Thank you, ladies and gentlemen. Please be seated. All right. Let the record reflect that we have been rejoined by all the members of our jury g morning, ladies and gentlemen.

THE JURY: Good morning.

All right. The Defense may call their next witness.

Your Honor, the Defense calls Dr. Fredric Rieders.

All right.

Please raise your right hand. You do solemnly swear that the testimony you may give in the cause now pending before this court, shall be the truth, the whole truth and nothing but the truth, so help you God.

I do.

Please have a seat on the witness stand and state and spell your first and last names for the record.

Good morning, your Honor.

Good morning, doctor. Could you state and spell your name, sir.

My name is Fredric, F-R-E-D-R-I-C, Rieders, R-I-E-D-E-R-S.

Mr. Blasier.

Thank you, your Honor. Good morning, ladies and gentlemen.

THE JURY: Good morning.

DIRECT EXAMINATION BY MR. BLASIER

Dr. Rieders, when and where were you born?

I was born in 1922 in Vienna, Austria.

What is your occupation?

I'm a forensic toxicologist and director of a toxicology laboratory.

Could you tell us what toxicology is?

Toxicology is the study of chemical substances and of presumably biological systems in terms of focusing on the harmful effects that the chemicals can produce in the biological system. Simplified version, toxicology is the study of poisoning, and I deliberately say poisoning in quotes, rather than poison, because everything is a poison in too large a quantity. The definition of a poison is that it is too much.

Doctor, could you please tell us about your educational background.

Excuse me. Doctor, would you pull the microphone down, please.

There we go. Perfect. Thank you. Mr. Blasier.

I studied--my undergraduate studies were at New York University's Washington Square College, at Columbia University, at the University of Indiana in Bloomington, at Houston University and a little bit on courses while I was in service overseas by correspondence from the University of Wisconsin. I received a bachelor's of arts degree from New York University's Washington Square College in New York in 1948. My major subject was chemistry. My minor subject was biology. I then went on towards--I went to a master of science degree in chemistry in which I specialized in toxicological analysis chemistry, in the analysis that is used at that time, essentially in autopsy specimens. After I received a master's degree and became eligible to be a doctoral candidate at New York University, I was offered a special fellowship at the Jefferson Medical College of Philadelphia, which is part of Thomas Jefferson University in Philadelphia in their school of graduate studies, not towards an MD degree, but towards a doctoral, Ph.D. degree. And there during the following, oh, two years of work and one year of this, that and the other thing, I received the degree of doctor of philosophy, not medicine, Ph.D. degree, my major subject was pharmacology, toxicology; pharmacology being the study of the effects of chemical substances from no effect to through toxicology to a lethal effect, as well as what the body does to the chemicals as it is in toxicology. And my minor subjects were pathology, the study of disease and physiology, the study of the function of organisms, essentially the functions of the human organism of man. That was my formal education. Of course I did research and presented a thesis on the toxicology of a substance called acrylonitrile and I was granted a degree in 1952.

Spell acrylonitrile for the court reporter, please.

Sir?

Can you spell acrylo--

Yes. A-c-r-y-l-o-n-I-t-r-i-l-e.

Thank you.

Could you describe your other academic experience and affiliations.

Yes. While I was a graduate student at Jefferson, I was also a graduate assistant and I did laboratory demonstrations in toxicology for the medical students. I did work for the clinic because I had some techniques from my previous work experience in EDTA determination in blood and other toxicological analyses, which were very rare at the time, not many had them, so assisted in this sort of thing. I also participated in other research besides my own doctoral research. After I received the degree, I was appointed an instructor in the department of pharmacology and in industrial medicine as an assistant instructor and I taught medical students, second year medical students, toxicology, both in the laboratory and at lecture. I worked with graduate--other graduate students and I conducted research in various areas of pharmacology and toxicology. And I lectured, of course, as I mentioned. This went on until 1956, at which time I had been promoted to assistant professor and had done research in pharmacology, toxicology and industrial medicine, and that, incidentally, included a lot of research with EDTA at the time, which had just come into use for the treatment of certain types of metal poisons. In 1956 the city of Philadelphia changed from a coroner system to a medical examiner system and the first medical examiner, Dr. Joseph Spellman, asked me to apply for the position of chief toxicologist upon the recommendation of my former chief, Dr. Alexander Gettler, who is sort of a granddaddy of American toxicologists, whom I had worked for in Belleview. So I became an instructor and then I left the employ of the city. I took an examination with several other people. I scored appropriately and was selected for the position of chief toxicologist for the city of Philadelphia and director of--well, started a poisoning information center and directing it.

