XA0054839

MINOR AND TRACE ELEMENTS

G.V. lyengar and L Tandon

3 1/25 FOREWORD

As part of a Co-ordinated Research Project (CRP) on Comparative International Studies of Osteoporosis Using Isotope Techniques, the International Atomic Energy Agency (IAEA) has recently supported some studies of trace elements in human . In connection with this work, the need became apparent for an up-to-date literature review of minor and trace elements in human bones. The authors of this report were commissioned with the task of doing this, and were also requested to extend their review to cover human teeth. Their report is reproduced in this form mainly to make it available to the participants in the abovementioned CRP. However, it is hoped that the report will also be of interest to a wider audience. The IAEA would like to express its thanks to the authors. Any comments on the report would be welcome; they should be sent directly to the authors* with a copy to the IAEA's Section of Nutritional and Health-Related Environmental Studies.

Addresses for correspondence:

G.V. Iyengar Biomineral Sciences International Inc. 6202 Maiden Lane Bethesda,MD 20817 USA

L.Tandon 505 Oppenheimer Drive #503 Los Alamos, NM 87544 USA

Section of Nutritional and Health-Related Environmental Studies International Atomic Energy Agency P.O. Box 100 A-1400 Vienna AUSTRIA TABLE OF CONTENTS

1. Introduction 1

2. Basis for data collection 1

3. 2 3.1 Bones in the human body 2 3.2 Characteristics of bone 2 3.3 Methods for sampling bone 4 3.4 Methods for processing bone 4

4. Tooth 5 4.1 The human teeth 5 4.2 Characteristics of tooth 5 4.3 Methods for sampling tooth 5 4.4 Methods for processing tooth 6

5. Analysis 6 5.1 Analytical methods 6 5.2 Reference materials for bone and teeth 7 5.3 Preparation of bone and teeth for measurement 7

6. Elemental composition of bones 8 6.1 Major and minor elements 8 6.2 Trace elements 9

7. Elemental composition of teeth 12 7.1 Major and minor elements 13 7.2 Trace elements 13

8. Discussion 16 8.1 Limitations of the data compiled 16 8.2 Major and minor elements in bones and teeth 16 8.3 Trace elements in bones and teeth 16

9. Concluding remarks 18

10. References 19

ANNEXES (Data compilation)

Table 1: Range of Mean Values for Major, Minor and Trace Elements in Human Bones 23 Table 2: Range of Mean Values for Major, Minor and Trace Elements in Human Teeth 25 Table 3: Major, Minor and Trace Elements in Human Bones 31 Table 4: Major, Minor and Trace Elements in Human Teeth 69 Figures 88 References (for compiled data) 93 1. INTRODUCTION

Bone is a major compartment of the skeletal system. It is a living, dynamic structure which provides a supporting and protective framework for the body. Bone provides a reservoir of calcium and phosphate and also has functions in magnesium metabolism. The core contains marrow, which serves as a reservoir of nutrients and produces several types of blood cells. Bone consists of 35 % mineral salts (chiefly calcium and phosphorus), 20 % organic matrix (of which 95 % is ), and 45 % water. About 99 % of body calcium is found in bone. Besides numerous organic constituents, bone is also a major pool for some trace elements. Trace elements such as F, Sr, Pb and U are predominantly found in bone, and are generally termed as bone seekers.

Bone is an example of a biological sample that presents numerous difficulties in obtaining a specimen for chemical analysis. This problem is acute when investigations are to be conducted to determine total skeletal content of a given analyte. First of all, the basic process of sampling bone can be a very difficult task. With humans, this problem is compounded due to medico-legal implications. Not withstanding these obstacles, the question of which particular bone qualifies to be a representative sample of the as a whole (if at all) is a debatable point. Even if some assumptions are made to answer this question, the sub- compartments of a bone sample, namely cortical, trabecular and the marrow and their relative proportion and significance in relation to bone as an , present intricate situations in making definitive decisions. Choice of multiple types of bones is of course the best solution but it is beset with logistical aspects of procuring many bone specimens depending upon the scope of the investigation.

Therefore, it is not surprising that reliable chemical composition data, particularly for minor and trace elements, are scarce. This is also substantiated by the fact that there are only a couple of certified bone reference materials available for validating analytical methods. However, a survey of the scientific literature shows that some analytical work has been expended to establish the elemental composition profile of bone. The usefulness of such analytical information depends upon whether or not analytical quality control was exercised at least to some degree so that an evaluation of the results can be undertaken to identify reference values.

Arguments, similar to that of bone can also be extended to human teeth, with the exception that some of the medico-legal implications are less restrictive than those surrounding human bone sampling.

The purpose of this report is to screen literature information on the elemental composition data for human bone and teeth and to identify reference range of values where data permit such conclusions. This report will also include a concise assessment of the sampling practices in use, and measurement techniques that are applicable for elemental analysis of bones and teeth.

2. BASIS FOR DATA COLLECTION

There have been a few sources, [1-4] which have attempted to shed some light on the activities taking place in understanding the elemental composition profile of bones. Of these, the publication of The Reference Man by the International Commission on Radiological Protection (ICRP) [1] marks an important milestone in the elemental composition of the human body. This excellent source documents analytical results generated during the years 1950-1970 (mainly) for human tissues including bone, carried out in the mainly in the 60s. However, limitations arising from inadequacies of methodologies practiced at the time ICRP-23 was published were recognized by the analytical community and efforts continued to generate accurate data to improve existing information as well as to cover fresh grounds for those elements for which data were not available. An account of part of that analytical improvements has been summarized in a compilation [5] by expanding the scope of elemental coverage. Since then, further improvements have taken place in analytical approaches, especially with the availability of certified reference materials (CRM) for analytical quality control and therefore, generally speaking the status of trace element analysis in tissue samples is at a stage where reasonably reliable results are being generated. Moreover, many investigations are designed to reflect interdisciplinary perspectives [6] and this has further enhanced the overall quality and validity of the sampled specimen. Finally, sustained application of analytical techniques such as the Inductively Coupled Plasma Mass Spectrometry (ICP-MS) has spurred the field of biological trace element research. Hence a fresh evaluation of the literature results is useful to scrutinize the situation and explore the possibility of evaluating reference values.

