Inserting Sample on NMR Magnet

Inserting Sample  

1. NMR tube containing sample should be held in a plastic spinner (use the blue color spinner in 500 and 600 MHz spectrometers; use the white color spinner in 700 MHz spectrometer).  Hold the sample by the top, place sample tube in the spinner and the spinner in the sample depth gauge. Push or pull the sample tube so that the depth of the sample above and below the center line of the sample depth gauge is equal. However, never exceed the lower limit (position of the adjustable white platform which should be at the line marked 5 mm ­  15mm) as this can damage the probe as well as the sample, as the sample will rest on the tip of the tube, not the spinner. This is important for all probes, but especially the cryoprobes installed on the 600 and 700 MHz spectrometers.

2. Remove the black cap from the top of the magnet bore. Next, press the LIFT button in upper left portion of the BSMS keyboard. Wait for the airflow (hissing sound that can be heard). Then remove the depth gauge before inserting the sample and spinner into the magnet. Pressing the LIFT button will toggle the air flow off and will drop the sample tube gently to the magnet bore where it will be positioned at the top of the probe.

Preparation of NMR Buffers Protocol

3. Preparation of NMR Buffers Protocol:

3.1.Stock Solutions:

3.1.1  0.1M stock solution of Na2HPO4.

        Weigh 141.96g of Na2HPO4 (anhydrous) and dissolve to 1000 ml (one Liter)     with ddH2O.

3.1.2  0.1M stock solution of NaH2PO4.

           Weigh 119.98g of NaH2PO4 (anhydrous) and dissolve to 1000 ml (one Liter) with ddH2O.

NB: The weight of each salt needed to prepare one liter of a 1M solution varies, depending on the hydrated state of the salt.

3.1.3    .50m M stock solution of DSS.

           Weigh 1.0915g of DSS and dissolve to 100 ml with ddH2O.

3.1.4. NaOH solution.

3.1.5. HCl solution.

3.2. NMR Buffer number 1:

3.2.1. Measure 28.85 ml of the 1M stock solution of Na2HPO4 and 21.15 ml of the      1M stock solution of NaH2PO4 into a 1000 ml graduated cylinder.

3.2.2. Mix and add ddH2O to 850 ml.

3.2.3. Add 100 ml of D2O.

3.2.4. Add 10 ml of the 50 mM DSS stock solution.

3.2.5. Mix well and adjust to pH to 7.0 (if necessary) with HCL or NaOH solution.

3.2.6. Make volume up to 1000 ml with more ddH2O

3.2.7. Store in a labeled, dated bottle in the refrigerator.

3.2.8. Check for impurities before use.

3.3.NMR Buffer number 2:

3.3.1. Measure 28.85 ml of the 1M stock solution of Na2HPO4 and 21.15 ml of the    1M stock solution of NaH2PO4 into a 1000 ml graduated cylinder.

3.3.2. Mix and add ddH2O to 980 ml.

3.3.3. Mix well and adjust to pH to 7.0 (if necessary) with HCL or NaOH solution.

3.3.4. Make volume up to 1000 ml with more ddH2O

3.3.5. Store in a labeled, dated bottle in the refrigerator.

3.3.6.  Check for impurities before use.

Probe Tuning :NMR

Probe Tuning :NMR

Probe tuning must be matched to the sample in order to minimize pulse widths and maximize sensitivity.  Although the effects are small for routine proton spectra, they can be dramatic for more complex pulse sequences.  Although the tuning changes between different organic solvents usually do not justify the effort to tune, if you are changing from an organic solvent to water or D₂O, retuning the probe is usually worthwhile.  If you are starting a long experiment, you will want to check the probe tuning.

The wobb command allows you to tune and match a probe in an easy way even when the coil is heavily mistuned and mismatched. First copy the parameters for the nucleus you wish to observe.

Now enter the acquisition window by clicking Acquire/Observe fid window and start the wobble routine by clicking on Acquire/Acquisition parameter setup/Tune probehead. Alternatively enter acqu and then wobb via the keyboard. The acquisition is started and after a few seconds the wobble curve is displayed and refreshed continuously. A vertical line is drawn at the center frequency

(SFOx) to provide optical information on the frequency which is to be tuned. The horizontal axis of the coordinate system is scaled in MHz and labeled accordingly. Useful information like nucleus, tuning frequency, frequency of the minimum of the wobble curve and wobble width, is displayed in the information window. Simultaneously the LED display on the preamplifier is set and refreshed accordingly. The wobble curve shows a dip downwards which changes while you turn the tune or match knobs of the probe.  Example displays in Figure. 

1. Turn the tune knob (tune sliders on the indirect probe) so that the dip moves towards the center of the screen. Keep on turning until the dip is exactly in the center across the vertical line.

2. Turn the match knob (match sliders on the indirect probe) so that the dip becomes deeper. Keep on turning until the base of the dip is at a minimum. This occurs at the zero level line for most probes.

3. On most probes the matching influences the tuning and vice versa, so repeat steps 1 and 2 until the dip is exactly in the center of the screen and its base at minimum level. Then the probe is tuned and matched.

When you are finished with tuning and matching stop wobb by activating the stop button or by entering stop via the keyboard.

Operating procedure for NMR spectrometer : Bruker Avance DPX-300 NMR spectrometer

Note:

Unless otherwise specified, statements such as “click on calib” mean to move the cursor on top of the calib button on the screen, and click with the left-hand mouse button. Consider the left-hand mouse button the default; if another button is required, it will be explicitly stated. 