Let me back up just a minute before we get into your employment history if we can. Have you told us about all of your teaching experience or academic experience with respect to professorships?

Well, only up until that I remained on the faculty at Jefferson. I still am. I am a full professor now. I became associate professor and a full professor in the course of time on a part-time basis, of course, and when the money crunch came, I agreed to forego my salary, which was minimal, which was a stipend, and I'm still on the faculty pro bono, which means I don't get paid for it. Now for the last few years I have not lectured to medical students, but I have worked with graduate students, continue to work with graduate students, and I am starting a whole new course where we will start lecturing in the fall and in the spring.

Now, before we get into your employment history, could you tell us whether you have any affiliations with professional associations?

Yes. The first professional association which I joined was American Chemical Society of which I am still a full member, as well as several of its divisions, including the division on analytical chemistry. Then later on I joined several other societies that were particularly involved in pharmacology; American Association--American Society of Pharmacology and Experimental Therapeutics. I am a full member of that.

Some local societies, which since I no longer stayed active, I dropped from, but I became a member of the American Academy of Forensic Science. I am now a fellow. In particular of the section on toxicology on forensic toxicology of which I was the--was one of the chairmen for one of the sessions some years ago and had various other official capacities in the academy since about 1958 or `59, since I have been a member there. I am a member of--and I am one of the founders, one of the charter members of the International Association of Forensic Toxicologists which was formed, I think it was around 1960, at a meeting in England. I was the first editor of its bulletin, which is after finding somebody with a printing press made himself available, I ran on a Xerox machine for a couple of years. I am a member of the--well, of several other organizations. I am a member of the American Association of Forensic Toxicologists, which is called society of forensic toxicology, which is somewhat similarly composed and somewhat similar to the activities of the toxicology section of the academy of forensic science. And I am--I got certified by the American board in forensic toxicology, which is part of the academy, as a fellow, as a board certified forensic toxicologist. I am also a member of the association--American Association of Clinical Chemists. Those are the chemists who work in hospitals and in other places mainly to help in the diagnosis of disease. There is a section, an area, and it is a specialty of toxicology, of toxicological chemistry, and I am board certified in toxicological chemistry by the American Association of Clinical Chemists. I am a member of an International Association on Risk Analysis and several others, but these are my main professional background organizations.

Now--now, could you tell us a little bit about--briefly about your employment history prior to the time that you started your own lab.

Well, I won't trouble you with all the different jobs I had--jobs I had when I came here as an immigrant, from stevedore to glass blower to anything you could do in 1939, but my first related position developed in 1941 when I became an assistant chemist, of all things, in a soda factory in preparing flavors and doing preparatory analyses. After that I worked for another chemical company that made detergents and a variety of other things as an assistant chemist and then I went into the service.

I was in the military during world war ii as a surgical technician and then as an interpreter because of my various language skills. That was towards--the interpreter was toward the end of the war, and then for a year or so afterwards in military government. When I returned to the states I continued under the GI Bill with my college education, but after one year all I had left were some rather easy courses, so I sought employment and I received a job through the employment service as a junior toxicologist, a trainee in toxicology in the office of the chief medical examiner of the city of New York and under the chief toxicologist, under Alexander Gettler and Dr. Charles Hamburger. I was there for approximately two, two and a half years being trained there and actually performing service work, of course. I then, as I mentioned, went to Jefferson and I took you through the time that I left my full-time status as--at that time assistant professor and became chief toxicologist for the city of Philadelphia. My job at that point in time was to organize a laboratory in the division of toxicology. I was not in the medical examiner's office because the medical examiner had just been established. And also to organize and direct the poisoning information center for the city. After that, what I did is the medical examiner's office was in the health department of the city. I conducted toxicological analyses on autopsy cases for the medical examiner, assisted hospitals in the differential diagnosis of poisoning. At that time that is about the only source they had for laboratory work is to avail themselves of the medical examiner's toxicologist. I did some consultive work for a variety of city agencies and I started to train and conduct research in toxicology as well. I did that from 1956, when I became chief, and I--and I organized and directed the poisoning information center. I did that until 1970 for fourteen years and--

Is chief--is office of the medical examiner, that is similar to what we call a coroner?