3. BONE

3.1 Bones in the human body

The human skeletal system and associated major groups of bones are shown in Figure 1. Temporal, vertebra, ribs, sternum, humerus, ilium, ulna, femur and tibia are examples of sources used to obtain autopsy or biopsy samples. Tibia is a readily accessible bone. Similarly, rib samples are sought at autopsy. Iliac crest is usually the sampling site of choice for histologic examination of trabecular bone.

3.2 Characteristics of bone

Bone is composed of osseous tissue, a tissue made hard by the deposition of inorganic substances in a process known as calcification. Other segments of this system are teeth, , and all of the body. Periarticular tissue, also referred to as is situated at joints such as hip and knee and is firmly attached to the bone structure. It is difficult to separate it from the bones during dissection and therefore, sometimes becomes part of the skeletal weight and partly contributes to the variations observed in skeletal weights. Bone as a tissue represents an organic matrix the bulk of which is made up of the collagen. The inorganic matter consists mainly of deposits of calcium phosphate. On the other hand, bone as an organ comprises of red and yellow marrow, cartilage periosteum, blood and the bone tissue.

Based on the hardness, porosity and the content of present in them bone tissue is commonly divided into two compartments: compact (cortical) bone and trabecular (spongy and porous) bone. However, not all bones can be strictly classified as compact or trabecular since some types are intermediate in porosity and difficult to classify.

The compact bone is the hard dense part surrounding the outer walls of all bones, and it is predominant in the shafts of the long bones. Bone forming cells called the , synthesize the organic matrix (osteoid), tissue which undergoes mineralization. The trabecular bone is a spongy formation seen at the interior of flat bones and at the ends of long bones. It is highly porous (being soft and consisting mainly of bone marrow). It is also referred to as osseous tissue of the trabeculae, and if the matter also includes the soft tissue part then it is referred to as spongiosa.

Some investigators use the terminologies such as cancellous (spongy) bone and petrous (hard or the compact) bones.

In adults, typically 75-85 % of the total bone mass is considered to be compact bone and the rest trabecular bone [7,8]. After the skeleton has matured there is a continual net loss in trabecular bone mass, amounting to 25-45 % of the peak trabecular mass in normal human beings [9]. In the femur, there is considerably more bone loss with advancing age in trabecular than in cortical bone [10]. A typical long bone (femur, humerus) is a thick-walled, hollow, cylindrical shaft (the diaphysis) of compact bone. The central marrow cavity is the medulla. The ends of the shaft, mainly spongy bone covered by a thin cortex of compact bone, are called epiphysis. When actual growth is taking place, the epiphysis and the diaphysis are separated by columns of spongy bone (the metaphysis). Most bones are covered by the periosteum, a specialized connective tissue layer which can, if necessary, contribute to the formation of new bone.

The density of the skeleton is about 1.3 g/cc, and that of the dry mineralized collagenous bone matrix is close to 2.3 g/cc [11]. The mean density of vertebral cortical and trabecular bones has been reported to be 1.99 and 1.92 g/cc, respectively [12]. Densities reported for bones from Caucasian subjects aged 79 are as follow: fresh compact bone (from shafts of tibiae and femora) 1.85 g/cc, where as for fresh spongiosa (trabecular bone with marrow taken from thoracic and lumbar vertebrae and the calcaneus) was 1.08 g/cc [13]. These authors have also reported variations in densities between several subsamples taken from the same area of one bone. For example, the density of spongiosa (thoracic vertebrae) was found to be 0.8 to 1.4 g/cc, while densities of cortical bones (mid-schaft of the tibia) ranged from 1.5 to 2 g/cc.

The inorganic fraction is made of crystals forming hexagonal plates which are deposited in a regular way on and parallel to the axis of the collagen fibers. Bone crystals consist chiefly of , but they also contain carbonate, citrate and small amounts of sodium, magnesium, potassium, chloride, fluoride, and a number of trace elements. The crystals are surrounded by a hydration shell which allows free exchange of ions between extracellular fluid and the interior of crystals. Ions at the interior of the crystals have a slow turnover rate (some ions known as bone-seekers such as uranium, plutonium, lead, strontium and radium can be easily incorporated into the crystals).

The calcified mass of adult bone has three types of surface: a surface on which nothing seems to happen under normal physiological conditions (about 90 % of bone surface); a surface on which bone is being formed (controlled by osteoblasts, single nuclear cells); and a surface at which bone is being resorbed (bone resorption is controlled by multinuclear gain cells called ). Bone formation and bone resorption occur continuously and simultaneously throughout life, although the rates change with age and vary in different parts of the skeleton.

3.2 Representative bone samples

Homogeneity even within a particular type of bone samples is a much debated topic and it may be safely said that there are many diverse opinions. This is understandable because of the variations in proportions of the constituent compartments in a given bone segment, and the dynamics of bone growth.

