc Article
More 2,4-DNPH than Needed
Unlike the previous case, the complete conversion of the
aldehydes was spotted while exploring the yield-without-
isolation in reactions with more than an equimolar amount of
2,4-DNPH. Meanwhile, an excess of 2,4-DNPH worsens the
quality of the product, as the former is a constant admixture to
the final hydrazones. The last fact may lead to calculated
“imaginary” yields exceeding 100% even after drying. The
reddish color of the contaminated product is more reliable
evidence of the overweighing mistake made by students. It is
clear that assessing such a parameter as “yield” does not make
sense after passing a certain threshold. Thus, when checking
students’ results, it was reasonable to assess only purity if the
yield value goes past a limit of 120%. Corrected yields
(regarding purity) and the content of hydrazones in the
mixture are given in Figure 3.
Figure 3. Dependence of corrected yields (lines) and purity
(columns) of hydrazones 4 (orange) and 5 (red) on an excess of
2,4-DNPH used in the reaction.
Since 2,4-DNPH is partially soluble in both water and
ethanol, its content in the final mixture does not increase
linearly. Moreover, contaminated products feature a higher
solubility, and the washing step additionally reduces the yield.
Elevated viscosity is another important peculiarity of the
impure products, complicating filtration. Thus, the experiments
require more time to complete. 1H NMR spectra of the
products also clearly indicate contamination, as they contain
notable signals of 2,4-DNPH (Figure S3).
Partial Transfer of the Aldehydes (Incomplete Dissolution
of the Aldehydes)
These mistakes are tightly connected with students’ inaccuracy
and may significantly affect both the yield and purity,
decreasing the quality of the products. The procedure
violations closely relate to the preceding issue and have similar
consequences.
Acid Has Not Been Added to the Reaction Mixture
An acid as the catalyst in the reaction facilitates the process and
speeds it up. Thereby, if the acid was missed, then one should
expect the partial conversion of the starting products. It was
determined that only 20% of the starting material reacted
during the experiment (20 min), and the isolated product was
mainly 2,4-DNPH (Figure S4). The reaction mixture features
red color, and filtration and washing take longer due to
elevated viscosity.
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
The Filtration Procedure Was Broken
Insufficiently washed precipitate contains acid traces. This
results in slower drying and notable widening of the NH signal
in the 1H NMR spectrum (Figure S5). In addition, a lack of
precipitate mixing during washing and insufficient volume of
the solvent used for washing also slow down the drying of the
precipitate. As a result, there is a significant decrease in the
mass of the product upon drying; signals of solvent protons
can be present in the 1H NMR spectra as well.
Inadequate Drying of the Products
According to the synthetic procedure, a partially dry solid
product must be fluffed up and left on the filter for an
additional 5 min. Otherwise, due to high resistance to the
airflow, drying proceeds slower and takes more than 10 min.
Furthermore, missing this step results in a dense “tablet” firmly
adhering to the walls and bottom of the filter, which
complicates the transfer of the product to a Petri dish, leading
to a decrease in yield. This mistake can also be a result of the
missteps discussed before.
The Step of Adding Water after Completion of the
Reaction Has Been Missed
This mistake does not significantly affect the yield and purity of
the products. However, the resulting precipitate is a delicate
substance that substantially increases the duration of the
filtration step.
High Concentration of the Products while Performing the
Analytical Assay
It complicates conducting the analytical test due to similar
colors and shades in both beakers. Nonetheless, preparing new
solutions with suitable concentrations can easily overcome this
mistake.
Generalized Data on the Impact of Errors on
Yield/Purity/Time
Table 2 includes information on how the discussed mistakes
affect the products’ quality and the experiment’s duration. One
Table 2. Effects of the Mistakes on the Quality of the
Product and the Time of the Experimenta
Mistake Yield Purity Time
More 2,4-DNPH than needed + +
Less 2,4-DNPH than needed + −
Partial transfer of the aldehydes + ±
Acid was not added + +
Precipitate was washed improperly ± −
Water was not added − −
a
Notes: “+”: mistake affects significantly; “−”: mistake has no effect
on a parameter; “±”: mistake has an impact in some cases.
can see that most mistakes result in yield deviation from “true”
values. Also, overweighing 2,4-DNPH and missing a step of
acid addition are the worst missteps, as they contribute to all
three parameters. Indeed, the collected data provide a powerful
tool for analyzing students’ works and may support the
students to succeed in further experimental sessions.
■ RESULTS AND DISCUSSION
Over a three year duration (2018 to 2020), 10 cohorts of
undergraduate students have successfully performed the
laboratory experiment (typically 20 students in a group, 196