Very similar, yes. I--we--the operation that was here, the toxicologist that was here was a colleague of mine that I knew very well for many years, and the medical examiner--the coroner I knew very well, and I still know all the people quite well in the ME's office here. It is very similar. I think you have a coroner's medical examiner or a coroner's chief or forensic pathologist, so it adds up to the same thing. It is just politically a different issue. The medical examiner is appointed upon examination.

Excuse me, doctor. This is not your employment history. Ask another question, please.

Can you tell us from 1956 to 1970, when you were with the medical examiner's office, approximately how much of your work involved working with law enforcement agencies?

Well, it all related to law enforcement agencies. We had a weekly conference with the chief of the D.A.'s homicide division, with the chief homicide detective in the city of Philadelphia going to all the homicide cases, but the office was independent of the law enforcement agencies, is a very important part of its function, and that is to be an ombudsman in cases for the medical examiner, not the toxicologist, to determine the manner of death, whether or not criminality might be involved.

Now, from 1970 to now what has your employment been?

Well, in 1970 I left the employment of the city and started an independent laboratory and consultative (Sic) company, national medical services, in which at that time I continued to do a lot of postmortem toxicology for the surrounding counties--

I will get into the specifics of that, but how large is national medical services now?

Well, now it has 120 people in it. It is not my doing; that is my son's.

And does that--does that company do business with law enforcement agencies?

Yes, indeed.

Approximately what percentage of the work is done for law enforcement agencies?

The forensic work that we do is perhaps as much as twenty percent of our work.

Could you give me just a real rough list of some of the law enforcement agencies that you work for.

Well, we work for many of the police departments in the surrounding counties, for the District Attorney's office, for cases that they bring to us in the surrounding counties. We occasionally work with the police department and the courts in the city of Philadelphia, too. We are scientists, so we assist there. Well, either the courts themselves or sometimes the Defense attorneys. You know, it makes no difference.

Do you work with--

We also do work for police departments at times in new jersey, in as far away--well, we have done work for the Los Angeles Police Department at one point or we may have done some more recently that I don't know of.

Do you work--

So we do it for all over the country.

Do you do work for any foreign police agencies?

Yes. Occasionally we did some work for the medical examiner and the Prosecutor in Brazil in one of the countries. Puerto Rico of course, which is part of the United States, we have done work there. We have received specimens in three criminal cases, potential criminal cases, from Australia, one of which is specifically from a police department, the other one is from a medical examiner, the third one a private party, recently.

Does your lab do work for other labs?

It is--a major part of our work is reference work. That means work that large laboratories, which do a lot of clinical analysis, get and there are specialized toxicological tests and they send them to us, we are their reference lab.

Can you give us an estimate as to how much of your work is--other labs have sent to you?

I can only do it for the time in which I was administratively involved in the shop, which stopped about three or four years ago, and at that time it was about thirty percent or forty percent was from reference work from other laboratories.

Now, do you have an ongoing working relationship with the FBI?

Yes.

Now, I don't want to mention any specific cases, but are you currently working with agent Roger Martz of the FBI on some criminal cases?

Well, we are looking--not actually working on them, but we are discussing how to work on them that we have, yeah.

Does that work involve detecting or developing methods to detect the presence of poisons in tissue?

In one case it really involved the development of methods. In the other case it will be what analysis will the FBI do or has done already and what analyses will I do because of my capabilities.

And this--special agent Roger Martz is sitting right behind me, right?

Yes, yes, of course.

Now, could you tell us, very briefly, when you started doing work with chromatography?

Well, I started doing work with paper chromatography, which was the first form of chromatography that came out, while I was still a graduate student, so that I was in still--let's say 1950. I started doing thin layer chromatography, which is a more modern method and used it extensively following a Gordon research conference in which I participated in 1959 or `60. Same year started working with gas chromatography; 1959, 1960. I started to work with gas chromatography coupled hyphenated--you know, the hyphenated systems with a technique called mass spectrometry, oh, in the seventies.

Now, was that around the time when that technology actually started?

Well, the technology started earlier as an academic research technology, but it is around the time where they were beginning to be some bioanalytical applications for real life situations.

Now, have you done work in what's called HPLC or high pressure liquid chromatography?

Yes, yes, I have.

Since about what time?

1973--I think 1973 is when we got our first instrument after I had familiarized myself with it for sometime prior thereto and so that is about when I started to actually hands-on do a lot of work on it and then supervise a lot of work.

Do you currently in your lab do mass spectrometry?

Oh, yes, we do.

And how many--how many instruments do you have that do mass spectrometry?

Seven or eight.

Now, we will get into the details of what that is in a second, but have you done what's called tandem mass spectrometry?