The criterion for defining a particular bone sample as representative specimen mainly depends upon its usage and the associated logistics in obtaining the desired section of the bone. For example, in dealing with bone fracture cases, iliac crest will be biopsied for clinical studies since it can be done safely and easily. However, these biopsies serve as a proxy for those bones that suffer from fractures (e.g. osteoporosis), although iliac crest itself does not directly suffer from fractures. The argument here is that some one that has osteoporosis at one site is likely to have it generalized and that the iliac crest is probably representative. There is evidence that among various bones, iliac crest presents the best scenario of homogeneity [14]. On the other hand, bone is readily available in large amounts from subjects that fracture their hips and also from subjects undergoing hip replacement (e.g. arthritis). These situations offer other sampling possibilities for acquiring both normal and pathologic bone specimens.

The autopsy situation although beset with medico-legal barriers, when these barriers are overcome, offers the possibility of collecting rib, long bones, vertebrae etc, and have been the sources of bone samples. In many autopsy studies aimed at collecting skeletal data for radiological purposes, these sample have been frequently analyzed. Some times only a single type of specimen has been analyzed, and in some cases a combination of bones has been investigated.

In summary, with the exception of in vivo possibilities for assessment of total body Ca (and a few other elements) for diagnostic and other purposes (where facilities are available), despite severe limitations faced in obtaining biopsy samples from living subjects, this practice remains inevitable. Samples from autopsy cases, in spite of the fact that they too suffer from different type of limitations, provide the only practical possibility for undertaking comprehensive skeletal elemental concentration profile studies. Where possible, collection of multiple types of samples, analysis, and averaging the results for each skeletal segment would enhance the reliability of total skeletal content data.

3.3 Methods for sampling bone

Large quantities of bone samples are obtained at autopsy while limited quantities of biopsy specimens (especially as a part of a clinical procedure) are obtained by surgical intervention.

Biopsy sampling from living subjects require the usual sterile requirements and limitations on what type of gadgets can be used. Usually, a 8 mm diameter stainless steel trephine is used to obtain bone biopsy samples. The recovered quantity of bone permits separation of cortical and trabecular portions of bone by processing under clean bench conditions. Practical steps involved in such operations are presented in Figure 2 based on the procedure developed by Inskip et al [15]. Although this particular example is from a study designed to biopsy bones in monkeys, the technical aspects provide useful information for processing human samples.

Sampling at autopsy on the other hand, would permit use of conventional clean stainless steel equipment for removal of a section of bone from the body. After removal, samples are packaged in plastic bags and shipped under cool conditions (e.g. dry ice) to carry out the remaining part of the sampling operations that can be performed in the laboratory.

3.4 Methods for processing bone

Removal of connective tissues, soft tissues, fat and fibers is required. In some cases marrow part has to be separated. All these operations require considerable efforts and clean working conditions if the samples are intended for trace analysis.

Primary fragmentation of a chunk of bone can be accomplished by cooling the bone in liquid nitrogen and impacting it with a suitable implement. However, to avoid the risk of contamination and to provide a safe containment for the sample during the handling process, the sample should be enclosed in a Teflon bag and then cooled. Wedging the cooled bone between two plastic slabs (e.g. perspex blocks) and then impacting it with a stainless steel hammer would minimize such a contamination risk. This procedure yields small segments of samples and facilitates removal of marrow and other constituents, as well as partitioning the sample into subsections.

Distilled water, ether, acetone, chloroform, methanol, hydrogen peroxide and glacial acetic acid have been used separately or as mixtures have for defatting and cleaning. For cutting operations depending upon the purpose of the investigation, tools made of stainless steel and tungsten carbide can be used. It may even be possible to use tools made of quartz and titanium for finer operations or to remove soft parts of the bone thus minimizing exposure to hard metals. These tools can be cleaned with 1 % EDTA solution, followed by ultrasonic cleaning with demineralized water an methanol (10 %).

The processed samples should be preferably freeze-dried and stored under cool conditions until taken up for the measurement phase (see section 5.3). 4. TOOTH

4.1 The human teeth

The human adult has 32 permanent teeth. In each of the dental arcade there are two incisors, one canine, two premolar, and three molars.

In children, teeth are classified as deciduous or permanent. Deciduous teeth are formed during childhood and they are 20 in number: Incisors (central=4, lateral=4), canines =4, and molars (lst=4, 2nd=4). In addition there are 12 permanent: premolars=8 and 3rd molars=4.

The primary function of teeth is to mince food into small particles to pass through the esophagus.

4.2 Characteristics of tooth

The teeth consists of crowns that project above the gum, and single or multiple roots. Incisors have one root, lower and upper molars two and three roots, respectively.

The hard components of teeth are dentine, enamel, (specific gravity 3.0, 2.14 and 2.03, respectively), with dentine being the bulk component surrounding the chamber that contains the pulp. Dentine has no blood vessels or fibers. Those that line the inner surface of dentine, known as odontoblast, maintain the dentine deposits. The water content of dentine and cementum is about 10 % on a weight basis. Dentine also contains some firmly bound water. The pulp, being a soft tissue contains more water.

Enamel, which is the outer surface of tooth, is formed prior to eruption by epithelial cells called ameloblast. This is the hardest part in the body. Enamel is hydroxyapatite crystals embedded in fibers of keratin. Once the process of enamel formation is completed, no new enamel can be added. The water content of enamel is about 3 %.

The main inorganic constituents of both enamel and dentine are Ca, P, Mg and CO* Almost the entire weight of enamel (dry basis) is inorganic matrix, while that of dry dentine is 80 %.

Cementum, a bony substance is secreted by the periodontal membrane which lines the tooth alveolus (socket). Collagen fibers originating from the jaw bone pass through the periodontal membrane, making their way into cementum, and thus hold the tooth in place.

The pulp, which fills the tooth, consists of connective tissues, , blood vessels and lymphatics. The pulp is a soft tissue in composition and therefore has a higher water content than other compartments [16].

4.3 Methods for sampling tooth

A schematic diagram showing the components of tooth is shown in Figure 3. The sections includes whole tooth, tooth crown (enamel and dentine) and tooth roots (dentine).

