Sample Case Study on Good Laboratory Practice Principles

GLP study plan

Introduction

The good laboratory practice principles were issued by the FDA as mandatory regulatory requirements for laboratory practitioners on 29th June 1979. Even though the FDA has altered the contents of the regulations stipulated in the GLP guidelines, the scope has remained unaltered and still applies for non-clinical safety practices applied for drugs administration. The GLP principles apply in the entire OECD countries worldwide. Due to its sound applicability and a wide array of its address to pertinent matters in the fields of medicine and research, the principles have been adopted as the basic principles used in determining safe laboratory practices for the WHO/TDR. The GLP guidelines are a set of regulatory measures that can be divided into three main phases. The first phase of the guiding principles is the study plan whereby the practitioner has to put in place proper planning mechanisms that would be followed while carrying out the experimentation. The second phase is the experimentation stage whereby the actual experimentation process is conducted. The experimentation phase is carried out according to the plans laid down in the planning stage. The last phase described in the GLP principles is the reporting phase. It is at this stage that the practitioner gives an elaborate report about the findings derived from the experiment.

The study plan stipulates all the requirements as well as step by step instructions that are to be followed while carrying out the experimentation plan. Besides, the plan also describes the reporting mechanisms that would be used to deliver the findings in a manner that can be best understood by all members of the general public. Good Laboratory Practices have been widely applied in conducting laboratory experiments with an aim of ensuring better the generation of better and well informed scientific results for sound decision making. Most OECD nations have fully ratified the specifications described in the OECD’s good laboratory practice principles as a standard measure for conducting valid scientific researches and experimentations. Scientific experiments are conducted under controlled environments treated in different procedures in order to guarantee uniformity of the results as well as allow sound judgments for scientific discoveries. For an effective experiment, all the three phases involved in the experimentation process as described in the OECD guidelines must be taken into consideration. Scientific procedures and conducted in a stepwise mode whereby each step is accomplished at a specific time. The succeeding steps are determined by the success in the preceding ones. Illustrated in this document is case scenario of a rat GLP study plan conducted at a facility in my work station. The study plan has been developed in line with the OECD guidelines 407 describing the fundamental principles that underlie the GLP practice among the member countries. I have described this study plan using a case scenario developed by the facility.

Title of study

The title of this study is informed by the anomalies registered in our random observations and which call for effective address.

Toxicokinetic analysis of plasma compounds (100mg/kg) in in-vitro cultured mammalian (rat) cells using Lithium Heparin in an LAF- 001 test method

Objective

After the completion of this study, we aim at achieving the following objective

To successfully conduct a toxicokinetic analysis of plasma compounds (100mg/kg) in in-vitro cultured mammalian (rat) cells using Lithium Heparin in an LAF- 001 test method.

Nature and purpose of the study

The study will be conducted in in-vitro condition and the results generated examined within the context of a natural environment. Several results generated from a series of experiments will be compared to create harmony for better-informed decision making. Relevant chemicals Lithium Heparin and HO2 will be applied under regulated conditions for the best results. This is aimed at eliminating biases and environmental disruptions. At the end of this study, we purpose to determine the level of toxicity in the rat plasma by applying the efficacious dosage of powdered water at interval rates within standardized retest duration. The study is expected to be informative in nature and inform decision making to steer the discovery of new insights as well as the validation of set hypotheses. It is based on the intended objectives that the study is designed after a careful analysis of observed anomalies.

Identification of the study, test substance and reference substance

Reference substances to be used

Product line                                        Lithium Heparin

Catalog number                                  1234

Size                                                     56mg

Surface chemistry                               98% pure HO2

Solid content                                       700 mg/kg

Quantity                                              100mg

Media composition                             HO2, (distilled)

Buffer, preservatives                          none

Particle shape                                      spherical

Particles density                                 2.3g/cm3

Particles porosity                                porous

Particles stability- stable in                HO2, (distilled), aqueous buffers

Product form                                       aqueous solution

Volume of one particle                       not measured

Mass of one particle                           not measured

Surface area of one particle                not measured

Number of particles/ ml of solution   not measured

Binding capacity                                not available

Charge density                                    2.1mg/m3

PH                                                       6.8

To be stored below -200C- Stable in the vehicle for 7 days at 2-80C

Information on the sponsor and test facility

The experiment is designed and sponsored by qualified personnel in line with the requirements of the OECD on good laboratory practice. The qualifications of the participants are described as shown in the succeeding sections of this plan. Besides, well equipped and maintained laboratories have been selected to aid in carrying out the experiment as per the specifications of the OECD. The facilities that will be used for conducting the experiment, observations and analysis of the results generated have also been selected based on strict regulations described in the OECD guideline 401. These specifications have been described in the succeeding sections below.

