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Jun 28, 2017 - ABSTRACT: Analytical method transfer (AMT) and dissolution testing are important topics required in industry that should be taught in a...
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Teaching Analytical Method Transfer through Developing and Validating Then Transferring Dissolution Testing Methods for Pharmaceuticals Irene Kimaru,*,† Marina Koether,‡ Kimberly Chichester,† and Lafayette Eaton† †

Department of Chemistry, St. John Fisher College, 3690 East Avenue, Rochester, New York 14618, United States Department of Chemistry and Biochemistry, Kennesaw State University, 370 Paulding Avenue Northwest, MD#1203, Kennesaw, Georgia 30144, United States

‡

S Supporting Information *

ABSTRACT: Analytical method transfer (AMT) and dissolution testing are important topics required in industry that should be taught in analytical chemistry courses. Undergraduate students in senior level analytical chemistry laboratory courses at Kennesaw State University (KSU) and St. John Fisher College (SJFC) participated in development, validation, and transfer of two dissolution testing methods for pharmaceuticals. These experiments were transferred between the two schools and underwent method equivalency testing by a different set of undergraduate students to validate the AMT. Student learning outcomes addressed at both SJFC and KSU include (1) significant learning gains across all 3 years of the study; (2) confidence in method development, writing standard operating procedures, and laboratory transfer of validated methods; (3) increased understanding of AMT and dissolution testing; (4) ability to operate dissolution testers, UV−vis, and flame atomic absorption spectrophotometers; and (5) improved knowledge and understanding of figures of merit and statistical analysis. KEYWORDS: Analytical Chemistry, Upper-Division Undergraduate, Curriculum, Instrumental Methods, Hands-On Learning/Manipulatives, Applications of Chemistry, Communication/Writing, Drugs/Pharmaceuticals, Spectroscopy

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receiving laboratory is still able to get the same results, within experimental error, as the originating laboratory and demonstrate equivalence between the two laboratories mean responses. This statistical analysis of whether the method is equivalent to the original one after the method transfer is currently not taught at the undergraduate level. AMT is a subtopic of method equivalency assessments as described recently in the Perspectives Section of the December 2009 ACS Journal of Analytical Chemistry,6,7 and earlier as a featured article,8 clearly indicating that this knowledge is necessary for the Analytical Chemist. Typically, college students graduate with a nonexistent foundation in this type of assessment and are unaware of the thorough analysis required for an AMT. This article brings this recent advancement in STEM disciplinary knowledge9−16 into the undergraduate experience. AMT requires the application of skills involving method development, method validation, and method equivalency. All students obtaining a degree in chemistry take quantitative analytical chemistry and an upper level analytical chemistry course such as instrumental analytical chemistry. In this sequence of courses, the learning outcomes of method development, method validation (i.e., figures of merit: accuracy, precision, specificity, selectivity, sensitivity, repeatability, reproducibility, linearity, range, detection limit, quantitation limit, robustness, and ruggedness), quality control, quality assurance, and the associated statistics should be taught;

nalytical method transfer (AMT) is conducted globally in today’s analytical and pharmaceutical industries to ensure product reproducibility and quality control. This topic has been addressed by key regulatory bodies such as the American Association of Pharmaceutical Scientists (AAPS), the Food and Drug Administration (FDA), and European Union (EU) regulatory authorities.1 AMT is rarely taught in undergraduate analytical chemistry courses because there is currently no curriculum available that specifically addresses this topic. Consequently, BS chemistry graduates who obtain jobs in the industry typically enter the workplace lacking the necessary background and must be trained on the job. This is unfortunate given that this knowledge is a current global need in the industry. This paper addresses this deficit as it is a major topic at conferences and in the trade journals, for example, American Chemical Society Short Course in Philadelphia in August 2016,2 PITTCON Symposia in 2007,3 LCGC,4 and Pharmaceutical Technology.5 In general, AMT is the transfer of any analytical method from the originating laboratory to another laboratory. In industry, method transfer studies are used to ensure the successful transfer of an analytical method from the development “originating” laboratory to the manufacturing quality control “destination” laboratory responsible for releasing the product. It is the analytical chemist who develops the method and conducts the method validation. Once transferred to the new lab, another analytical chemist repeats the method validation in their laboratory and thus follows the written procedure to try replicate the figures of merit. The main goal of the method transfer study and statistical analysis is to demonstrate that the © XXXX American Chemical Society and Division of Chemical Education, Inc.