Tooth as a whole entity is a heterogeneous matrix with two solid phases which represent different densities and also vary in their ability to concentrate trace elements. Therefore, sampling presents rather basic problems, and makes analysis of whole tooth less meaningful. The degree of variability of trace element distribution is further affected by increased sorption from decidual teeth and amalgam fillings (where applicable) in teeth from the adults. Deciduous teeth are obtained from dental clinics or schools or by approaching the parents directly. Occasionally, collection of healthy permanent tooth from living subjects is possible when tooth is extracted for orthodontal reasons. Hence sampling at autopsy is the only means of obtaining a complete set of samples that satisfy statistical design of a particular study. On the other hand, dental clinics are the best sources for pathologic specimens. Since the analysis of whole tooth is not meaningful, processing tooth samples further to extract the desired compartments is unavoidable and makes the analytical task rather tedious. The crown of tooth (enamel + dentine) is relatively easy to sample, but the relative proportions of the two tissues vary from specimen to specimen and the integrity of the specimen is compromised. Added to this, segments such as enamel are in such small proportions in a tooth matrix, sample contamination becomes a critical factor. Dentine presents special problems of contamination because of its porosity.

Hence the major problem with tooth is in the sample preparation step.

4.4 Methods for processing tooth

Tooth can be analyzed as a whole sample, or in sections: the sections are surface enamel, bulk enamel, bulk dentine circumpulpal dentine and cementum (enamel-dentine junction). Alternately, the whole tooth, tooth crown (enamel + dentine) and tooth roots (dentine) can also be analyzed. The sample treatment and processing steps differ depending upon the aim of the investigation. If the study is designed to investigate only the healthy teeth, those with fillings or caries should be discarded.

Removal of residual traces of blood, adhering soft tissues if any, and surface contamination will be required. Soaking in hydrogen peroxide, hypochlorite solution, acid wash and even brushing and scraping will facilitate removal of the residues. A detergent or acetone wash will accomplish removal of grease and fats. Mild scraping or brushing with appropriate tools and rinsing with high purity water is adequate if the teeth have not developed stains. Several procedures used in this connection have been reviewed [17,18].

Sectioning of teeth is required when whole tooth is not analyzed. Various cutting techniques have been used for isolating pulp, enamel and dentine: These include use of a diamond dental saw, dental burr drill, chipping and chiseling [17]. After the precleaning operations with distilled water and mild solvents, the crown (enamel + dentine) may be separated using the diamond saw. Further, enamel and dentine can also be separated, weighed and taken for further treatment such as dry or wet ashing.

Importantly, after the tooth is cleaned and being prepared for sectioning, the entire operation has to be carried out under clean conditions. The degree of difficulty posed by different sections during sample preparation is variable. Small quantities of samples obtainable from surface enamel make this part more prone to extraneous contamination. Analysis of bulk enamel, because of its abundance and low porosity characteristics can be expected to yield reliable results. Dentine on the other hand is a more porous fraction (hence easily contaminated) but it is regarded as a good indicator of lead exposure because of its growth characteristics [17].

The processed samples should be preferably freeze-dried and stored under cool conditions until taken up for the measurement phase (see section 5.3).

5. ANALYSIS

5.1 Analytical methods

A survey of the literature reveals that practically every available analytical technique has been applied to determine one or the other element in bones and teeth. The methods used are: atomic absorption spectroscopy (AAS), atomic emission spectroscopy (AES), anodic stripping voltametiy (ASV), mass spectrometiy (MS), X-ray methods particularly proton induced X-ray emission (PIXE) [19] and finally, nuclear activation techniques. All these techniques have been used in different modes depending upon the quantity of the sample available and elements sought. A review by Zwanziger [20] summarizes the applicability of a variety of methods for the analysis of bone. In a Canadian effort, a method based on AAS applicable to samples of a few mg has been demonstrated to be very effective [21].

Similarly, A review by Fergusson and Purchase [17] summarizes the applicability of a variety of methods for the analysis of tooth, with special reference to determination of lead.

Both destructive and non-destructive methods of analysis will be required to cover elemental analysis of a large number of minor and trace elements. Sample processing being a difficult operation for bone and tooth samples, non-destructive techniques such as neutron activation analysis (NAA) and X-ray based techniques such as X-ray fluorescence (XRF) and PIXE should be preferred. These techniques are suited for the determination of several elements in bone and tooth matrices.

Besides these in-vitro methods, for elements such as Ca in bone, in vivo approaches have also been utilized.

5.2. Reference materials for bone and teeth

There are only a few reference materials that are useful for quality control requirements of hard tissue analysis. The animal bone CRM (IAEA H-5) issued by the International Atomic Energy Agency (IAEA) is currently out of stock. The bone ash (NIST SRM 1400) and bone meal (NIST SRM 1486) issued by the National Institute of Standards and Technology are presently available. For tooth matrix, not even a single certified or recognized control material is presently available. 5.3 Preparation of bone and teeth for measurement

If the analysis is desired for whole bone and tooth, a few randomly selected small pieces of a particular type of bone or a tooth sample can be pulverized using the brittle fracture technique [22]. This procedure, which derives the benefit of making the samples brittle under liquid nitrogen cooling helps to pulverize the bone/tooth sample. The next treatment depends upon the method of analysis (destructive or non- destructive).

Obviously, the non destructive mode of analysis possible by techniques such as the NAA, PIXE, prompt gamma NAA and X-ray-Fluorescence is very attractive since it minimizes the work at the measurement stage of the analysis. It offers the possibility to investigate the variability of a given element on a population basis as large number of samples can be analyzed. Hence, in cases where these methods are applicable and are accessible, it is definitely of advantage to use them.