Sponsor: Dr Craig Smith of New Technologies Ltd, 300 Meyo Street, Camptom, NSW, 3402, Australia; c.smith@NT.com.

Principal investigator: Dr Simon Alve- sample analysis

Test facilities and test sites involved: St John’s mission hospital, Prime Laboratories, ADME Pty Ltd, Bioanalysis St, Colorado, 11573, USA

Dates of study plan                                                                             Signature

Event                                                   Date                             Sponsor             Management

Commencement of experiment          01/06/2015                  ………….                   ………….

Completion of experiment                  01/10/2015                  ………….                   ……………

Test facilities

St John’s mission hospital, Colorado, 24365, USA

Prime Laboratories- toxicokinetic analysis, Colorado, 41463, USA

ADME Pty Ltd, Bioanalysis St, Colorado, 11573, USA

Test method description

The test shall be conducted in reference to the OECD test guidelines. The international standards will be taken into consideration to make sure that all guidelines are met to success. Specifically, the study will be conducted in accordance with the OECD guideline 407. The efficacious dose of the compound that will be used is 100mg/kg. The solubility test item that shall be used, LAF- 001 shall limit the highest dose that shall be assessed at 700mg/kg. The test shall be conducted using a predesigned vehicle in order to ensure that the reagents work in suitable conditions that promote the generation of accurate results. In accordance to this aspect, the vehicle that has been selected for this test is water. The test item, Lithium heparin, is stable in the vehicle used for a period of seven days when kept at temperatures between 20C and 80C. Such conditions will be ensured by regulating the temperatures under which the test item will be stored. In this regard, the test item will be put under appropriate conditions to ensure the sustainability of the experiment and the validity of the results generated. The test item will be sued in its powdered form and purity maintained at 98%. The item, Lithium Heparin will be stored at temperature below -200C to keep it at stable condition. This will prevent contamination and maintenance of its chemical component. The powder is moderately reactant with other chemicals such as moisture, water and other agents floating/ suspended freely in the atmosphere hence leading to inappropriate results. The test item, according to the required standard specifications, the retest date is pegged at October 2015.

After the completion of the retests, a toxicokinetic analysis of plasma samples is to be performed on the last day of dosing. In order to maintain the consistency of the results for well-informed decision making, the time points for intravenous blood collection will be designated at pre-dose, 10, 30, 60 min, 4, 8, 12 & 24h post-dose. Blood samples will then be collected into tubes containing Lithium Heparin. The collected samples will then be centrifuged within 30 minutes of collection to maintain the quality of the results required and inform adequate decision making. In order to ensure the validity of the results generated, the resulting plasma will be divided into two aliquots and with the primary samples stored at -80°C prior to shipment. These samples will be suitable enough for shipping since all repercussions shall have been taken into close considerations. A bioanalytical method validation for LAF-001 in rat plasma has been undertaken at ADME Pty Ltd, Bioanalysis St, Colorado, 11573, USA and the samples are to be transferred there (on dry ice) for analysis. The person that will be responsible for sample analysis is Dr Simon Alve.

Justification for selection of the test system

The OECD GLP principles require that the test materials be kept safe from any form of contamination in order to arrive at the best results intended. In regard to these requisitions, the vehicular solvent will be kept in places where they are free from contaminations and reagents that may cause impurities and effect results. In order to ensure the attainment of these required standards, distilled water, that is free from impurities, will be used to eliminate inconsistencies and influences on the intended results. The test items will be kept under standard conditions e.g. Lithium Heparin stored at temperatures maintained at -200C in order to ensure the attainment of the best results. Besides, distilled water, free from any form of contamination will be used in order to ensure the validity of the results by reacting pure reagents. All the apparatus and chemicals that shall be used in the experiment will be sterilized and kept free from contamination. Such contaminations can influence the test results leading to unintended conclusions and interpretations. This will consequently lead to wrong conclusions hence invalidity of the experiment. The OECD guidelines advocate utter care to be taken by practitioners to eliminate inconsistencies resulting from contaminations. Relevant measures will for this reason be put in place to ensure that the results generated are sound and applicable to the entire scientific community as well as inform sound decision making on related issues. The OECD guidelines also advocate the participation of excellently trained personnel. These persons must be able to adhere to the strict guidelines of the Good Laboratory Practice principles as stipulated in the OECD guidelines. It is for this reason that highly trained and reputable personnel were selected to conduct the experiment in order to generate sound results. Definite stages will be carried out by specialized personnel in order to ensure the reliability of the results generated.