Commands such as “Enter zg” mean to type zg followed by a return on the

keyboard. Commands must be followed by <return> before they are accepted.

Words written in bold Arial font, e.g., acqu, refer to buttons or pull down menus on the screen (items that can be selected with the mouse).  Multilevel menu choices written like this, Windows/lock, mean choose the first menu choice by left clicking with the mouse.  Choose the second menu choice from the menu that appears when the first choice is made.

Words written in bold Courier font, e.g., edsp, refer to commands that can be typed at the keyboard.  Commands must be followed by <return> before they are accepted.

General Procedures For All Spectra

 1.  Preliminary Notes

A.     This manual is a condensed and incomplete account of instrument usage.  It is in no way a substitute for reading the detailed instruction manuals available in the NMR room (M-02).  Please get to know them. This manual is written in a similar format to the Tufts AM-300 manual, to facilitate the transition from the AM to the DPX.

B.      All users should sign the logbook before they do anything else.

C.      Log into the computer.  The screen may be illuminated by hitting any key on the keyboard.  You should see a Welcome to IRIS window.  Enter your user name and password.  When the UNIX shell window appears, type xwinnmr.  The XWIN-NMR window will appear, as shown below.

D.  Only the active window will accept typing.  To make window active, position the mouse cursor inside it.  To bring a window to the front, left click in its title bar.

E.  Most commands are accessible from the menu bar.  Alternatively, any command may be typed into the command line below the spectrum window.  Most commands are the same as the AM-300.

F.  Parameters may be changed by typing the abbreviation for that parameter followed by the (return) key (hereafter <rtn>.  The computer will respond by popping a window with the current value of that parameter.  Typing the new value of the parameter followed <rtn>by will replace the old value with the new value.  Alternatively, groups of parameters can be edited with various ed commands.  For example, eda presents a window of all acquisition parameters, edp presents a window of all processing parameters.

G.  Sample control is provided through the BSMS (Bruker Smart Magnet System) control panel.  To access the BSMS Panel choose Windows/BSMS Panel from the menu. Lock, Shim, and Sample panels are similar in appearance to the hardware buttons on the AM-300, except control is through a slider bar and +/- button, rather than through the knob.  For all buttons the left mouse button decreases the value, the middle button increases.  Step size is changed by left (decrease) or right (increase)clicking on the :2/*2 button

2.  Loading the Sample

 A.  The probe should contain a sample at all times.  When you arrive at the instrument with your research sample, you should remove the standard sample from the magnet and replace it with your sample as follows:

From the XWIN-NMR menu choose Windows/BSMS panel.  Then click on the Shim button

-Click on the LOCK button to unlock the instrument.  The color of the button will change from yellow to gray

- Click on the SPIN button to stop the spinning of the sample The color of the button will change from yellow to gray

- Click on the  LIFT button.  The standard sample should rise out of the probe on a stream of air. Remove the standard sample from the top of the probe. Remove the standard from the spinner.

-Place your sample tube in the appropriately sized spinner and adjust the precise depth of the tube within the spinner by using the depth gauge.  Note that the depth gauge should be set at 10mm regardless of the tube size for a 10 mm probe. (and at 5 mm gauge for a 5 mm probe).  Note: 10 mm tubes cannot be run in a  5 mm probe. For best shimming set tube 1mm below the appropriate line to assure a uniform sample in the active region of the probe.

- place your sample in the top of the probe. Do not release the sample unless you are sure it is supported on the air current.

- Click on the LIFT button again.  Your sample should slowly lower into the tube as the air stream decreases. Click on the SPIN button.

B.  The rate of the sample spinning should be 20-25 rps.  To check this:

-Press the Sample  button on the BSMS panel.  Then click on the SPIN MEAS button on the Sample panel that pops up.  To change the spin rate, click on the SPIN RATE button, and usethe slider bar to change the value.

3.  Locking the Instrument

 A.  To display a swept lock signal, choose Windows/lock from the XWIN-NMR menu. You should now see a continuous wave deuterium NMR spectrum of your solvent.  This should be a trace across the screen with the signal in the middle of it.

B.  To lock the sample, type lock, then click on the solvent from the list that pops up.  Lock power, phase, and gain will be adjusted automatically.

C.  The lock can also be adjusted manually, using the buttons on the Lock panel from the BSMS panel.   Note: Appropriate power and gain levels are different from the AM-300.

 4.  Shimming the Magnet

 A.  To obtain a high resolution spectrum it is essential that all parts of the sample are exposed to the same magnetic field.  The homogeneity of the field is generally assessed by noting the intensity of the lock signal, the more intense the signal the better the field.  In practice, the shims most likely to require adjustment are the spinning ones, namely Z Z² and Z³ shims, and the non-spinning shims X and Y.  To adjust the homogeneity by hand (often adequate for routine spectra and almost always faster than by automated computer shimming) you must first make sure that the lock trace is visible.  If it has gone off the screen during the locking procedure it can be brought back by lowering the LOCK GAIN level.  A sample procedure follows:

- Click on the LOCK GAIN button. Use the +/- button to adjust the lock gain until the trace is in the top two squares of the grid. Press the Z button. Use the +/- button to maximize the lock signal.  Change the step size if needed.  Repeat for the Z² button. Continue to alternate between the these buttons maximizing the lock signal each time until no further improvement can be seen. You may have to adjust the Lock Gain if the trace goes off the screen.