Received: November 12, 2016 Revised: June 1, 2017

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Consequently, the pharmaceutical industry has to provide comprehensive employee training for new hires.28 For these reasons, there is need to introduce dissolution testing in the undergraduate chemistry curriculum. Six two-day courses on the subject of dissolution testing are taught at Agilent workshops yearly29 and courses on dissolution testing are also available through the United States Pharmacopeia (USP). So far, the number of chemical educational literature available to guide the instructor is limited.30,31

however, there is a lack of chemical educational journal articles to guide the instructor.17−25 This study presents the curriculum concepts of AMT and dissolution testing, two practices that are routinely performed in the pharmaceutical industries by B.S. level Chemists. These two concepts were introduced into analytical chemistry courses at St. John Fisher College (SJFC) and Kennesaw State University (KSU). Undergraduate students at these two distant schools were engaged in developing dissolution testing methods for pharmaceuticals products, writing standard operating procedures (SOPs), performing method validation, transferring validated analytical methods, and assessing the validation parameters for comparability by using equivalency testing. This setting provided a real world experience for the undergraduate students who do not often have the opportunity to perform work that is done in industry. While the current work could merely focus on the transfer of an UV−vis method, the experiments involving dissolution testing will offer students the opportunity to gain experience using a very common technique used by B.S.-prepared chemists in the pharmaceutical industry. Dissolution testing is a key quality control procedure that determines if a pharmaceutical drug can release its active ingredient and become available in solution for absorption in the gastrointestinal track. The dissolution tester is used to simulate the dissolution in the stomach and small intestines to indicate the bioavailability of the drug. Since most drugs are absorbed in the upper small intestines, it is important to verify that the drugs are released from their dosage forms prior to reaching the absorption window. Thus, a test is necessary to know when the majority has dissolved. The apparatus consists of six inert vessels and corresponding rotating paddles or baskets. A suitable dissolution medium is selected to mimic the stomach or intestine environment. During early drug product development, in vitro dissolution testing is used to evaluate formulation prototypes of the active pharmaceutical ingredient (API). The results are often compared to in vivo bioavailability data to draw correlations. Once the dissolution methods are established, they are used to study different formulations, different batches, fresh products versus stored products, old processes versus new processes, and a generic product versus the innovator product. This is accomplished through comparing dissolution profiles and determining the difference and similarity factors, denoted by f1 and f 2 and defined by eqs 1 and 2:26 n

GENERAL DESCRIPTION OF THE STUDY Two new topics were introduced in upper level undergraduate analytical chemistry courses to address two current topics that are prevalent in the pharmaceutical industry, those of AMT and dissolution testing. The main objective of adding these topics is to provide undergraduate students the opportunity to learn how dissolution testing and AMT are used in the pharmaceutical industry while learning important fundamental principles taught in analytical chemistry. The method development, method validation of two laboratory experiments were completed by undergraduate students in analytical chemistry laboratory courses at two different schools: KSU and SJFC. These methods were transferred between the two schools and underwent method equivalency testing by a different set of undergraduate students to validate the AMT by statistical comparison of the figures of merit. Only statistical analysis would determine if the AMT was successful. While it may seem unnecessary to have two schools, each instructor does have their own educational bias and to create a nonschool specific AMT that would be transferable to any school, the use of two dissimilar schools allows for a broader appeal of the final product than what would be produced from one school alone. The experiments involved the dissolution of pharmaceutical dosage forms with the development of dissolution curves (release rates) as described by the USP and the FDA. The concentration of the active pharmaceutical ingredient in the dissolution media at each time point after immediate filtration was determined by student validated UV−vis and flame atomic absorption spectroscopic methods.