Yes, I have.

Approximately how many times?

A few times, hands-on only.

Okay.

Again, that is a relatively new not commonly available technique, in toxicology labs at least.

Could you tell us briefly what is EDTA?

Well, EDTA is the name of a chemical substance which in full is called ethylenediamineacetic, e-T-H-Y-L-E-N-E, diamine, D-I-A-M-I-N-E, acetic, A-C-E-T-I-C, acid. Ethylenediamineacetic acid.

Your Honor, could we put up slide no. 1 in my--

This will be the 1257 series. This will be 1257-A.

Doctor, can you see the monitor right there to your right?

Yes.

Is that what EDTA is, what you just spelled?

Yes.

Okay. Could you tell us what a purple or lavender-topped tube is?

That name is in medicine and in laboratory medicine or anyone who uses human blood for testing, the name for an evacuate tube which has a stopper that is--some call it lavender, some call it purple in color, and that makes you recognize it clearly as a tube which has EDTA in it.

Your Honor, I would like to have a photograph marked as next in order.

1258.

All right.

I would like to have this put on the elmo, please.

This is a photo of tubes?

Yes.

1258, Mrs. Robertson, a photo of tubes.

Doctor, could you look at your monitor and tell us if that appears to be purple-topped tubes?

That is the typical appearance of the purple-topped tubes, yes.

Now, what is the relationship between EDTA and a purple-topped tube?

EDTA, either a potassium or a sodium salt, is put into the purple-topped tubes which have a vacuum in them so that when blood is drawn and mixed with them that that blood won't coagulate. That is the sole purpose. It has some preservative purposes for certain types of cells and other things, but what the EDTA does, the EDTA is what is called in English a claw compound like a lobster's claw, a chelating agent. And what the excess of EDTA does that is in the tube is combine with all of the calcium in the blood and tie it up. Without calcium blood won't clot. If you don't do that, then the blood will clot in five, seven minutes, you know.

Now, you had experience in your background working with EDTA?

Great deal, yes.

And approximately when did do you your first work with EDTA?

In the early fifties.

Just briefly, what was the nature of that work?

The first thing that I became involved in is to study its properties as to whether or not it could be used for treating acute lead poisoning in small children who were at the Jefferson hospital and then in working with one of the physicians in administering and designing doses, administering, collecting blood in urine and measuring the EDTA that is coming out of the child. Then this was expanded to working with it in occupational employment cases where people are exposed to lead, battery workers and others, both in treating this kind of lead poisoning which is different from what is in children, but also we developed a diagnostic test for somebody to determine whether or not someone had an excessive body burn of lead and therefore should be treated even before they got really sick. So all that was published work and then I did a substantial amount of test-tube work with EDTA and also experimental work with animals. I also did experimental work, or rather studies, it wasn't really experimental, it was clinical work, in determining how EDTA mobilizes and causes the excretion of other elements besides lead, again with published work for copper, for zinc, for a number of other metals that EDTA can chelate. In the process--well, there was the biological work. The invitro work was measuring the strength of the way that EDTA holds onto metals, how easily will it rip it away from some part of the body, how it will redistribute the lead and whether it is absorbed, fed in animals; not in man. I did not need--we did not need any people because we came to the conclusion that oral EDTA can only do harm rather than good to people of metal poisoning.

Now, did you have--some of that work involved analyzing biological samples for the presence of EDTA?

Some of it did, yes.

Starting about when?

I made some attempts at analysis of EDTA as early as 1954 or `55. They were successful for qualitative tests in urine but not for anything test. They were color tests of a sort. I did a lot of tests--not a lot--some testing for EDTA while I was toxicologist for the city because every so often I would receive a blood specimen and I needed to know what was the preservative, and the pathologists couldn't help me, didn't remember whether he had dumped this blood from a reagent from a lavender-topped tube or from a gray-topped--a gray-topped tube has fluoride and oxylate in it--or whether he didn't put anything in it. After death blood stays fairly liquid, it clots, but then it liquefies again, so you can't really tell. If you get a tube of clotted blood, you know that is blood that has not been preserved with anything usually. That is why it clots quickly.

Have you worked on chromatography techniques to detect the presence of EDTA?

Yeah. That came later.

That started about when?

Oh, I did--started to do some of that around 1976 or `77, but it isn't for detecting EDTA at ultra trace levels. It is to determine whether a blood specimen is an EDTA specimen, has a lot of EDTA in it, or whether it is a fluoride oxylate specimen, whether it has been preserved with that. More recently I am able to determine whether heparin was used. I couldn't do that until recently.