-If the sample must be shimmed to highest standards, adjust the non-spinning shims by pressing the SPIN button and waiting until the sample stops spinning. Spin rate can be used to monitor this. Raise the lock level by adjusting the lock  gain until the level is in the upper half of the screen. Now press the X button and use the +/- button to adjust the lock level to maximum. Then press the Y button and repeat this procedure until no further improvement is possible. Start the sample spinning again by pressing the SPIN button and repeat the Z and Z² shimming as above.

Running a ¹H Spectrum

 5.  Setting the Parameters

 A.  From the XWIN-NMR menu choose File/New. A edc window will appear containing the file name parameters.  Enter a new NAME. Generally only the Name parameter needs to be changed. NOTE:  The first time you use XWIN-NMR you will need to change other parameters to EXPNO=1, PROCNO=1, DU=datanmr, User=<username>. Click on SAVE

B.  Data files containing typical parameters for a proton spectrum are stored in the hard disk for easy access.  To retrieve these:

Choose File/Copy/parameters from file from the menu, or type rpar. A list of standard parameter files will appear.  Use the scroll bar to access the entire list.  Click on proton. In the next window that pops up, click on Copy All.

Type eda to edit acquisition parameters.  Always change Prosol to True by clicking on its value, which loads all default pulse lengths and also check that the solvent is correctly identified, otherwise calibration will be off. IMPORTANT:  Failure to set Prosol to True will result in all pulse lengths not set by you will be of zero length and rga will go off scale trying to find a signal.

Change any other acquisition parameters desired, then click on SAVE. Acquisition,  Ones you might want to change are

TD (time domain size)-Defines the number of data points of the fid.

NS (number of scans)

P1 (pulse length in microseconds)

SW, SWH (sweep width in ppm, Hz)

AQ (acquisition time in seconds)

RG (receiver gain)

B.  You may wish to tune the probe before beginning acquisition.  You can see this on another post Probe tuning.

C.  The last parameter to be set is the receiver gain (rg) which adjusts the gain of the signal coming out of the probe.  This will vary from sample to sample.  To set this gain type: rga (receiver gain adjust). The computer will automatically take a few sample pulses and alter the RG between each.  When the best value is found, the computer will type rga: finished.  You may see this value which the computer has selected by typing: rg. If you want this value then type: <rtn>.  If you wish to select another number then type in the new number  and then type <rtn>. Do not increase the value, but you may decrease it.

6.  Acquiring the Spectrum

 A.  To start the acquisition type zg to zero the memory and go (start the acquisition).  You can watch the progress of the acquisition by choosing Acquire/observe fid window from the menu, or by typing acqu.

B.  The acquisition will automatically end when the number of scans taken equals ns.  However if you wish to stop it prematurely or abort the run then you will have to click on the STOP button to the left of the FID in the acquisition window.

7.  Transforming the Data

 A. A complete description of processing parameters and their meaning can be found in the  XWIN-NMR Software 

B.  The time domain data can be processed by typing bc, then em, then ft (or type ef to do all three at once).

C.  It is not necessary to wait for the acquisition to end before transforming the data.  Type tr to transfer the data to disk at any time during the acquisition.  Once written to disk, the data can be transformed.  Time domain (fid) and frequency domain (spectrum) data are always stored in separate files. All data acquired so for can be processed while the acquisition continues in the background.

8.  Manipulating the spectrum

 A.  The spectrum can be expanded vertically and horizontally using the mouse, clicking on the buttons to the left of the spectrum.  *2, /2, *8, /8 multiply or divide the vertical scale by 2 or 8.  To expand the horizontal scale, left click anywhere in the spectral window. Then middle click on the desired expansion limits.  Left click again to unlock the mouse from the expansion.  To expand the vertical scale by an arbitrary amount, left click and hold the double arrow button to the right of /8.  Move the mouse vertically until the peaks are desired height.  Release the mouse button.  

 9.  Phasing the Spectrum

 A.  The spectrum will usually not have all the signals upright in an absorption phase. To correct this type apk (automatic phase correction).  For spectra with well separated narrow lines, apk will usually work well.

B.  If the automatic phase correction does not work well, click on the phase button to the left of the spectrum, then click on biggest.  A marker will appear under the biggest peak in the spectrum. Left click and hold the PH0 button.  Move the mouse vertically until the largest peak is phased correctly.  Release the mouse button and click and hold the PH1 button. .  Move the mouse vertically until the other peaks are phased correctly.  Release the mouse button.

 10.  Setting the Reference

 A.  The spectrometer cannot provide an exact chemical shift without being calibrated to a standard (usually TMS).  Click on the calibrate button to the left of the spectrum.  A small arrow will appear on the spectrum.  move the arrow to the reference peak with the mouse, and click the middle mouse button.  A window will appear with the current frequency value.  Enter the reference frequency. A chart of chemical shift values for solvents is on the console.

11.  Changing SW and O1 for Increased Resolution

The default parameters display the spectral range 16 to –4 ppm.  This range can be changed for greater resolution.  Click the left mouse button somewhere in the spectral window to tie the cursor to the spectrum. Position the cursor at the left limit of the desired spectral width. Click the middle mouse button to set a marker at this frequency. Move the cursor to the desired right hand limit and click the middle mouse button to expand the spectrum. The expanded region now appears in the window. Click the left mouse button to release the cursor from the spectrum. Click on sw-sfo1 while the expanded region is displayed. This adjusts sw so that it has the same value as the expanded region and also adjusts o1 (and thus sfo1) so that the carrier frequency lies in the center of the expanded region. (You can verify these changes by checking the eda table.) Notice that by reducing the spectral width, the acquisition time aq is increased while the parameter fidres is reduced. Finally, now that the acquisition parameters are optimized, it is a good idea to repeat the automatic receiver gain adjustment (rga).  Notice that since the spectral width has been changed, it may be necessary to readjust the phase correction.