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STUDENT FOCUSED LEARNING STRATEGY Each college had students in their respective analytical laboratory course: (1) develop a transferable analytical method, (2) receive and validate a transferred analytical method, and (3) receive a validated analytical method that originated at their school. Thus, the students at one college were taught the basics of dissolution testing and each given a USP monograph describing test parameters and the procedure for dissolution testing analysis. The students in groups of two or three were assigned different experiments and asked to develop a method for their experiment following the recommendations given in the USP monographs and perform the method validation during one 3 or 4 h laboratory class period. The students were guided into the operation of all the specific equipment based on dissolution testing and UV or flame atomic absorption. Once these methods were developed they wrote complete SOPs for the validated analytical methods for dissolution and instrumental analysis.

n

f1 = ([ ∑ |R t − Tt |]/[ ∑ R t ]) × 100 t=1

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t=1

(1)

f2 = 50 × log([1 + (1/n)Σt = 1n(R t − Tt )2 ]−0.5 × 100) (2)

where n is the number of time points, Rt and Tt are the dissolution value of the reference and test batch at time point t, respectively. Profiles are not different when f1 is between 0 and 15. Two profiles are similar when the f 2 value is ≥ 50.27 Typically, up to 10% average difference in the profiles is assumed to reflect sameness in product performance in patients. Dissolution testing is therefore an important analytical tool in the development and manufacturing of pharmaceutical drugs. Since dissolution is rarely taught in undergraduate chemistry programs because dissolution testers’ are rarely found in undergraduate chemistry departments, B.S. graduates lack the fundamental concepts and essential skills for dissolution testing.

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EXPERIMENT ORIGINATING FROM KSU The experiment that was developed and validated was the “Analysis and Dissolution of Ibuprofen Tablets”. The test B

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Figure 1. External standard calibration curves for ibuprofen at 264 nm for each respective school.

Figure 2. External standard calibration curves for two groups of students from each respective school.

found under the dietary supplement section of the USP. The listed tolerance or Q value for Potassium Gluconate Tablets is that not less than 75% of the labeled amount of potassium gluconate (C6H11KO7) is dissolved in 45 min.32 The FAAS method was first validated using potassium standards prepared by dilution from a 1000 ppm reference standard for potassium, a potassium hollow cathode lamp and air/acetylene flame with detection at 766.5 nm as per the USP method.32 Cesium chloride was used as the ionization suppressor for the FAAS method. While the figures of merit were developed, only the test for tolerance (Q) was determined, not a dissolution curve. The students from SJFC wrote the complete SOP for the validated dissolution flame atomic absorption analysis method. The students at the other college received the method transfer with the associated pharmaceutical product in a different semester and performed their own validation following the SOP on a similar instrument. Any required deviations from the SOP needed to allow for the procedure to be completed were noted. The students compared their method validation to the one provided from the former school and drew conclusions based on statistics as to whether the method transferred successfully or not. These students then made any needed adjustments to the SOP and this experiment was transferred back to the originating school to see whether the next set of students can replicate the method validation following the SOP on the identical instrumentation as used before. Thus, each experiment went through two iterations of student AMT.

parameters and procedure for this analysis were obtained from the monograph for ibuprofen tablets found under the USP. According to the USP, samples labeled as “gelatin coated” are analyzed by UV at 266 nm minus the absorbance at 280 nm. The USP method lists a tolerance or Q value of not less than 80% of the labeled amount of ibuprofen (C13H18O2) is dissolved in 60 min.32 In the USP, the quantity Q is the amount of dissolved API specified in the individual monograph expressed as a percentage of the labeled content. This method was first validated for its figures of merit prior to dissolution testing using the USP Reference Standards for Ibuprofen. The dissolution procedure involves the drawing of samples from the dissolution medium during the test at several time points to create the curve. These samples are immediately filtered to avoid further dissolution by small insoluble particles sucked up into the syringe. Using six vessels and six time points, thirty-six samples are generated for analysis. The KSU students wrote the complete SOP for the validated analytical method for analysis and dissolution. This, along with sufficient samples, were sent to St. John Fisher College to see if they can replicate the figures of merit for the analysis and the dissolution profile as determined by f1 and f 2. Once replicated, the validated method with any corrections was sent back to KSU in order for another set of students to perform the method validation again.