Doctor, I didn't ask you about your publications before. Have you published articles and made presentations in your area of expertise?

Yes.

Approximately how many?

Over a hundred.

And do you know approximately how many of those--how many of your articles are in peer review journals?

Are what?

How many of your articles appeared in peer review journals?

I would say well over half.

And did some of those articles include articles about EDTA?

Yes.

Approximately how many?

A dozen or so.

Now, were you asked to review results of a study performed by the FBI to determine whether there was EDTA present on bloodstains found on a pair of socks found in Mr. Simpson's bedroom and from a bloodstain found on the back gate of Nicole Brown Simpson's condo?

Yes.

And have you examined test results produced by the FBI on those questions?

Yes.

Have you also examined a validation study that the FBI performed in February of this year?

To the extent to which they were submitted, yes.

Incidentally, at one point were you consulted by the Los Angeles Police Department during the course of this case about methods to detect EDTA in bloodstains?

Yes.

Do you remember who that was?

Mr. Henkhaus, H-E-N-K-H-A-U-S.

And did you provide him with some materials to assist them in developing a method to detect EDTA?

Yes, of course.

Now, do you have an opinion as to whether the method used by the FBI to detect the presence of EDTA in bloodstains is a valid method for detecting the presence of EDTA in blood?

In terms of determining whether it is there or not?

Whether it is there and whether it is there in a range, a very wide range of amounts.

Now, when we talk about parts, do we use the term parts per million?

No, that is concentration.

Okay. Tell us briefly what is--when you use the term "Parts per million," what does that mean?

It means one part, in the case of EDTA, in a million parts of blood. This would mean, for instance, one microgram per milliliter which is one gram or a million micrograms of blood. That is one part per million.

Did the method developed by the FBI that you reviewed, in your opinion is it capable of detecting amounts of EDTA in blood in the parts per million range?

Yeah. In the range of maybe ten parts per million and higher, perhaps somewhat less than that. Perhaps five parts per million and higher.

Now, is parts per billion one thousandth of parts per million?

It is one billion and a billion is a thousand million.

If you are relating parts per million to parts per billion, you are talking about 1000 as much of something?

Right. There are a thousand parts per million in one part--I mean there are a thousand parts per billion in one part per million. They are the same. A thousand parts per billion or one part per million.

Doctor, I want to ask you some questions about EDTA and what characteristics the FBI was looking for. Could we have slide C.

1257-C.

Now, do chemical compounds like EDTA have what is called a molecular weight?

Yes, sure.

And is this--this isn't in pounds or ounces, is it?

It is in units. The molecular weight is how many hydrogen atoms you can cram into a molecule to get the same weight. That is the unit. The weight of hydrogen is nominally one. So something that weighs--has a molecular weight of 200--292, as we have here, and 292 is the molecular weight, is as big, weighs as much as 292 hydrogen atoms.

So what is the molecular weight of EDTA?

292.

And I assume different compounds have different molecular weights?

Yes. There are other compounds with 292 weight, but they are not terribly many.

Is 292 a relatively high molecular weight?

Relatively high, yes. As you go higher, fewer and fewer compounds will have exactly the same weight.

Now, looking at the chart on the screen, the chart indicates adding a proton to 292, gives you a--gives you what?

293.

And--

The proton is the same as a hydrogen atom with a charge on it, positive charge of hydrogen atom.

And what is the--why do you need to have a charge on it?

Well, you need a charge on an atom in order for the principle of mass spectrometry to measure it. The mass spectrometer measures charged atoms, you know, measures the charges. And what it does, it gives you a number which is the ratio of the mass, the molecular weight over the charge. In this case it would be 293 over 1, so that the ion that you then have, which is EDTA with a positive charge hooked on its head, is--has a weight of 293, and the charge--that is the mass, 293. The charge is called z, like zebra, over 1, so it is 293.

So is it fair to say that if you were looking for EDTA in a substance, one of the things would you look for is whether you had something that weighed 293?

Yes.

And what is that called, 293?

That is called the parent ion. It is the molecular--it is the whole molecule plus a charge on it, so it is an ion. The whole molecule doesn't have a charge on it, but it gets a charge on it and it is called a parent ion because from it can come fragments.

Now do we have the next slide, slide D.

Now, doctor, please look at your monitor, and the bottom part of that diagram now indicates that the parent ion is broken into pieces; is that correct?