students in total). Students in a cohort were divided into two
pubs.acs.org/jchemeduc Article
groups undertaking the experiment in two identically equipped
laboratories. Students experimented in an individual fume
hood. Before they started working, information about the
laboratory layout and the location of devices they needed was
shown to the students.
The students’ performance has been assessed based on the
quality of their resultant products, submitted reports, and
observations made by the instructor. All collected information
was categorized and statistically processed.
Dryness, Purity, and Yield of the Products
The purity and the yield of the reaction products are the most
important characteristics of a successful synthesis. The
presence of residual solvents affects the product quality to a
lesser extent as it can be improved by simple drying. Analysis
of these parameters allows one to deduce how students can
follow the experimental procedure and their accuracy in the
prescribed activities.
To examine the dryness of products submitted by students,
the solids were additionally dried overnight in an oven at 40
°C. The difference in masses was considered the moisture
content. It was established that more than half participants
provided relatively dry hydrazone 4 (Figure 4). In the other
Figure 4. Moisture content in hydrazones 4 (orange) and 5 (red)
+ provided by students.
−
−
25% cases, the solvent content did not exceed 20%. The
+
situation with hydrazone 5 was found to be the opposite. Less
+
than 10% of the students hand in dry product 5, while ∼67%
+
provided the product containing more than 50% residual
moisture. As shown, the finer hydrazone 5 is much more
difficult to wash. Thus, it dries slowly on the filter. It is
challenging for most students to assess its dryness properly and
handle the precipitate successfully, most likely due to deviation
from the procedure.
The products’ purity was examined using the 1H NMR
method by the ratio of integral intensities of hydrazones’
aromatic proton signals and NH2 protons of 2,4-DNPH. The
application of HPLC was unsuitable in this case because this
method gives inaccurate results due to the difference in
attenuation coefficients of the investigated products. The
results evidence that 41% and 36% of the students obtain
hydrazones 4 and 5, respectively, with excellent purity of more
than 98% (Figure 5). The rest of the samples comprise a
different amount of impurities, which are 2,4-DNPH and
3315 https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
Figure 5. Purity of hydrazones 4 (orange) and 5 (red) provided by
students.
residual solvents. According to the series of experiments
discussed earlier, possible causes are weighing errors, partial
transfer of the aldehydes, and improper washing/drying of the
product.
Adjusted yields concerning purity are shown in Figure 6. A
tiny percentage of students obtain yields approaching or
Figure 6. Yields of hydrazones 4 (orange) and 5 (red) provided by
students.
exceeding the benchmark values. The distribution of yields in
the case of 4 differs from 5 and is shifted toward higher yields
due to the difficulty of extracting hydrazone 5 from a glass
filter.
However, as mentioned above, the yield and purity are
interrelated parameters and should preferably be assessed
together. Such an approach amplifies the analysis of mistakes
and gives more precise feedback to the students. Figure 7
represents a plot of “purity” vs “yield” with marked areas
connected with specific mistakes.
Difference in Yields of Hydrazones 4 and 5
Comparing the yields that the same student obtained for
hydrazones 4 and 5 would allow us to assess students’
professionalism and accuracy in performing the experiment. As
one can see from Figure 8, more than 50% of students fall in
the different ranges of 0−5%, which is typical for careful work
3316
pubs.acs.org/jchemeduc Article
Figure 7. Analysis of “purity” vs “yield” plot.
Figure 8. Difference in yields of hydrazones 4 and 5 as an indicator of
students’ carefulness.
performance as the yields of the hydrazones are similar. At the
same time, a difference of more than 20% is observed for about
10% of participants. This is a clear marker of blunders made by
students during the synthesis/isolation of the product.
Performance of the Analytical Assay
An analytical examination of hydrazones was the most exciting
part for the students. They eagerly carried out the test and
were curious about the difference in the hydrazones’ behaviors.
Statistically, a large majority of the students fulfilled the assay
successfully by following the procedure (Figure 9). However,
explaining the test results was a tough nut to crack for the
students, apparently because of insufficient theoretical back-
ground.
Time Management
Time management is a crucial issue for modern practical
chemistry. Nowadays, synthetic chemists experience ever-
growing pressure to be exponentially more productive. Any
time wasted or unused efficiently will inevitably increase the
turnaround time and thus income loss. On the other hand, the
student’s lab work must be strictly regulated in time, which
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education pubs.acs.org/jchemeduc Article