Much has been discussed in the literature about the presence of organic matter in bones as a source of interference during the measurement process and the need for removal of this fraction prior to analysis. However, if instrumental techniques are used then the samples may have to be merely encapsulated or pelletized and appropriate contamination control procedures have to be followed. On the other hand, if chemical intervention is required as is the case with some methods, the sample is either digested in high purity acids (wet ashing) or decomposed at low or high temperature depending upon the analyte in question (dry ashing). Both of these manipulations have their own short comings and the analyst's insight into these processes play a major role when results are reported.

Edward et al [23] have investigated the problem of dry ashing human and animal bone for a range of ashing times and temperatures. They have concluded that fluorine, chlorine, bromine, sodium, potassium and zinc were affected to varying degrees between 400 and 600 degrees C. For example, in the case of zinc it was shown that rat bone with biologically incorporated radioactive isotope (Zn-65) lost zinc after 6 hours of ashing and continued to lose up to 12 hours (i.e. even after the organic material had been removed). The loss was more pronounced at 500 and 600 °C treatments than at 400°C. Magnesium, calcium, strontium and manganese were unaffected. Zwanziger [20] has reviewed a number of ashing investigations related to several additional elements, and has pointed out in particular the poor reproducibility of results of lead analysis from ashed samples. Use of low temperature ashing aided by oxygen streams and us of Teflon containers for ashing have been shown to be safer, should dry ashing is the method of choice. Wet ashing aided by microwave oven appears to be the method of choice of many investigators. Dissolution in nitric acid is acceptable for most methods.

Concerning tooth, problems faced with dissolution are similar to that of bone. Teeth samples have been extensively analyzed in context of lead and several wet and dry ashing procedures are applicable if carefully carried out [17].

6. ELEMENTAL CONCENTRATIONS IN BONES

The elemental composition of bone surveyed as part of this review is presented in Table 3. It may be remarked that most publications do not satisfactorily describe the characteristics of the sampled bone as well as the methods adopted for preparation prior to analysis. Because of this lack of documentation it is very difficult to attribute whether the often observed differences are real or due simply to unspecified nature of the sample. For example, bone samples from aged subjects may have been in the subclinical state of osteoporosis but are classified as normal bone samples. Or a bone sample may contain small quantities of residual marrow and collagen. Under these conditions, analysis for a major element such as Ca would result in depressed concentration than what is likely to be normal. For the same reason, Fe results may tend to be high resulting from contributions due to the presence of even traces of red marrow. The reports were checked for analytical quality control (AQC) component, in particular to find out whether RM was used for validation of methods. This information is also shown in Table 3. It appears that rib samples are the frequently analyzed part of the skeleton.

6.1 Ca, N, O, P and minor elements

Oxygen; The oxygen component of the bone is reported to be between 30 and 46 %. Some fluctuation due to changes in moisture content is to be expected. Therefore, the distinction between fresh and dry weight basis also in not easy to identify.

Nitrogen: The nitrogen component of bone is reported to be between 4.4 to 5 %, with one exception which reported 12.2 %. The presence of soft tissue (e.g. marrow) and collagen can influence the nitrogen concentration and may account for high values.

Calcium: As expected, bone has been frequently analyzed for Ca. Besides activation techniques, especially NAA for both in vitro and in vivo determination of Ca, AAS and ICP-AES have also been applied. Typical concentration ranges reported are: cortical (20 to 22.5 %), trabecular (15.5 to 26 %) and whole bone (17 to 26 %). Occasional high and low values are also seen and it is difficult to interpret these results due to small number of samples involved, and methodological implications. No demonstrable link was observed between Ca concentration and geographical location of the subjects or between different types of bone samples. Similarly, the effects of age, sex, health, diet and metabolism were not readily apparent from the data, probably due to the underlying statistical reasons. Large groups of subjects need to be studied and compared. The number of studies reported from Japan is very impressive. Many groups have used available RMs. Phosphorus: Results from several countries obtained by different techniques show between 7 and 12.5 % between different types of bones, some based on fresh and some based on dry weight. The results overlap the stated range both on wet and dry basis and point to the need for establishing a strict protocol for analysis as well as for expressing the results. However, the average in many cases appear to fall within the narrow range of 10-12.5 % based only on dry weight. Some specific sections of the bone namely concha media and concha interior have been reported to contain P at concentrations exceeding 30 %.

Chlorine, Potassium, Sodium and Sulfur: Cl concentrations generally vary from 800 to 2700 mg/kg, including the spread from wet to dry basis, with one specific compartment showing as low as 200 mg/kg. Na falls in the range of 3000 to 8000 mg/kg on a dry weight basis. Very few results for K are available for comments, and there is a wide spread of concentrations reported ranging from about 50 to 6200 (>120 times) mg/kg. Sample characterization and analysis both need to be reviewed for this element before a reference value is assigned. For S, concentration of 800 mg/kg reported by one study raises concerns because of analytical problems (see section 7).

6.2 Trace elements in bone

Aluminum: ICP-AES has been frequently used by many investigators. Concentration of Al seems to be higher in ribs than in other areas of the skeleton. The results from Japan generally indicate high concentrations of Al. Al is believed to accumulate in bone with age [24]. Therefore, failure to document the age diminishes the credibility of the results if used for comparison. The over all range observed between various studies falls between 2 to 46 mg/kg. Among specific types of bone, a concentration of 19.5 mg/kg has been reported for iliac crest.

Antimony. Instrumental NAA is a good method for determining Sb if the concentration is not at the sub-ppb level. Very few results are reported for Sb in bone and the available findings suggest the average concentration to be in the low ppb range.