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EXPERIMENT ORIGINATING FROM SJFC The experiment that was developed and validated was “Analysis and Dissolution of Potassium Gluconate Tablets”. The method of analysis is by Flame Atomic Absorption Spectrophotometry (FAAS). The test parameters and procedure for this analysis were obtained from the monograph for glucosamine tablets C

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RESULTS OF STUDENT VALIDATED UV−VIS SPECTROPHOTOMETRIC METHOD Ibuprofen standards were prepared in the range of 0−400 ppm. An external standard calibration method was used to validate the UV−vis method. The absorbance of all standards and a blank were measured five times at 264 nm on a UV−vis instrument. See Supporting Information for SOP received from KSU for preparation of ibuprofen standards. Calibration curves were created to demonstrate that the instrument response was linear, sensitive, and within the intended linear range of the expected concentration of ibuprofen in Advil, the tablet of interest. Linearity was determined by linear regression analysis of five ibuprofen standards within 80−120% of the nominal concentration. The method transfer acceptance criterion for linearity was an R2 > 0.995. The results for linear regression analysis of calibration data generated by students from KSU and SJFC (Figure 1) indicates that both schools met the acceptance criterion for linearity. The R2 value for the calibration curve generated by students from KSU was 0.9982 and that generated by the counterpart school (SJFC) after following the SOP received from KSU was 0.9991. Sensitivity a measure of how the signal changes as the amount of analyte is varied was also used to illustrate that the instrument response to ibuprofen met the intended purpose. It is noteworthy to mention that the sensitivity for the two calibration curves shown in Figure 1 turned out to be exactly the same (0.0017 absorbance units/ppm) despite the fact that the manufacturer and model of the UV−vis spectrophotometers used were different. In either case, the standard deviation of replicate absorbance measurements was very small such that the error bars of each graph are too small to be seen. A measure of the UV−vis method’s precision within each of the two schools was used to establish the reproducibility of the method. The results displayed in Figure 2 show that the calibration curves for each pair of students from each respective school had comparable precision. The intermediate precision between two analysts making use of the same equipment over a short interval of time was determined from the results of two groups of students in each respective school and using the paired student’s t test for comparison. It was determined that the results of the calibration methods were equivalent at the 95% confidence level with calculated t values of 0.458 (KSU) and 1.781 (SJFC). These calculated t values were found to be less than the tabulated t value of 2.78 at 4 degrees of freedom. The results of students validated flame atomic absorption methods for analysis of potassium gluconate tablets and the SOP received from SJFC can be found in the Supporting Information.

location from the vessels at predetermined intervals and immediately filtered to avoid further dissolution of the API. The filtered tablet samples were analyzed using a UV−vis instrument. The absorbance of all samples, one standard and a blank were measured five times at 264 nm. The concentration of dissolved API was determined from the application of Beer’s Law. The Q value was calculated from the concentration and the volume using eq 3: % Dissolved =

(SC (mg/L) × 100 (LC (mg/L)

(3)

where SC is the tablet sample concentration, and LC is the label claim concentration. The % dissolved value was compared to the expected Q value to pass the test. The dissolution test clearly passes the 80% Q value of the labeled amount dissolved in 60 min based on data obtained from both schools (Table 1).The standard deviation of the Table 1. Average Percent Dissolved Ibuprofen (Q Value) for a 1-Point Dissolution Test School

Time (min)

Q Value (%)

Kennesaw State University St. John Fisher College

60 60

88.3 ± 1.6 91.85 ± 4.93

average Q values of six vessels was better for KSU compared to SJFC, but the average Q value was within experimental error. Sampling four times throughout the dissolution test resulted in profiles that were similar over time (Figure 3). Calculations were made to allow for the loss of the volume taken at each time point. There was no significant difference observed in the dissolution profiles for Advil between the two schools and the dissolution results from both schools were within the experimental error.