Yes. That is the function of the mass spectrometer. The first thing it does, it weighs the ions that--it weighs the molecule. The molecule, in order to be weighed, has to have a charge so it adds a proton and then weighs it, so it gets 293, which is the molecule plus one. The molecule, remember, is 293. Then it breaks that molecule into pieces, but not the way that you break a bottle of beer, but the way that a jeweler cuts a diamond. It breaks it along certain planes and not others so that the pieces that result are somewhat predictable from the structure of the compound. If you apply energy to it, it will break up into certain kind of pieces. Now, what it does, it weighs the pieces to see what weight do they have. In this case one of the pieces, one of the daughters, you might say, of that 292 after it was broken up, has a weight of 160 so that you now have--you have two identifying characteristics. You have--there is a substantial amount among all of the ions of the 293, enough so you can weigh it, and there is enough of the break-up product, 160, so you can weigh that.

Now, I want to talk a little bit about chromatograph. Could we have slide F.

Your Honor, the last side was 1257-D.

Thank you.

This is 1257-F.

Now, the process that was used by the FBI in this case involved a step called chromatography and then a step called mass spectrometry, correct?

Yes.

Now, chromatography, could you describe briefly, looking at the chart up there, what is the principle of chromatography?

The principle of chromatography is the separation of compounds that are mixed together by having them go for a walk, so it is characteristic--for instance, you take a group of people, a whole bunch of people, and you say now I want you to walk from here to a mile from here as fast as you can. You all start at the same point. Gradually over that mile it spreads out until the EDTA person is somewhere there and all the others are also spread out. Good chromatography separates a mixture into all of its components, or all of the components that you are interested in at least, and then at the end of the mile there is someone with a camera who takes a picture of the person coming through the end gate and determines how long it took from here to the mile from here, and says that is the retention time. That is how long that person was retained on the road to the goal.

Let me go to the next slide and see if we can illustrate that, which will be G.

Now, in my example I have used a red square as an EDTA compound molecule and two other ones, one green and one yellow. The column that is indicated on the diagram, what is that?

That is actually a glass or a metal column which is filled with a material that the person can lean on, so to speak, if you make it parallel to this, so that there are things along the mileway where you can stop and lean on, if you want to, for a little before you go on. What is driving the material through that column is a liquid. In this case, in the illustrating case of people moving as fast as they can, the drive is more inward, you know, if you want to win or you just want to make it to the other end, but what is moving you is your muscles to gravitate and some people lean longer and others lean less long and so they start spreading out. So the column is packed with a material and it is--the mixture is pushed through the column with a liquid. That is a steady rate. As the mixture goes through the column, it leaps on and releases from the column the different compounds different number of times, so that again by the end of the column they have separated from each other. In this case the EDTA is the slowest that I have here. It is the last one out of the column. Other things before it came out before.

Now, in the diagram, I have a clock at the top and a clock at the bottom. Is that to represent the amount of time it takes the EDTA to get from the start, go through the column and come out the other end?

Yes. That is the stopwatch and what it measures is the retention time.

And is that one of the things that you look at to determine whether you have EDTA in a substance?

Yes. In a good system you can repeat that. You can take another sample of the mixture and throw it on the column and push it through and the retention time is going to be pretty much the same, close to it, within what you allow as experimental error. It won't be the same to ten decimals in time but it should be close enough so that you can say, well, when I see something coming up at that time, I better think of EDTA, and that--you know, that is what EDTA does, so it could be EDTA.

Now--

It is consistent with it.

--the--can you also put known EDTA into the system to see how much time it takes to get through so you have some sort of a benchmark to--

Well, you start out with known EDTA just to determine its retention time, but since from day-to-day and actually from run to run you can see differences, what you like to use is another substance, which you also put in yourself at the same time, which also has a known retention time, but more importantly, is closely enough related to the EDTA so that if there is going to be a change in retention time for the EDTA, there will be a similar shift for your--for your known substance that is called an internal standard.

So now instead of saying it took two minutes for the EDTA to come out, you say it took twice as long as it does for the--for the control substance, for the internal standard, which took one minute. The next time maybe you really have a very different column and it takes three minutes for the EDTA to come out, it will take one and a half minutes for the internal standard, so the ratio will remain the same. It is a very important part of reproducing your value as closely as you can.

Okay. We will talk more about that in a few minutes, but now the technique that was used by the FBI is called liquid chromatography, correct?

Yeah.

And does that mean that it is a liquid that they push through the column that might have had EDTA in it?