Test substances

Identification                                                  Registration

Lithium Heparin                                             L13H23

HO2                                                                 A, HO2, 32

Safety precaution

Safe storage of chemicals will be guaranteed by taking relevant measures to ensure that the items used in the experiment are free from contamination as well as avoid accidents. Among the precautionary measures that shall be taken are described in this section. All chemicals will be tightly sealed to avoid spillage and accidents. This measure will also prevent the chemicals from contamination through reaction with other reagents. All apparatus that will be used during the experiment will be disinfected to avoid interference with the results generated. This will also guarantee the validity of the results generated.

Conditions of exposure

Lithium Heparin- Stored at -200C

– Kept in stable conditions for 7 days at temperatures between 2- 80C

Distilled HO2      – kept in closed, sterilized and disinfected container

Dose levels, frequency and duration of exposure

The dose for the test item will be assessed to 700mg/kg as per the requirements while using the LAF- 001. The duration of exposure is designed to take 4 months. Intravenous blood collection shall be conducted at the frequency described as follows: pre-dose, 10, 30, 60 min, 4, 8, 12 & 24h post-dose

Type and frequency of analysis, measurements, observations and examinations to be performed

The level of toxicity in the plasma content of the rat shall be determined. This shall be realized by observing the colour changes in the mammal’s plasma. Changes in PH values of the plasma content will be determined after the addition each dose of lithium heparin. The PH value will be determined with the aid of the PH scale depending on the color changes observed. The changes in plasma content will be used as a measure for toxicity levels. Measurements and observations of the expected changes will be conducted after each alteration and results recorded instantly. Changes in PH values will also be observed and determined by observing the changes on litmus paper. The changes will then be compared with the observations derived from the PH scales.

Statistical methods

A wide array of statistical methods will be used to analyze the data generated after conducting the experiment. A wide range of statistical methods can be used to simplify and make sense of the generated data. However, the best methodology is the one that will generate results that can be easily understood and interpreted with less efforts and little statistical knowledge. The ease of interpreting and understanding the results informed the selection of the methods chosen for this experiment. Harmonization of the results will be done by using the regression analysis. Analysis of variance will be used to determine the extent of variation in the toxicity levels using a confidence level of 99%. As opposed to descriptive statistical analysis where the confidence level need not to be very large, medical experiments need to be more accurate in order to guarantee the effectiveness of the results generated. It is for this reason that a confidence level of 99%, 1% error, was chosen for this experiment in order to ensure the accuracy of the results generated. Other descriptive statistical methods such as range mean and medium will also be used to effectively describe the results generated in the easiest way possible and make the experiment useful to the broader community. Data presentation will be done using a wide range of statistical methods. The changes in PH values in the blood plasma will be presented using line graphs. This is because the line graphs give a clear visual interpretation at a glance.

Interpretations of results

Interpretation of the results generated during the experimentation will be conducted in different ways. Written explanations will be provided each explaining the data in the statistical methods used in the presentation. This will make it easy to understand the results generated and presented in from the findings.

Standard operating procedures (SOP) concerning the study

The prime operating standard that will be used to guide all operations conducted during the study will be pegged upon the stipulations described in the OECD good Laboratory Practice principles. All standard specifications will be strictly followed while conducting the experiment. The generation of the results will as well be carried out in a following the strict procedures described in the OECD guidelines. The OECD guidelines 401 will largely inform the practical experiments conducted here.

Recording

A number of materials will be recorded and retained in the archives for future references. Among the records that will be archived include the study plan, amendments and deviations, draft reports, copies of the final report and other attached documents related to the study. In addition to the aforementioned reports, other records that shall be archived include the original raw data collected during the experiment and copies of the product registration forms to authenticate the results and the experiment at large.

Conclusion

To conclude, experiment shall be conducted under controlled conditions in order to generate the required results. Strict observation of the underlying principles stipulated in the OECD good laboratory practices will be ensured to ensure that the experiment meets the set standards set in the guidelines. Specifically, the OECD guidelines 401 will be used to inform the entire experiment. Proper and scientifically sound precautionary measures will be adhered to in order to minimize improper handling of the apparatus and chemicals. Observation of the precautionary measures will also help minimize accidents and alteration of the expected results and wrong conclusions. A wide array of statistical methods will be used to analyze the data as well as present and interpret the results. Regression and analysis of variance will be used to analyze the data collected at a confidence level of 99% in order to increase accuracy. Histograms and scatter diagrams will be used to present the results generated du