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ASSESSMENTS OF LEARNING OUTCOMES

Both groups of students from SJFC and KSU were evaluated with similar expected student learning outcomes and assessment methods. Various quantitative and qualitative measures, including pre- and post-tests, and interviews were used to measure the findings from the evaluation activities. The learning outcomes were assessed using a standard pretest and post-test method through the use of both multiple choice and free-response questions. The pretest and post-test questions and full results are included in the Supporting Information. A nonparametric statistics approach was used to interpret the data generated from these tests so as to enable effective assessment of significant learning gains by students in respect to AMT.33 Interviews with students were conducted to determine if any learning gains can be attributed to the AMT conducted between SJFC and KSU. The interview protocol was semistructured.34 The interviews were conducted via Blackboard Collaborate, an online communication portal similar to Skype.35 Students were invited to participate in an interview using a stratified random sample. The interview protocol followed-up on questions raised by the pre- and post-test results and explored students’ perceptions and experiences of the lesson as well as factors that facilitate or inhibit their learning gains. Interview transcripts were analyzed qualitatively using analytic induction.36

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RESULTS OF STUDENT VALIDATED DISSOLUTION TESTING COUPLED TO UV−VIS DETECTION OF IBUPROFEN This example illustrates the validation of the Q value, which is the amount of drug product that needs to be dissolved in a set period of time during a dissolution test. Advil tablets in which the API of interest is ibuprofen were analyzed. Each Advil tablet is reported to contain 200 mg of ibuprofen and not less than 80% (Q value) of the labeled amount is dissolved in 60 min. The test was conducted at 37 °C using six round-bottom 1-L vessels, specifically designed paddles and a highly specialized sampling procedure. Advil tablets were placed in pH 7.2 phosphate buffer medium and samples withdrawn at a certain D

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Figure 3. Dissolution profiles of Advil tablets at 37 °C in pH 6.8 buffer (n = 6).

Specific student learning outcomes addressed from the AMT component conducted between SJFC and KSU showed that the students were able to • Define and understand figures of merit and decide which are important in method validation. • Define and understand what a validated method is versus a nonvalidated method. • Define and understand why method validation is performed as it applies to AMT. • Define the purpose of a dissolution testing procedure. • Develop a dissolution testing procedure and describe the expected results of a dissolution testing procedure. • Learn about and operate dissolution testers, UV, and flame atomic absorption (FAA) spectrophotometers. • Perform dissolution testing and UV and FAA method validation as it applies to AMT. • Calculate the validation parameters and equivalency from acquired data (precision, linearity, accuracy, ruggedness). • Write an SOP for the dissolution testing and UV or FAA assay of a pharmaceutical product (ibuprofen and potassium gluconate tablets). • Describe and understand what AMT involves and when it is performed. • Take part in assessing and making necessary changes to another lab’s SOP. • Understand what would make an AMT unsuccessful.

considerable increase in the number of students getting the correct answer going from pretest to post-test except for questions 1, 9, and 10 where only a slight increase or no change was observed. A slight decrease was observed for question 8, whereas a noticeable decrease was observed for question 4 (full results are included in the supporting information). This may be due to a plausible but incorrect answer among multiplechoice answers.

PRE- AND POST-TESTS The pretest was run in early September for both SJFC and KSU. The same assessment was given to the same students at both institutions in mid-December of the same year. A summary of the pretest and post-tests results evaluated using Wilcoxon signed ranks tests is shown in Table 2. This test is an alternative to the Paired Student’s t test, which is used when the population cannot be assumed to be distributed normally. The results of the Wilcoxon signed ranks test across all 3 years of the study indicated that students who took part in the AMT study at both SJFC and KSU had significant learning gains in all but two instances (p < 0.001). The exceptions were at SJFC in 2015 (p = 1.000) and KSU in 2014 (p = 0.105). The multiple choice questions and results analysis for the pretests and post-tests administered at SJFC and KSU showed a

INTERVIEWS Interview comments also showed how well the students were able to develop a procedure, write an SOP, and perform method validation and transfer. Table 3 contains the most pertinent student’s responses from the interviews. When reflecting on their experience, students in the analytical chemistry and instrumental analysis lab course at SJFC and the advanced analytical chemistry lab course at KSU provided some common themes: • Working in the laboratory while learning AMT was a positive experience. • Students gave an accurate description of AMT and what would make an AMT unsuccessful. • In regards to confidence with figures of merits the responses were mixed with some of students lacking confidence in their ability to fully understand them.