Arsenic: As is a difficult element to determine in biological samples, especially in bone. Therefore, it is not surprising that not many results have been reported for this element. Sample processing, e.g. dry ashing even at mild conditions can affect recovery. It appears that As is present at about 10 ppb or less.

Barium: Rather wide variations are seen in the results ranging from 1 to 35 mg/kg, when all the investigations are considered. However, of these a few selected investigations conform to a narrow range of 2.7 to 6 mg/kg, based on dry weight, providing an indication of probable average concentration of Ba in bone.

Boron: Very few results based on systematic studies have been reported for human bone. B is gaining recognition as a probable essential element for bone metabolism and therefore, there is a need for systematic studies of B in human bones, especially from different geographic locations and living conditions (e.g. vegetarians vs others). Like As, B is also susceptible to severe losses during processing and requires great care in sample preparation stages. The reported results range from 8 mg/kg on a fresh weight for one set of samples to 23 mg/kg for another set based on dry weight.

Bromine: Results available from half a dozen investigations suggest a range of 1.4 to 12 mg/kg as probable overall range (fresh or dry basis) without any clear separation.

Cadmium: AAS has been used by several investigators to determine Cd. A comparison of data spread over 6 countries does not reveal any particular trend since analytical problems are largely not resolved. Based mainly on AAS results, the best estimate for a majority of studies appear to fall in the range of 0.03 to 0.69 mg/kg (wet or dry, due to overlapping ranges). Cobalt: NAA and ICP-AES are well suited for the determination of Co and this is readily reflected in the results compiled. Based on dry bone, it appears that Co is present in bone at a concentration range of 0.015 to 0.13 mg/kg.

Chromium: Cr seems to be present in human bone at the ppm level. Since methods such as NAA permit non- destructive determination, in practice it is possible to minimize contamination. There appears to be a reasonable agreement between methods. With the exception of one study from China reporting divergent results, a concentration of 2.75 to 11 mg/kg appears to be the average range. Most of the samples are derived from ribs and both cortical and cancellous segments of bone have been investigated.

Cesium: NAA and ICP-MS are suitable methods for the determination of Cs. Results from Japan, show O.004 mg/kg by ICP-MS. Italian results based on dry weight indicate a range of 0.03 to 0.06 mg/kg, while the Chinese samples show a range of 0.05 to 0.08 mg/kg on ash weight basis. In fact the overall agreement is reasonable (ash weight is close to 50 % of dry weight), but work is needed to define a broad based reference range, since Cs is a radiologically sought element in bone.

Copper: Several analytical techniques have been used to determine Cu in bone. Analysis of bone for Cu involves the risk of contamination by reagents when ultra pure chemicals and water are not used during sample dissolution. Two studies using ICP-AES have reported a concentration exceeding 20 mg/kg (dry weight basis), while those using PIXE have reported between 5-19 mg/kg. Since these investigators did not use known RMs for AQC, the results seem to be open for discussion. An extensive investigation involving samples from USA reported 0.3 to 0.5 mg/kg (wet weight) in a variety of segments such as ribs, skull, tibia, vertebrae from both males and females. The remaining results are spread in the range of 1-5 mg/kg. Therefore, based on the current literature status, although reasonably good number of studies have been documented, it is not feasible to recommend a generalized reference value.

Fluorine: There are not many analytical methods (with the exception of ion-selective electrode) that are practical for the determination of F in biological materials. Nuclear based techniques are applicable only at high concentrations in specific matrices. This is reflected in very few RMs being certified for F in natural matrices. The overall range of concentration reported is 510 to 2100 mg/kg (dry weight). F is known to concentrate in bone, and this can explain the high values. The concentration of F in cortical (also referred to as compacta) is lower than in trabecular (also known as spongiosa) section. A difference, by a factor of 3 has been experimentally demonstrated for a bone sample taken from the distal epiphysis of a femur of a skeleton representing preindustrial populations [25]. hi high fluoride areas as in India, bone samples have been shown to contain extremely high concentration of F [26]. The F concentration in bone possibly increases with age. But this is difficult to ascertain because of the demonstrated inhomogeneity and wide variation in the distribution of F in bone along the length of a bone sample [25].

Iron: AAS, ICP-AES and NAA are most frequently used techniques. Fe concentration depends upon age, sex, diet and metabolism. There is a wide range of values reported for Fe for ribs. Part of the reason may stem from the reasoning presented under section 6. The effect of red (bone) marrow on the concentration of Fe found in bone can be minimized by careful handling of the sample. With the exception of one investigation from Japan, the remaining studies that have reported the results on wet or dry basis, indicate an overall range of 13 to 106 mg/kg. Results based on ash weight reported from China show high values.

Lead: AAS is the most popular method. Pb is one of the extensively studied elements because of the toxic implications. The risk of contamination during processing and loss during ashing is very high and requires good AQC. Concerning distribution of Pb in different sites of a particular type of bone, several differences have been observed in a study conducted on a skeleton belonging to preindustrial period [25]. The variations observed were, a factor of 2 for femur, 3 for tibia, 2.5 for fibula, 2 for ulna, and 3 for rib. Interestingly, for

10 the hip bone (i.e. iliac crest region) very little variation was observed from point to point, giving an indication that iliac crest (e.g. biopsy) maybe a good site for sampling. In Table 3, over 30 sets of results from several geographic regions are shown, and a majority of them fall in the range of 2 to 17 mg/kg irrespective of the basis (wet or dry) used to express the results. Practically every type of bone sample has been investigated. For individual bones, a restricted survey [27] states the following concentration as a general guide for analytical planning: rib 4-9, vertebra 2-8, femur 7-22, tibia 8-23 and skull 13-24, based on mg/kg dry weight. Bones samples from heavily industrialized and polluted areas generally tend to contain high concentrations of Pb in them as observed in samples from Poland (up to 70 mg/kg), (Table 3).