 12.  Integrating the Spectrum

 A.  It is often useful to integrate ¹H spectra. The simplest way to define the integral range is by entering abs. The command abs performs an automatic baseline correction and also automatically defines the integral ranges.  The integrals will not appear on the screen, but will appear on the plot.

B.  To display the integral on the screen:

Choose Analysis/Manual integration from the menu, or simply click on the integrate button of the button panel at the left side of the XWIN-NMR window.  Additional buttons, like those that control vertical and horizontal scale for the spectrum, will appear in the lower left.  The upper part of the button panel is identical to the standard layout, and allows you to shift and scale the integral data on screen. In addition, there are three special sections headed by current:, all:, and mouse:. The command buttons in these sections work on the current integral marked by the user, on all integrals on the screen, and on the mouse sensitivity, respectively.

Defining integration regions

Move the cursor into the data area of the XWIN-NMR window. Click the left mouse button. The cursor is now bound to the spectrum, and moves along the spectrum trace when you move the mouse. It can be released from there by clicking the left button again. Clicking the middle button will mark the current position (the mark can be removed using the right button). Clicking the middle button a second time at a different cursor position will define the area between the mark and the current cursor position as the integration region, and the corresponding integral trace is displayed along with the value of the area under the integral. By default, the first integral region defined will be assigned the value 1.0. This procedure can be continued for all desired regions, and need not proceed left to right. Click the left button to release the cursor from the spectrum when you are finished.

In order to mark one of the defined integrals as current integral, move the cursor into the data area of the XWIN-NMR window. Click the left mouse button. The cursor is now bound to the spectrum, and moves along the spectrum trace when you move the mouse. Select the integral you want to make the current integral by moving the cursor under it, and release the cursor by clicking the left button again. The integral will be marked with an asterisk. All button panel commands in the section current: can now be applied to this (and only this) integral.

To calibrate the integrals, mark an integral as the current one, then click on calibrate.  Enter the integral value in the dialog box.

When all integral regions have been defined, click on return. Click on save as ‘intrng’ and return to save the regions to a disk file.

13.  Peak Picking

The most straightforward way to produce a peak list is through the command line.  First display the Y axis in cm, if it is not already displayed.  The Y button to the left of the spectrum window will toggle the y-axis, the YU button will change the units from cm to absolute.   Choose the region for peak picking by setting the parameters f1p and f2p to the ppm values for the left and right limit, respectively. (If the x-axis is in Hz, use f1, f2 instead of f1p, f2p.) Set the parameter mi to the desired minimum intensity in cm.  Type edo and check that CURPRIN is set to hplj5l. The command pp will produce a peak listing on the printer.  Use pps to print to the screen.

14. Plotting the Spectrum

A straightforward way to plot 1D spectra is by using most of the plotting parameters found in the plot parameter file standard1D. Read in the file by choosing File/Copy/parameters from file from the menu, selecting standard1D from the menu of parameter file names, and then selecting plot from the menu of parameter file types that appears.  Then click on Copy. This sets the plotting parameters to values appropriate for most 1D spectra.  1D and 2D Plotting Parameters’. For basic 1D spectra no changes need to be made within the parameter menu edg itself; however, the spectral region and the integral range must be defined, and the spectrum title must be written. To select the spectral region (full or expanded) to be plotted, first make sure the spectrum appears as desired on the screen, and then click DP1and simply hit return in response to the following three (3) questions:

F1 = <return>

F2 = <return>

Change y-scaling on display according to PSCAL?<return>

You may change the values of F1 and F2 if you wish.

Other plot parameters can be changed by typing edg (edit graphics) and changing the values, mostly yes/no, which determine whether integrals, parameters, etc appear on the plot. The edg window is analogous to the DPO command on the AM-300. Some plot features have a button labeled ed.  Clicking on this button will open another window, with parameters that control the position and style of these features.  If simply editing the yes/no parameters in the edg dialog box will not give the plot you want, it is usually easier to use XWIN-PLOT, rather than to change other plot parameters.  XWIN-PLOT is a graphical plot layout program.

Next create a title for the spectrum. Enter setti to use the editor to open the title file. Write a title and save the file.  The title must end with a <return>.

To preview the plot, as it will appear on paper, type view.

To plot the spectrum, type plot (provided the correct plotter is selected in

edo).