Yes.

The next stage is the mass spectrometry stage, correct?

Well, actually the next stage is getting it from the end of the column to the mass spectrometer.

To be put through the mass spectrometer does it have to be a liquid or a gas?

Well, it has to be either particles that are so fine that they are virtually no bigger than gas molecules or that they can move or it has to be a gas.

And now using liquid chromatography, if it comes out at a liquid, how do they get it to a gas or to a small misting particle?

Well, originally what I--originally what you did is you watched the liquid chromatograph and the detector when the peak started to come out--you know, when one of the molecules or of course a lot of it, because you put in not just one molecule of each but whatever came out, you caught it in a net in a fraction collector and then you took that fraction and then you put it into a tiny glass vial and evaporated off the liquid part of it so that you have a solid in it. Then what you did is you shoved that vial into the mass spectrometer and you heated it up until the material in the little vial on the probe evaporated, formed a gas, and then it could react in the mass spectrometer.

What technique did the FBI us?

The FBI used a method where the liquid goes into a very tiny, tiny, but moderately long piece of glass capillary. It has a tiny--it is a glass tube with a tiny, tiny opening and a tiny run. That is heated and has applied to it a very high, very high voltage and a moderate amount percentage, a high electricity. That is why it is called electrospray. At the same time a gas passes over it, a sheeting gas and a separating gas, and what happens is that the liquid at the end of the capillary is sprayed out and in minuscule form and dry form, the water is removed by the sheeting gas, goes into the mass spectrometer, so it is one way and it is the most recent ways and one of the most efficient ways of getting the sample in there. Before that you turned it into a gas in a chamber and then you put the gas out through a small opening which was partly opened to the open air, so that much of it--much of the carrier gas would go out into the area and the stream of vapor of the molecules that you wanted would go in and that is a much less efficient way of transferring from the column to the mass spectrometer.

As the electrospray process, does it change any of the ions?

No, it doesn't. If it changed the ions, then you basically wouldn't know where you stand.

Now, electrospray, do you use that technique in your lab?

No.

And is that a fairly expensive piece of equipment to use?

Well, yes, of course all equipment is expensive. All of this type of equipment is expensive, but it is of relatively recent origin. It is the host up-to-date transfer of molecules in the Hewlett Packard instrument. And as a commercial laboratory we have to wait until something has proven itself before we put out three quarters of a million dollars, so we are not the first ones but also not the last ones to get in and in time we will now,

I would like to move to slide I and get into the mass spectrometry part of this process.

Now, does the mass spectrometer have a way of looking to see whether the 293 part ion is there?

Yes.

And how does it--just very briefly how does it do that?

Well, basically what the mass spectrometer is, is a--is a piece of equipment, which to characterize it, weighs the particles that are in there, sorts them out into groups, into 293's and others, and then measures--weighs how many of those particles are there, what is the intensity of that ion. Then that is a regular mass spectrometer, single-stage mass spectrometer.

Now, in the kind of testing that was done here is a filtering system set up so that only 293 part ions get through?

Well, then in this case, what you do is you set up a filter and you set it so that only the 293 will get through. That is done both in terms of the mass, but also in terms of charge, and such a filter works quite nicely, so you isolate from the others the 293 ion. Now, what you can do is put it on a detector and say it is present period or you can put it into a second mass spectrometer, but in fairly pure form. You just have 293 moleculars. All the other stuff that is in there is not going with you.

Okay. Let me put up slide J.

Now, is that an indication of--just in chart form, of the parent ion getting through the filter and other ions not getting through?

Right. That is what a filter does. It keeps the other particles out and the 293's go into the next place which is either a detector which says I'm seeing things and the filter is taking out everything except the 293, so what I'm saying is 293.

Now, could we have chart K.

Now, is there a second step in order to look for the daughter ion?

Yeah.

Okay.

Instead of going to a detector only it goes--it is detected but it also is then put into another mass spectrometer by the system, then that mass spectrometer again breaks the 293 the way a jeweler breaks a piece of a diamond. It starts out with a ten karat, he breaks it down to three, two, et cetera, then takes one of the three karats and cuts that further.

Could we have slide l, please.

Now, does this chart indicate what happens when it passes through the second filter, it is broken and then the 160 daughter ion is measured?

Right. Again, there is a filter which focuses, it is a focusing mechanism which filters out other things and pushes them aside and looks for 160 masses. Those 160 masses I gathered together and one by one they hit a detector, so the detector says I see things, I see things, I see things, and they are 160 because the others have been filtered out.