Magnesium: AAS, ICP-AES and NAA are applicable techniques. There appears to be not much difficulty in determining Mg in bone since the concentration range is high. A considerable number of studies have been reported for Mg in bone. The overall range appears to be between 0.17 and 0.37 %, with overlapping range for wet and dry weight based results. The samples are mainly from ribs, and include both cortical and trabecular segments.

Manganese: AAS, ICP-AES and NAA are most frequently used methods. The Mn concentration in bone appears to indicate geographical differences. There is a wide range of results reported by several investigators for various segments of the bone. It is difficult to arrive at a reference value. The overall range covers 0.14 to 8 mg/kg, irrespective of the basis used to express the concentrations. Mn is believed to be an important element in bone metabolism [28]. And several bone developmental effects have been linked to bone growth thus demonstrating the essentiality of Mn in bone metabolism [29].

Mercury: Like As and B, Hg may be easily lost if proper care is not taken during sample processing. Not many results are reported for this element. Being a nonessential element, the presence of Hg in tissues is an indicator of the exposure to this element through water, food or the conditions representing the environment, and it is difficult to arrive at a generalized reference value. Therefore, the limited results (0.018 to 0.62 mg/kg, dry weight basis) listed in Table 3. may be considered to indicate a particular level of exposure.

Molybdenum: There are only a few methods capable of determining Mo in biological materials at the level Mo occurs in these matrices. NAA is a very good method but very few analysts have access to such facilities. For these and other reasons there are only a few Mo results currently available for bone. Much work is warranted since dietary intake of Mo on a global basis is very different, and may be interesting to study its impact on bone metabolism.

Nickel: ICP-AES is the most frequently used method for determination of Ni in bone. A modest number of analytical results have been reported for this element. Excluding one high value (9.1 mg/kg) from a Japanese study, the remaining values fall in the range of 0.42 to 2.6 mg/kg.

Polonium: Po is a naturally radioactive element and is known as a bone seeker. By counting the radioactivity and making suitable corrections it is possible to assay minute quantities of Po in tissue samples. Results from two studies included in this compilation show Po concentrations well below part per trillion.

Rubidium: Instrumental NAA is particularly suited method for determining Rb in bone. Based on the available data, the average concentration may be somewhere in the range of 1 to 9.5 mg/kg. Of the 10 investigations included in this compilation, an Australian study that included bone marrow as part of the sample, has reported 25 mg/kg, while one from Russia without adequately defining sample and sampling characteristics has found 38 mg/kg.

Strontium: Nuclear analytical methods are very popular. Because of the close relationship with Ca metabolism in bone, Sr is an extensively investigated element and results are available from over 20 investigations. Not surprisingly, a wide range of results have been reported since bone is a target matrix for

11 Sr accumulation and factors such as intake and age influence the metabolism of Sr in bone. Thus a range of 50 to 420 mg/kg (wet or dry basis) seems to represent a plausible average profile. Unfortunately, many publications do not clearly indicate the geographic origin of the samples within the country Results reported from China and Russia are very high suggesting some environmental and nutritional factors. A clear identification of the bone area being analyzed, age of the subjects, and sample processing might help explain some of the extreme values reported in the literature. Sr is a radiologically sought element in bone.

Scandium: Instrumental NAA is particularly suited method for determining Sc in bone, and all the 4 sets of results included in this compilation are obtained by this method. The reported results cover a wide range from 0.001 to 0.19 mg/kg.

Selenium: Instrumental NAA is a good method for determining Se in bone. Based on 6 sets of results available from this review, 0.1 to 0.6 mg/kg appears to be the target range for reference values. More investigations are needed for confirmation.

Thorium: ICP-MS and RNAA are the chosen techniques for determining Th in biological matrices. In bone, INAA is also applicable if the concentration is not too low. Because of the limitations of choice of methods, not many investigations have been carried out to establish a broad based data base for Th in biological samples. Of the 2 investigations listed in this compilation, < 0.06 mg/kg has been reported for Japanese samples, and 0.2 to 0.4 mg/kg (ash basis) for samples from China.

Uranium: Radiochemical methods such as alpha spectrometry are the most sensitive methods for analysis of bones. Recently, ICP-MS has emerged as a powerful technique but the applications are just in the beginning stage. Results from Australia, USA, Russia, Nepal and China show average concentrations in the range of 0.001 to 0.062 mg/kg on an ash weight basis. Results for Japan are at the low end ranging from 0.0003 to 0.0009 mg/kg based on fresh weight of the tissue. Bone is a target organ for U, and is also a radiologically sought element.

Vanadium: Not many investigations are reported. ICP-AES is applicable, so is NAA if radiochemistry is used. The average concentration is probably in the range of 1-4 mg/kg, on a dry weight basis.

Zinc: AAS, ICP-AES, NAA, PIXE and XRF all are suitable for analyzing bone for Zn as reflected by numerous investigations. NAA is extremely well suited since it involves minimum sample preparation and purely instrumental determination. Over 25 sets of results for Zn have been scrutinized in this compilation. The average concentration ranges from 25 to 265 mg/kg, with overlapping ranges based on fresh, dry or ash weight. Zn concentrations in bone appear to depend on various factors such as nutrition, metabolism, and environment. The results also indicate inhomogeneous distribution of this element within the skeletal system. Considering only the U.S. results for rib, skull, tibia and vertebrae from both men and women, the range narrows to 25 - 58 mg/kg, based on fresh weight.