Table 3. Timeline of the Synthetic Procedure

Average Time before Average Time Needed for an
Finishinga, h:min Activitya, min ± min
Experienced Experienced
Activity Chemist Student Chemist Student
Weighing out 2,4- 0:04 0:10 4±1 10 ± 5
DNPH
Addition of the 0:06 0:14 2±1 4±2
acid and water
Addition of ethanol 0:10 0:19 4±1 5±2
Addition of the 0:12 0:21 2±1 2±1
aldehyde
solutions
Stirring the 0:32 0:41 20 20
reaction mixtures
Addition of water 0:33 0:42 1±1 1±1
Filtration of a 0:38 0:48 5±2 6±4
precipitate 1
Figure 9. Performance of students in doing the analytical test (blue, Washing with water 0:43 1:08 5±2 20 ± 6
succeeded in the assay; magenta, failed the assay). to reach pH 7
Washing with 0:45 1:13 2±2 5±3
ethanol
makes it possible to use in a rigorous organized educational Washing with 0:48 1:18 3±1 5±2
process. MTBE
In this regard, much emphasis was put on doing optional Drying on the filter 0:58 1:32 10 ± 2 14 ± 5
activities in our lab work between other synthetic procedures. Filtration of a 1:03 1:38 5±2 6±3
precipitate 2
In particular, while a residue is drying, students wash beakers
Washing with water 1:09 2:02 6±2 24 ± 7
or prepare solutions of the obtained compounds and perform to reach pH 7
the analytical assay. Washing with 1:11 2:06 2±1 4±2
Besides, the time students need to accomplish each step of ethanol
the procedure was compared to that needed by experienced Washing with 1:14 2:12 3±1 6±3
chemists (Table 3). The aim was to find out what operations MTBE
Drying on the filter 1:24 2:29 10 ± 2 17 ± 5
cause more difficulty for rookies. As one can see, washing the
Transfer of the 1:34 2:40 10 ± 2 11 ± 3
precipitates appeared to be the most strenuous activity for the products to Petri
students, and it takes four times as long as the washing takes dishes and
for experienced chemists. Weighing out 2,4-DNPH, drying the weighing them
precipitates, and performing the test reaction also cause some Carrying out the 1:46 3:00 12 ± 2 20 ± 5
test reaction
difficulties. However, all the mentioned activities are practice Noting results of 1:51 3:11 5±1 11 ± 6
dependent, and their improvement in further experiments is the work
possible. Total time 1:51 ± 0:11b 3:11 ± 0:38b
In regard to total time, most students required ∼2.8−3.2 h a
Notes: The results were received in our laboratories and their use in
to accomplish the experiment, which is 1 h more than the time other circumstances may be unsuitable. bThe real deviations are given.
needed by a qualified chemist (Figure 10). A small number of
students were out of the range.
Assessment of the Students
The students’ performance and the work’s effectiveness were
assessed quantitatively as described in Design and Overview of
the Experiment. An overview of students’ grades (in % of total)
is given in Figure 11: blue columns indicate synthetic part
(yield and purity); pink columns represent total grade. The
diagram reveals a slightly different distribution of marks within
the groups mentioned above, with total grades shifted toward
lower values. This is mainly attributed to inadequate drying of
the products and, consequently, points to loss. Nonetheless,
the vast majority of students received acceptable (>60%) to
excellent grades and thus have mastered skills coming under
the spotlight in this work. This fact is ground to promote them
further through the course. Moreover, the students’ good
results evidently demonstrate the applicability of the developed
practical platform to training newcomers in laboratory
techniques. Figure 10. Distributions of the students depending on the time they

■ SUMMARY AND CONCLUSIONS

Operationally simple lab work has been devised for under-
need to accomplish the experiment.