Is UHPLC Worth All The Hype

Is UHPLC Worth All The Hype

UHPLC or Ultra high performance liquid chromatography can be a technique, or sort of chromatography that’s differentiated from Non HPLC from the method which the separation takes place. The HPLC or UHPLC takes place by using a pump that transfers the substance through the mobile phase as well as the column along with a different sort of detector. Shorter columns are possible for the reason that columns are far more densely packed. Gravity can’t be used again because of the density of the column material.
High performance Liquid Chromatography is used within the laboratory to be able to separate one substance from another and also to analyse what that substance may be. This is achieved for medical, for business, or for law enforcement procedures so just about any type of business today might use UHPLC.
The detection method currently in use signifies that a specific amount of retention time is needed and understanding that the pump instead of gravity can be used to maneuver the analyte. Regular column chromatography often requires a longer column in comparison with the HPLC since particles in HPLC are much smaller and much more densely packed, permitting a better look considering that the separation is far better.
Normal chromatography effects the separation from the analytes determined by their absorption from the analyte by a stationary surface. HPLC or UHPLC offers a much increased pressure, which is necessary in order to get the mobile phase as well as the analyte with the columns. The pumps will be different inside the pressure that they offer but they are measured through the ability they must obtain a consistent flow rate and to provide a consistent pressure for that movement from the substance to get analyzed.

Preparation Of Sampling Bottles

             Preparation of Bottles for sampling:

·        The amber coloured bottles used for sampling shall be issued from packaging stores.

·        Wash the bottles to be used for sampling thoroughly with plenty of purified water.

·        Dry these washed bottles at 105° C for 2 Hrs.

·        Close the bottles and keep these bottles in a polybag and cable tie. Use these bottles for sampling purpose.

·        The cleaned bottles shall be used for sampling within three days after cleaning.

·        After 3 days the balance bottles shall be discarded.

The reconciliation of sampling bottles shall be carried out and the same shall be recorded in the Usage log book of Sampling Bottles

Research and Development

Research and development (R & D) is a process intended to create new or improved technology that can provide a competitive advantage at the business, industry, or national level. While the rewards can be very high, the process of technological innovation (of which R & D is the first phase) is complex and risky. The majority of R & D projects fail to provide the expected financial results, and the successful projects (25 to 50 percent) must also pay for the projects that are unsuccessful or terminated early by management. In addition, the originator of R & D cannot appropriate all the benefits of its innovations and must share them with customers, the public, and even competitors. For these reasons, a company's R & D efforts must be carefully organized, controlled, evaluated, and managed.

Objectives and Types of R & D

The objective of academic and institutional R & D is to obtain new knowledge, which may or may not be applied to practical uses. In contrast, the objective of industrial R & D is to obtain new knowledge, applicable to the company's business needs, that eventually will result in new or improved products, processes, systems, or services that can increase the company's sales and profits.

The National Science Foundation (NSF) defines three types of R & D: basic research, applied research, and development. Basic research has as its objectives a fuller knowledge or understanding of the subject under study, rather than a practical application thereof. As applied to the industrial sector, basic research is defined as research that advances scientific knowledge but does not have specific commercial objectives, although such investigation may be in the fields of present or potential interest to the company.

Applied research is directed towards gaining knowledge or understanding necessary for determining the means by which a recognized and specific need may be met. In industry, applied research includes investigations directed to the discovery of new knowledge having specific commercial objectives with respect to products, processes, or services. Development is the systematic utilization of the knowledge or understanding gained from research toward the production of useful materials, devices, systems, or methods, including design and development of prototypes and processes.

At this point, it is important to differentiate development from engineering, which can be defined as utilization of state-of-the-art knowledge for the design and production of marketable goods and services. In other words, research creates knowledge and development designs, and builds prototypes and proves their feasibility. Engineering then converts these prototypes into products or services that can be offered to the marketplace or into processes that can be used to produce commercial products and services.

R & D and Technology Acquisition

In many cases, technology required for industrial purposes is available in the marketplace, usually for a price. Before embarking on the lengthy and risky process of performing its own R & D, a company should perform a "make or buy" analysis and decide whether or not the new R & D project is strategically and economically justified. The following influencing factors should be considered: proprietariness, timing, risk, and cost.

PROPRIETARINESS. If a technology can be safeguarded as proprietary—and protected by patents, trade secrets, nondisclosure agreements, etc.—the technology becomes exclusive property of the company and its value is much higher. In fact, a valid patent grants a company a temporary monopoly for 17 years to use the technology as it sees fit, usually to maximize sales and profits. In this case, a high-level of R & D effort is justified for a relatively long period (up to 10 years) with an acceptable risk of failure.

On the contrary, if the technology cannot be protected, as is the case with certain software programs, expensive in-house R & D is not justified since the software may be copied by a competitor or "stolen" by a disloyal employee. In this case, the secret of commercial success is staying ahead of competition by developing continuously improved software packages, supported by a strong marketing effort.

TIMING. If the market growth rate is slow or moderate, in-house or contracted R & D may be the best means to obtain the technology. On the other hand, if the market is growing very fast and competitors are rushing in, the "window of opportunity" may close before the technology has been developed by the new entrant. In this case, it is better to acquire the technology and related know-how, in order to enter the market before it is too late.

RISK. Inherently, technology development is always riskier than technology acquisition because the technical success of R & D cannot be guaranteed. There is always the risk that the planned performance specifications will not be met, that the time to project completion will be stretched out, and that the R & D and manufacturing costs will be higher than forecasted. On the other hand, acquiring technology entails a much lower risk, since the product, process, or service can be seen and tested before the contract is signed.

Regardless of whether the technology is acquired or developed, there is always the risk that it will soon become obsolete and be displaced by a superior technology. This risk cannot be entirely removed, but it can be considerably reduced by careful technology forecasting and planning. If market growth is slow, and no winner has emerged among the various competing technologies, it may be wiser to monitor these technologies through "technology gatekeepers" and be ready to jump in as the winner emerges.