Now, could we go to slide m, please.

So if you were trying to determine whether EDTA was present in a substance, what are the three things that you would look for with this kind of testing?

The first thing that you look for is if I extract something with water and put it through my column that I am using in this experiment the way I have tested EDTA and found that it comes out in five, six minutes or whatever, will I see--will something come out at six minutes? That is the retention time, or whatever the retention time is. Then what I will do is to have a mass spectrometer look at what comes out at that retention time and tell me whether I will focus on the 293, whether there is any 293, which is also a characteristic of EDTA, so the retention time is one characteristic. The 293 that I am looking at, that means I am only looking for orange-eyed people you might say. If I see orange-eyed people, I count them; others I don't count. I take all the orange-eyed people after I have counted them and throw them in the next mass spectrometer and it breaks them up and only leaves the orange eyes which eight--weigh 160 let's say.

So we are looking for those three things?

Well, retention time--no. We look for more than that. We look for water solubility because that is how we got it. We look for it going through the column at all because a lot of things don't go through that column. And also that if it does go through the column, that it takes it as much time and no more and no less than the window that I know will contain EDTA, so that is really in a sense the third parameter. Then comes the--does it contain enough molecular ions, 293 masses, you know, are they enough guys in there with--or gals--with 293 on their shirts so that I can actually count them, because if it is less than a certain number I can't really count them so can I see them? Are they there in any other detection--we know my detection limit? Then after breaking those up, the third parameter is the pieces that have 160 tattooed on them and that is what we are looking for, so that we have essentially five parameters.

Did you want to take a break?

Five minutes.

Okay.

Now, also one of the other things that is measured in this system is what's called ion count?

Yes.

And that is--is that essentially counting the number of ions that are getting through to the 160 stage there?

Well, it is counting the number of ions that the detector can see and see--where it can separate the counts, one from the ion, where he can really count, one, two, three, four, and that means that the detector has to snap at an ion and record it before the next ion gets there, otherwise it will record two as one, so it depends open how quickly it is scanning what is coming in. And if it scans a thousand times a second, it is not going to miss many particles, many 293's, but if it scans it 50 times a second, then it will count 10 as one, you know, like a bunch. So--but that is what it does. It gives you a measure of the amount of that particular 293 or 160 that has come into the mass spectrometer, has been ionized in the first one seen and counted as 293 ions, and then the second one, how many did the detector--how many counts did the detector make.

Now, can we go to the next slide, please, slide N.

In the testing done by the FBI, did you review material indicating that they had tested a swatch from the back gate representing stain no. 117?

Yes.

And what number did the FBI assign to that stain that swatch?

I would have to see the numbering system. I think I have it here. May I refresh my memory?

Sure.

(Witness complies.) Okay. From the back gate? Umm, they gave--they gave that the number Q204, I believe.

Now, the sock stain, is it your understanding that the sock stain that was examined was a cutting from the edge of a large stain on the sock?

Yes.

And what number was that given?

Was that given number Q206?

Yes, Q206.

Now, could we switch to slide p, please.

Well, is it your understanding that--well, there we go--on slide p, what is your understanding from the paperwork prepared by the FBI as to where Q206 came from on the sock?

Q206 was cut from the edge of a stain.

Could you look at the monitor. Does that appear to be a chart showing the approximate location of that cutting?

It is similar to what I had on a piece of paper, yes. It is a good reputation.

And the green area is represented--that is a large cut-out on the sock, correct?

Yes.

That was presumably already there and Q206 was a cutting taken from the edge--

Yes.

--of that large a cutting? Was there also a swatch submitted that had been taken from the large swatch that had been cut that was called Q207?

You mean that was taken from the swatch that didn't have the stain on it?

No, that was taken presumably from the large cut-out piece and sent to the FBI along with the sock or are you aware of that?

The 207?

Yes.

I don't think--I don't think so. I don't see it in my papers here, the Q207. It may be. I would have to look through the pile. You know, it is a big pile and it wasn't submitted in a good order, so I would have to look through the whole thing, but I don't recall it.

Let me ask you this: Did you see any testing done using the method that we have described looking for the 293 parent ion, the 160 daughter ion on any sock stain other than Q206?

No.

Okay. All right. Ladies and gentlemen, we are going to take our mid-morning recess at this time. Please remember all my admonitions to you. We will stand in recess for about 15. Dr. Rieders, you can step down. Come back in fifteen minutes, please.

Thank you, your Honor.

All right.