graduate first year students without any or little laboratory

3317 https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education
Figure 11. Distribution of the students’ grades for the synthetic part
(blue) and total grade (pink).
experience. The work is based on the well-known interaction of
2,4-dinitrohydrazine with benzaldehydes, subsequent isolation
of the hydrazones formed, and their characterization by the
test reaction. The experiment does not employ highly toxic and
flammable substances and lasts approximately 3 h. It requires
minimal setup and cheap commercially available reagents and
solvents; all operations are performed at room temperature.
The listed advantages make the work an excellent training in
synthetic chemistry and product analysis for newcomers.
Moreover, aimed at reinforcing the development of students’
primary laboratory skills, the work successfully deals with the
goal as more than 70% of students earn 80−100% of the total
grade. The observations of the supervisor, as well as verbal
feedback, gathered from the students provide evidence that the
participants have a deeper understanding of laboratory
processes and became more confident while operating the
laboratory equipment.
In addition, the experiment is flexible and adjustable to meet
the requirements of other tasks. For instance, it can be easily
extended by introducing the multi-outcome experiment
approach allowing students to identify which aldehyde they
are working with by analysis of spectral data (1H NMR, IR,
etc.). Finally, although this experimental work was designed for
teaching purposes, it can be adapted for various competitions
(e.g., olympiads). Anyway, all the applications will be
supported by a carefully designed comprehensive grading
system.
■
*
ASSOCIATED CONTENT
sı Supporting Information
The Supporting Information is available at https://pubs.ac-
s.org/doi/10.1021/acs.jchemed.2c01214.
Spectral data; algorithm of grading system (PDF)
Guide to the laboratory work (PDF)
Student lab manual and lab report (PDF)
Video showing experiment overview (source of music in
video: https://www.youtube.com/watch?v=
ROPJPhACN7w; CC BY 3.0 license, https://
creativecommons.org/licenses/by/3.0/) (MP4)
3318
pubs.acs.org/jchemeduc Article
■ AUTHOR INFORMATION
Corresponding Authors
Sergey V. Ryabukhin − Enamine Ltd., 02094 Kyiv, Ukraine;
Taras Shevchenko National University of Kyiv, 01033 Kyiv,
Ukraine; Institute of Organic Chemistry, National Academy
of Sciences of Ukraine, 02094 Kyiv, Ukraine; orcid.org/
0000-0003-4281-8268; Email: s.v.ryabukhin@gmail.com
Dmitriy M. Volochnyuk − Enamine Ltd., 02094 Kyiv,
Ukraine; Taras Shevchenko National University of Kyiv,
01033 Kyiv, Ukraine; Institute of Organic Chemistry,
National Academy of Sciences of Ukraine, 02094 Kyiv,
Ukraine; orcid.org/0000-0001-6519-1467;
Email: d.volochnyuk@gmail.com
Authors
Valeriya G. Makhankova − Enamine Ltd., 02094 Kyiv,
Ukraine; Taras Shevchenko National University of Kyiv,
01033 Kyiv, Ukraine; orcid.org/0000-0002-0012-5108
Alexander Yu. Lyapunov − Enamine Ltd., 02094 Kyiv,
Ukraine; Taras Shevchenko National University of Kyiv,
01033 Kyiv, Ukraine
Liliia S. Muravska − Enamine Ltd., 02094 Kyiv, Ukraine
Olexander M. Vitsenko − Enamine Ltd., 02094 Kyiv,
Ukraine
Complete contact information is available at:
https://pubs.acs.org/10.1021/acs.jchemed.2c01214
Notes
The authors declare no competing financial interest.
■ ACKNOWLEDGMENTS
The work was supported by Enamine Ltd. and personally by
Andrey A. Tolmachev. The authors also thank UkrOrgSyntez
Ltd. and its director Serhii I. Dovgopolyi personally for help in
organizing the practice. Also, the authors acknowledge Dmitry
A. Lega for his help with manuscript preparation.
■ REFERENCES
(1) Wang, L. Closing The Skills Gap. Chem. Eng. News 2012, 90
(24), 49−51.
(2) Furman Shaharabani, Y.; Yarden, A. Toward narrowing the
theory-practice gap: characterizing evidence from in-service biology
teachers’ questions asked during an academic course. International
Journal of STEM Education 2019, 6 (1), 21.
(3) Greenway, K.; Butt, G.; Walthall, H. What is a theory-practice
gap? An exploration of the concept. Nurse Education in Practice 2019,
34, 1−6.
(4) McGarr, O.; O’Grady, E.; Guilfoyle, L. Exploring the theory-
practice gap in initial teacher education: moving beyond questions of
relevance to issues of power and authority. Journal of Education for
Teaching 2017, 43 (1), 48−60.
(5) Foltean, F. S. Bridging marketing theory - practice gap to
enhance firm performance: Introduction to the special issue. Journal of
Business Research 2019, 104, 520−528.
(6) Carr, W. The Gap between Theory and Practice. Journal of
Further and Higher Education 1980, 4 (1), 60−69.
(7) Bowman, R. E. A secondary course in industrial chemistry. J.
Chem. Educ. 1927, 4 (3), 346.
(8) Casanova, J.; Tunstad, L. A Practical Test of Organic Chemistry
Laboratory Skills. Chemical Educator 1996, 1 (1), 1−4.
(9) Kondo, A. E.; Fair, J. D. Insight into the Chemistry Skills Gap:
The Duality between Expected and Desired Skills. J. Chem. Educ.
2017, 94 (3), 304−310.
https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319
Journal of Chemical Education pubs.acs.org/jchemeduc Article