Table 2. Comparative Pre- and Post-test Results Evaluated with the Wilcoxon Signed Ranks Test Academic Year

School

Students, N

Median Scores,c Pretest

2012−2013

SJFCa KSUb SJFCa KSUb SJFCa KSUb

19 7 17 8 12 14

13 28 54 53 48 56

2013−2014 2014−2015

Median Scores,c Post-test

p Values

40 47 71 59d 54 69

0.001 0.018 0.002 0.105d 1.000 0.001

a St. John Fisher College. bKennesaw State University. cScores had a possible range of 0−100. dEight students who took part in the AMT at KSU were seniors who had already completed quantitative analytical chemistry and thus had good background on figures of merit before taking the pretest.

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Table 3. Typical Student Interview Comments by Learning Outcome Learning Outcome Laboratory Experience

Using the Dissolution tester

Learning AMT

Learning and Understanding Figures of Merit Understanding AMT

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Student Reflections “It felt like it had a purpose.” “It was really neat working in lab. I got to learn a lot of new stuff”. “It seemed like everyone worked well together.” “I thought it was helpful getting an SOP from another school and knowing it was written by someone our own age was really helpful.” “I thought it was a good experience you actually get to learn a lot of things···.” “The instructions for using the instrumentation were good. She walked us through it the first time.” “Being able to actually do it. Like performing an experiment using the dissolution tester because I mean in lecture it’s kind of hard to conceptualize what exactly was going on with all the different parts and things like that, but actually doing it, using it all, kind of brought it together as to this is how it works together and what each part does and when it’s done you know all that kind of stuff. It kind of gave it a tangible thing to go on.” “Having a USP Standard dissolution tester that is used in industry, being able to see it and learn the terms and how to run it and the process. I learned a great deal about it [dissolution testing] and still retained a great deal of the knowledge that I learned.” “I like the fact that I was doing something with AMT other schools looked at.” “I thought it was beneficial to the point that I can be let alone to a project and just working with somebody else’s SOP and just work off that and then build on that instead of the pre-cut kind of lab manuals where we were given the experiment.” “The assigned one [Figure of Merit] actually doing it made me remember it better.” “We worked a lot on it and we talked about it in class and we did a little worksheet about figures of merit and having that background going into the project I thought was helpful.” “The specific difference between each one [Figures of Merit]. Going through the Figures of Merit trying to specify which one is which. I had a little bit of trouble understanding which is which based on a certain area.” “If everybody who participated didn’t fully document what they did, if it (SOP) were transferred to another lab where they are going to try it ([SOP) they would not have an accurate description of what has been done and they would have to make changes to produce the same results.” “AMT is a procedure done in order to validate instructions so that you can use the same technique in every lab and you will still get the same results or nearly the same” “If you have a validated method comparing it from one lab to another and they do not correlate or correspond very well.” “Go back and look, see what parts weren’t successful, was it not robust, was it not specific, look at your figures of merit see where it failed and see if you can’t revamp it, come up with a new procedure or just change a part your procedure and see if that would help.”