COST. For a successful product line with relatively long life, acquisition of technology is more costly, but less risky, than technology development. Normally, royalties are paid in the form of a relatively low initial payment as "earnest money," and as periodic payments tied to sales. These payments continue throughout the period of validity of the license agreement. Since these royalties may amount to 2 to 5 percent of sales, this creates an undue burden of continuing higher cost to the licensee, everything else being equal.

On the other hand, R & D requires a high front-end investment and therefore a longer period of negative cash flow. There are also intangible costs involved in acquiring technology—the license agreements may have restrictive geographic or application clauses, and other businesses may have access to the same technology and compete with lower prices or stronger marketing. Finally, the licensee is dependent upon the licensor for technological advances, or even for keeping up to date, and this may be dangerous.

Moving Ahead With R & D

Once the decision has been made to perform R & D, the company should decide where and how such R & D should be carried out. There are various possibilities: in-house R & D in the company laboratories, externally contracted R & D, and joint R & D. In-house R & D commands a strategic advantage, since the company is the sole owner of the technology and can protect it from unauthorized uses. In addition, since R & D is basically a learning process, the company can develop a group of experienced scientists and engineers that can be employed in developing more advanced products and processes and in transferring the results of their R & D to operations and to customers. However, since R & D personnel do not like to work alone and are stimulated by peers, the laboratory should have a critical mass in the core technologies and support services; this critical mass may exceed the company resources.

External R & D is usually contracted out to specialized nonprofit research institutions or to universities. The advantages are that these institutions may already have experienced personnel in the disciplines to be researched, as well as the necessary laboratory and test equipment. This will save money and especially time with respect to in-house R & D. The disadvantages are that the company will not benefit from the learning experience, and may become overly dependent on the contractor. Also, the technology transfer may be difficult, and there is always the possibility of leaks to competitors. In the case of universities, costs are usually lower, and there is the additional benefit of identifying graduate students who may be hired later and researchers who may be employed as consultants when needed.

Joint R & D became popular in the United States after antitrust laws were relaxed and tax incentives were offered to R & D consortia. In a consortium, several companies with congruent interests join together to perform R & D, either in a separate organization or in a university. The advantages are lower costs, since each company does not have to invest in similar equipment; a critical mass of researchers; and inter-change of information among the sponsors. The disadvantages are that all the sponsors have access to the same R & D results. However, because of antitrust considerations, the R & D performed must be precompetitive, and each participant in the joint R & D must apply separately the information obtained to its products, processes, and services.

R & D Project Selection, Management, and Termination

Industrial R & D is generally performed according to projects (i.e., separate work activities) with specific technical and business goals, assigned personnel, and time and money budgets. These projects can either originate "top down" (for instance, from a management decision to develop a new product) or "bottom up" (from an idea originated by an individual researcher). The size of a project may vary from a part-time effort of one researcher for a few months with a budget of thousands of dollars, to major five- or ten-year projects with large, multidisciplinary teams of researchers and budgets of millions of dollars. Therefore, project selection and evaluation is one of the more critical and difficult subjects of R & D management. Of equal importance, although less emphasized in practice, is the subject of project termination, particularly in the case of unsuccessful or marginal projects.

SELECTION OF R & D PROJECTS. Normally, a company or a laboratory will have requests for a higher number of projects than can be effectively implemented. Therefore, R & D managers are faced with the problem of allocatingscarce resources of personnel, equipment, laboratory space, and funds to a broad spectrum of competing projects. Since the decision to start on an R & D project is both a technical and a business decision, R & D managers should select projects on the basis of the following objectives, in order of importance:

1. Maximize the long-term return on investment;
2. Make optimum use of the available human and physical resources;
3. Maintain a balanced R & D portfolio and control risk;
4. Foster a favorable climate for creativity and innovation.

Project selection is usually done once a year, by listing all ongoing projects and the proposals for new projects, evaluating and comparing all these projects according to quantitative and qualitative criteria, and prioritizing the projects in "totem pole" order. The funds requested by all the projects are compared with the laboratory budget for the following year and the project list is cut off at the budgeted amount. Projects above the line are funded, those below the line delayed to the following year or tabled indefinitely. Some experienced R & D managers do not allocate all the budgeted funds, but keep a small percentage on reserve to take care of new projects that may be proposed during the year, after the laboratory official budget has been approved.

EVALUATION OF R & D PROJECTS. Since R & D projects are subject to the risk of failure, the expected value of a project can be evaluated according to the following statistical formula:

EV=P×pt×pc×pf

where P is the payoff if the project is successful; that is, the stream of net income accruing to the company over the life of the new product (or process, or service) resulting from the project. The payoff P is then multiplied by the probability of success, which is the product of three separate probabilities:

1. pt is the probability of technical success, i.e., that the new product or process will meet the technical and functional specifications
2. pc is the probability of commercial success, i.e., that the new product will be accepted by the marketplace and will achieve the forecasted market share
3. pf is the probability of financial success, i.e., that the new product will achieve the forecasted financial goals, in terms of profits, return on investment, and cash flow.

Consequently, project evaluation must be performed along two separate dimensions: technical evaluation, to establish the probability of technical success; and business evaluation, to establish the payoff and the probabilities of commercial and financial success. Once the expected value of a project has been determined, it should be divided by the forecasted cost C of the project, in order to obtain a benefit/cost ratio R of the form R EV/C. Obviously the higher this ratio, the more desirable the project.