(10) International Information Centre of the International Chemistry

Olympiads Home Page; https://www.icho.sk/ (accessed 2022-08-12).
(11) Kmentova, I.; Almassy, A.; Feriancova, L.; Putala, M. Assessing
Basic Separatory Technique Skills in an Organic Experiment Based on
an Incomplete Haloform Reaction. J. Chem. Educ. 2020, 97 (4),
1139−1144.
(12) Goubeaud, K. How is Science Learning Assessed at the
Postsecondary Level? Assessment and Grading Practices in College
Biology, Chemistry and Physics. Journal of Science Education and
Technology 2010, 19 (3), 237−245.
(13) Hofstein, A. The laboratory in chemistry education: thirty years
of experience with developments, implementation, and research.
Chemistry Education Research and Practice 2004, 5 (3), 247−264.
(14) Chen, H.-J.; She, J.-L.; Chou, C.-C.; Tsai, Y.-M.; Chiu, M.-H.
Development and Application of a Scoring Rubric for Evaluating
Students’ Experimental Skills in Organic Chemistry: An Instructional
Guide for Teaching Assistants. J. Chem. Educ. 2013, 90 (10), 1296−
1302.
(15) Stone, D. C. Should students be graded on accuracy and
precision? Assessment practices in analytical chemical education.
Anal. Bioanal. Chem. 2017, 409 (7), 1719−1724.
(16) Pirrung, M. C. Handbook of Synthetic Organic Chemistry, 2nd
ed.; Academic Press, 2017.
(17) Isac-Garcia, J.; Calvo-Flores, F. G.; Dobado, J. A.; Martinez-
Garcia, H. Experimental Organic Chemistry. Laboratory Manual;
Academic Press, 2015; DOI: 10.1016/C2015-0-00644-X.
(18) Sirota, A.; Ed. The competition problems from the International
Chemistry Olympiads; Vol. 3, 41st−45th IChO (2009−2013);
IUVENTA, 2014.
(19) Brady, O. L.; Elsmie, G. The Use of 2:4-Dinitrophenylhy-
drazine as a Reagent for Aldehydes and Ketones. Analyst 1926, 51
(599), 77−78.
(20) Brady’s test for aldehydes and ketones; https://edu.rsc.org/
experiments/bradys-test-for-aldehydes-and-ketones/549.article (ac-
cessed 2022-07-11).
(21) Jones, L. A.; Mueller, N. L. Spectrophotometric Studies of
Some 2,4- Dinitrophenylhydrazones. IV. The pKa’s of m- and p- Recommended by ACS
Substituted Aromatic Carbonyl Derivatives. J. Org. Chem. 1962, 27
(7), 2356−2360.
(22) Alizadeh, K.; Amraie, A. Electronic absorption spectroscopic From Soybeans to Tofu: The Underlying Chemistry
behavior and acidity constants of some new dinitrophenylhydrazone Bingxing Wang, Donglin Zhao, et al.
derivatives. Spectrochim. Acta, Part A 2015, 147, 67−72. AUGUST 17, 2023
(23) 47th International Chemistry Olympiad, Baku, Azerbaijan, July JOURNAL OF CHEMICAL EDUCATION READ
20−29, 2015. Practical Examination, July 23; http://icho2015.msu.az/
uploads/files/IChO-2015_Practical_Exam_Official_Grading_ A Hidden Aspect of Laboratory Safety
Scheme-final.pdf (accessed 2022-08-23).
Michael W. Burand.
AUGUST 25, 2023
JOURNAL OF CHEMICAL EDUCATION READ

An Experimental Approach with a Twist: Helping High

School Students to Understand the Concept of Limiting
Reactant
Valeria Edelsztein.
MARCH 08, 2023
JOURNAL OF CHEMICAL EDUCATION READ

Lipids and Proteins: Worksheets, Concept Maps, and Safe

Experiments
Kateřina Trčková, Roman Maršálek, et al.
JUNE 07, 2023
JOURNAL OF CHEMICAL EDUCATION READ

Get More Suggestions >

3319 https://doi.org/10.1021/acs.jchemed.2c01214
J. Chem. Educ. 2023, 100, 3309−3319