• Students felt confident in their ability to use the dissolution tester. • Students were able to create an SOP for a pharmaceutical product and use the SOP (with 3 transfers) in the laboratory setting.

validation. These are guidelines which is something that we do every day in our job where cGMP guidelines are enforced by FDA and EPA”. • “My experience with the laboratory AMT was really positive. I really liked the opportunity to evaluate different SOPs and find mistakes in them that I might not have seen if I wrote them myself and it kind of gave me a different perspective of seeing the way that some people wrote things and the little directions that they missed out. Now that I work in industry, I see SOPs that we use on daily basis and sometimes my reaction is, oh! something is missing here, this does not make as much sense as it should. Having that opportunity to write our own SOPs and evaluate SOPs from the other school really helped with being able to take out those details that should have been included that sometimes always aren’t, and being able to understand them better and constantly update the SOPs to make more sense to anybody that reads them”. In describing job performance, results, and key accomplishments, one employer states • “The employee’s responsibilities comprise developing an SOP to test the presence of “chemical” in our finished products including validating and improving the current SOP used in our Testing lab and adapting it for use in the Materials Analysis Lab. She has developed a usable calibration curve after overcoming some unforeseen technical issues related to the calibration standards. In addition, she has found accurate alternatives to this technical hurdle and developed calibration curves for 20 separate standards with acceptable correlation values. On the basis of this results, she has written an SOP for testing the “chemical” in finished products and submitted it to the Lab Technicians for feedback in its usability. Her

FEEDBACK FROM B.S. GRADUATES AND EMPLOYER In describing how participating in this laboratory AMT experience has played a role in performing their duties in industrial laboratories, three B.S. graduates state • “Knowing the purpose and importance of SOPs is one of the things that helped me get my first job after graduation. This knowledge and experience automatically put me ahead on my training schedule. Since then, I have gained experience in pharmaceutical, medical device, cosmetic, and OTC testing/data analysis, in all of which following SOPs are extremely important. As a lab analyst, SOPs are required to be up to date and within reach for every step of any procedure (weighing, rounding, assays/ titrations, and instrumental analysis), and are one of the first things requested in an FDA audit. Outside of the lab, it is important to know how to write or change an SOP, or use them for creating protocols and reviewing data. Analytical method transfer is also applicable because currently, I work with third party vendors and testing sites to ensure testing is done correctly and per the method, and data is within the required specification. Additionally, many of the projects I am working on involve several groups of people from internal sites throughout the country”. • “The laboratory AMT experience really gave me good exposure to reading and understanding how SOPs work and learning to work with them during method F

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work has been instrumental in better understanding the chemical content of our finished products, and has been critical to the Company’s comments to European authorities on proposed legislation”.

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CONCLUSION Undergraduate students do not often have the ability to perform work that is done in industry and this experience enabled them to perform AMT as found in the real world. The students indicated that they liked and appreciated the laboratory experience of writing an SOP and conducting analytical method validation and transfer with students from another school and learning what goes into developing a successful AMT. Students had the opportunity to practice using a technique employed by B.S. level Chemists in the pharmaceutical industry. The students assumed the role of a Lab Manager and examining the data to determine whether the AMT was successful. The laboratory skills and hands-on experience that students have acquired from engaging in the laboratory AMT have enabled them to stand out in the job market and be recognized by employers.

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ASSOCIATED CONTENT

S Supporting Information *

SOPs from experiments originating at KSU and from SJFC as well as those containing changes that were made by students. The Supporting Information is available on the ACS Publications website at DOI: 10.1021/acs.jchemed.6b00878. Results of students validated flame atomic absorption spectrophotometric method and one point dissolution test for potassium gluconate tablets (PDF) SOP for preparation of ibuprofen standards at KSU (PDF) SOP for potassium standards preparation and dissolution testing; FAA analysis of potassium gluconate tablets (PDF) SOP for preparation of ibuprofen standards at SJFC (PDF) SOP for preparation of potassium standards (PDF) Multiple-choice questions and percent results observed for each question on pre-/post-test administered to assess learning gains in AMT (PDF)

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AUTHOR INFORMATION

Corresponding Author

*E-mail: [email protected]. ORCID

Irene Kimaru: 0000-0002-6277-5050 Notes

The authors declare no competing financial interest.

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ACKNOWLEDGMENTS We acknowledge the National Science Foundation (NSF) for financial support (Grant Nos. 1141021 and 1141042). We thank all the students in the analytical chemistry courses at SJFC and KSU during 2012, 2013, 2014, and 2015.

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REFERENCES

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