For more advanced and longer term projects, leading to major (rather than incremental) innovation, it may be difficult to establish reliable values of P, C, pt, pc, and pf. In this case, a relative comparison of projects is made based on their respective technical quality and potential business value. Technical quality is evaluated by analyzing and rating the clarity of the project goals; the extent of the technical, institutional, and market penetrationobstacles that must be over-come; the adequacy of the skills and facilities available in the laboratory for carrying out the work; and how easily can the project results be transferred to an operation. Potential business value of a project is defined in terms of the market share of an existing market that can be captured by the new product; or by the size of a new market that can be developed by the new product; or by the value of new technology that can be sold by the company or transferred to its customers.

After the first tentative list of projects has been established in order of priority, it is "matched" with the existing laboratory human and physical resources to make sure that these resources are well utilized. In fact, creative human resources are the laboratory's most valuable asset, and these should not be wasted by asking researchers to do work outside their disciplines and interests. Also, it is difficult to change in a short time the "mix" of available disciplines and equipment, and to hire and fire researchers. Thus, a shift towards new disciplines should be done gradually, avoiding the underutilization or overloading of the existing resources.

Once the tentative list of prospects has been modified according to the above, the entire project portfolio of the R & D laboratory should be balanced, in order to control risk, according to three types of probabilities listed above. Technical risk is controlled in two ways:1) by having a spectrum of projects ranging from low to medium-high technical risk; 2) by avoiding "bunching" too many projects in the same technology, particularly if the technology could be replaced by a superior technology during the expected lifetime of the new product.

Commercial risk can be controlled by not having "too many eggs in one basket," that is, by targeting different market segments (government, capital equipment, consumer, industrial, international, etc.) and attacking different competitors, since directly targeting a major competitor may trigger a dangerous counter offensive and a price war. Financial risk is controlled by having a majority of small- and medium-size projects (in terms of R & D expense), a few large projects, and no projects that, in case of failure, could bankrupt the company. Financial risk, in terms of cash flow, is also controlled by having a spectrum (in time-to-payoff) of more short- and medium-term projects than long-term projects. This type of spectrum is also psychologically important to maintain the credibility of the R & D laboratory in the face of upper-level executives who keep asking "What have you done for me lately?"

By definition, R & D is a risky activity, and there are no "zero risk" R & D projects, since these would then be engineering projects. While the majority of the projects in an R & D portfolio should be categorized as low-risk, some medium-risk projects are justified, and even a few high-risk projects, provided their expected value is high.

Finally, project evaluation and selection should be made objectively, in order to develop and maintain a favorable climate for creativity and innovation. Researchers will be naturally disappointed when their projects are not approved. Some may even suspect that other projects were preferred for subjective reasons, such as the "halo" effect (the past track record and prestige of other, more senior, researchers), the reluctance of management to terminate less deserving projects, and especially political influences to select "pet" projects of executives. If there is a feeling that project selection is not done objectively, many researchers, particularly the junior ones, will lose their enthusiasm and renounce proposing new projects of a high potential value for the company. Eventually, if this situation persists, the laboratory will lose its creativity and concentrate on routine low-risk (but also low-payoff) or "political" projects, and it will have difficulty in keeping and attracting the best researchers. Therefore, it is desirable that the project evaluation and selection criteria be properly explained and that all researchers be asked to participate in the evaluation process. Also, the finalized project portfolio should be presented to, and discussed with, all the researchers.

MANAGEMENT OF R & D PROJECTS. The management of R & D projects follows basically the principles and methods of project management. There is, however, one significant caveat in relation to normal engineering projects: R & D projects are risky, and it is difficult to develop an accurate budget, in terms of technical milestones, costs, and time to completion of the various tasks. Therefore, R & D budgets should be considered initially as tentative, and should be gradually refined as more information becomes available as a result of preliminary work and the learning process. Historically, many R & D projects have exceeded, sometimes with disastrous consequences, the forecasted and budgeted times to completion and funds to be expended. In the case of R & D, measuring technical progress and completion of milestones is generally more important than measuring expenditures over time.

TERMINATION OF R & D PROJECTS. Termination of projects is a difficult subject because of the political repercussions on the laboratory. Theoretically, a project should be discontinued for one of the following three reasons:

1. There is a change in the environment—for instance, new government regulations, new competitive offerings, or price declines—that make the new product less attractive to the company;
2. Unforseen technical obstacles are encountered and the laboratory does not have the resources to overcome them; or
3. The project falls hopelessly behind schedule and corrective actions are not forthcoming.

Due to organizational inertia, and the fear of antagonizing senior researchers or executives with pet projects, there is often the tendency to let a project continue, hoping for a miraculous breakthrough that seldom happens.

In theory, an optimal number of projects should be initiated and this number should be gradually reduced over time to make room for more deserving projects. Also, the monthly cost of a project is much lower in the early stages than in the later stages, when more personnel and equipment have been committed. Thus, from a financial risk management viewpoint, it is better to waste money on several promising young projects than on a few maturing "dogs" with low payoff and high expense. In practice, in many laboratories it is difficult to start a new project because all the resources have already been committed and just as difficult to terminate a project, for the reasons given above. Thus, an able and astute R&D manager should continuously evaluate his/her project portfolio in relation to changes in company strategy, should continuously and objectively monitor the progress of each R&D project, and should not hesitate to terminate projects that have lost their value to the company in terms of payoff and probability of success

 Reference: Gale Encyclopedia of Small Business: Research and Developmen

Deviation Controls in Pharmaceutical R&D

 

Background

There are numerous examples of products that took many years to research, develop, get approved, and finally, marketed, due in part to inadequate controls during the R&D phases. Products have been approved but could not be marketed because they could not be manufactured reproducibly according to the conditions of approval. Review of data from preclinical, clinical, and other product studies and investigations into discrepant results from laboratory and clinical studies have revealed wide variations in product formulation, handling, manufacturing, and testing.

In some cases, there have been so many issues to consider that causation of variability in study results could not be determined. In other cases, the lack of adequate documentation prevented any meaningful review or investigation because an accurate history of R&D could not be reconstructed.

Too often, it is virtually impossible to accurately trace the progression of a product’s development from initial phases of research, through multiple development phases, and finally, to the product submitted for approval and proposed for marketing. Sadly, since the review of product development typically occurs long after the work is completed, problems are identified late in the product life cycle and can result in redoing multiple studies.

Product Life Cycles

Various authors have written about product life cycles, which include, generally, the following phases: concept, planning, research and development, scale-up, launch, and maintenance. Applying the definition presented above, research can occur during any of the phases of a product’s life cycle although, admittedly, the R&D activities can vary in scope and extent depending upon purpose.

Some R&D activities may be regulated directly by FDA according to GxPs, while others may support further work that is submitted to and ultimately reviewed and evaluated by the agency. Research scientists may assist in the investigation of manufacturing or laboratory related failures as well as the design and development of corrective actions for marketed products just as they design and develop new products and associated specifications, manufacturing processes, and controls.

Various best practices apply to research, development, manufacture, and control of products regulated by FDA and include GLPs, GCPs, GTPs, GMPs, and Part 11 (electronic records). FDA regulations and guidelines related to GxPs include, among other things, requirements for standard operating procedures. Deviations from such procedures must be identified, investigated, and corrective actions implemented to prevent recurrence. FDA-regulated companies generally understand that these requirements exist even if they do not always meet current standards in their execution, as evidenced by the content of numerous Warning Letters issued during the past several years.

Simply, a deviation is a difference from what is expected or planned. In the GMP world, as the manufacturing environment is sometimes called, the concept of a deviation is pretty straightforward. Since the GMP world is highly regimented and monitored, expectations are (or should be) clearly stated in procedures and specifications. (For example: Mix for 10 minutes.) Deviations from these expectations are, therefore, relatively easy to identify.

R&D Environment

But what about the R&D environment where experiments and investigations are conducted to obtain additional knowledge or confirm previous results? Clearly, and appropriately in these circumstances, the results cannot or should not be completely predictable. Nonetheless, such work does contain some expectations. For example, if an unknown sample is tested against a blank, a standard, or a control, there are expectations about the characteristics of the blank, standard, or control that will be used to evaluate the results from the unknown sample. As research progresses toward product development, and the body of information about a product builds, the list of expectations grows.

Deviation control in such a setting involves controlling the variables that can be controlled. When sources of variability cannot be identified or controlled, the work history (who, what, when, where, how) should be documented sufficiently to stand as institutional memory and a source of intelligence for future planning and study.

In early conceptual work, it may be appropriate to apply basic laboratory controls, as described below, and fully document the conditions under which work was performed. This documentation is invaluable to a company and other scientists as a product moves from the laboratory into toxicology studies and clinical trials, and provides a sound base from which to evaluate the impact of both subtle and distinct differences in a product through the development phases.

The validity of conclusions made from the interpretation of experimental data and other test results is built upon the level of control of known sources of variability: people, equipment, methodology, and samples. The more these elements are managed to prevent or reduce variability, the more assurance there is that the data accurately reflects the true characteristics of the product.

Recommendations

Basic controls for R&D

Establish a risk-based quality system for R&D work that is designed to control what needs to be controlled, rather than to set controls just for the sake of setting controls.

Establish a quality system in which the levels of control increase as a product progresses through the development phases.

Institute appropriate documentation practices that provide flexibility for scientists but result in a complete and accurate history of purpose and scope of work; how the work was performed; where the work was performed and by whom; and observations, results, or conclusions.

Establish mechanisms whereby related pieces of work can be linked or cross-referenced so that the story of product development can be assembled efficiently.

Basic controls for projects.

Develop and follow an experimental plan (part of a project plan) that sets forth the specific types of controls deemed appropriate according to the purpose and scope of the work that is planned.

Establish appropriate controls over known sources of variability including:

People Establish procedures describing how work should be performed. These procedures may be presented in memoranda, protocols, SOPs, or other forms of documentation. Provide training, instructions, or demonstrations for employees to ensure that procedures are interpreted and will be followed correctly and consistently. It is particularly important to ensure that technique-based skills (e.g., pipetting) are consistently applied. Do not assume that individuals with equivalent years of education and experience will perform critical functions or interpret instructions in the same manner.

Equipment Use clean or new equipment, accessories, and other lab ware; use instruments that are in good working order and calibrated; specify as many equipment operating conditions as possible and document the actual conditions used or observed.

Methodology Describe analytical methods and manufacturing instructions as thoroughly and specifically as possible.

Samples and other materials Describe and characterize samples, controls, and ingredients in manufactured products according to the scope and purpose of the work. At a minimum there should be a record describing: source, appearance, storage, and handling conditions. As development progresses, and specifications are developed, further testing and characterization is expected.

Applying these simple principles and recommendations will improve the quality of R&D work, add credibility and validity to study results, and help to streamline the product development process.