7. ELEMENTAL CONCENTRATIONS IN TEETH

The elemental composition of tooth surveyed as part of this review is presented in Table 4. As in the case of bone analysis, it should be recognized that most publications do not satisfactorily describe the characteristics of the sampled tooth as well as the methods adopted for preparation prior to analysis. The investigation by Cutress [30] studying the composition of the outer layer of dental enamel in samples obtained from 14 countries is a good example of the logistics involved in planning multi element studies. The results for Cu, Ba and S which appear to be extremely high have been traced back to possible methodological problem, in particular to disagreement with analysis of a reference sample. Further, the presence of caries if unnoticed compromises sample integrity. This is particularly important when teeth samples are sectioned

12 and in the absence of documentation it is very difficult to attribute whether the often observed differences are real or due simply to unspecified nature of the sampled fraction. For example, tooth samples may have been exposed to "filler" composites directly or as adjacent specimens. Or a tooth sample may contain small quantities of residual blood and soft tissue. Under these conditions, analysis for a major element such as Ca would result in depressed concentration than what is likely to be normal. For the same reason. The reports were checked for AQC, in particular to find out whether RM was used for validating the methods. This information is also shown in Table 4.

Methods applicable for bone are in principle also applicable to tooth analysis. Therefore, no reference to techniques will be made. Similarly, some of the general comments that can be seen under bone for selected trace elements are extendable to tooth, hence the comments under the following sections will be minimized.

7.1 Ca, P and minor elements

Unlike the large number of studies of Ca in bone, teeth appear to have been less widely investigated. The available results suggest the following concentrations: enamel from 31.5 to 37.3 %, dentine 31.5 % and whole tooth 10-12 % (samples from Polish subjects, appear to be low in concentration of Ca). However, individual investigations for dentine and whole tooth have been carried out less frequently than those for enamel. A link between dietary Ca, environmental factors and Ca content of tooth is not well established. However, more studies would be needed to confirm the relationship. Concerning P, the 5 studies listed in Table 4. show an average range of 16-21 % for enamel and 14 % for dentine. The results for Cl show a mixed picture. One set of results for enamel have been reported to be between 1600 to 2800 mg/kg while 2 other investigations indicate a range of 8000-9000 mg/kg. In dentine, the reported concentration is close to 2500 mg/kg, all based on dry weight. The concentration of K in enamel appears to differ widely between 190 to 1200 mg/kg, and in whole teeth one set of results are quoted to be just 53 mg/kg. Similarly, Na concentrations range from 1700 to 1300 mg/kg in enamel, 2000 to 2300 mg/kg in dentine and 1125 to 1193 mg/kg in whole tooth. Mg in enamel 745 to 4100 mg/kg, and in dentine around 1000-8000 mg/kg, based on dry weight. For S, results were available for only enamel showing an average concentration of 300 to 1350 mg/kg, with extreme values of 24 and 18780 mg/kg (all based on dry weight), for surface enamel. The high value appears to be due to analytical problem as the control sample has been stated to yield erratic results [30].

7.2 Trace elements

Aluminum: Results are available from another compilation for 14 countries and 3 other countries are listed in Table 4. The results, available only for enamel cover a wide range between 2 and 343 mg/kg. In some cases the basis used for expressing the results is uncertain and hence no definite comments can be offered for the presence of Al in teeth. None of the studies mentions the presence of caries in the teeth, and most groups exercise AQC. AQC check is specially needed in the case of Al since it is a difficult element to determine, and easy to introduce by way of contamination.

Barium: In view of the similarity of Ba with Ca and Sr, several investigations have dealt with these elements as a group. The overall range is spread between 2 and 22 mg/kg. Even the two studies reporting values identifying the basis (dry) for expression report 6.4 and 22 mg/kg. Hence a definitive statement on Ba in is not possible. Available RMs have been used for AQC.

Boron: Results are reported for enamel and the overall range is 1 to 8 mg/kg; Of these 2 investigations have mentioned the basis (dry) used, and these values are 4.1 to 5.3 mg/kg.

Bromine: Results are reported for enamel and the overall range is 1 to 4.5 mg/kg; Of these 1 investigation has mentioned the basis (dry), stating a concentration of 3.1 mg/kg for surface enamel.

13 Cadmium: Results have been reported for several countries. In Poland, Finland and the USA, samples from several geographical regions have been investigated. Available data suggest some dependence of the Cd concentration with geographical region in a given country. Enamel is the frequently studied fraction and the average concentration of Cd in this tissue appears to be in the range of 2 to 9 mg/kg, dry weight. The lowest concentrations of Cd were measured in dentine samples from Greenland and Denmark (0.086 to 0.097 mg/kg, dry) suggesting environment as a factor. Because of the difficulties in obtaining reliable results for Cd in biomatrices, the role of AQC has been well recognized in most of the studies. Interestingly, among the results tabulated in this compilation, studies that adopted strict AQC procedures reported low concentration of Cd in the samples.

Cobalt. The indicative concentrations for dentine and enamel appear to be 0.1 and 0.2 mg/kg, respectively. For whole tooth, the results appear to spread between 5 and 25 mg/kg. Sample characterization being poor, these results must be regarded carefully.

Chromium: Cr in tooth seems to depend upon a number of parameters. The first and the foremost is the AQC aspect. The various fractions show differing Cr concentrations depending upon the origin of the sample. Concentrations as high as 15 to 30, and 20 to 40 mg/kg (dry) have been reported for samples from Australia for dentine and enamel, respectively, while samples from a number of other countries can be grouped within a narrow range of 0.45 to 3.9 (dry) for both. On the other hand, whole tooth samples from urban and rural areas of Poland have been reported to contain as much as 42 to 47 mg/kg (dry) of Cr. Hence it is difficult to make any meaningful conclusions based on the available data.

Copper: The compilation includes samples representing many countries, and in several cases either multiple ethnic groups or differing geographic regions have been investigated. The results vary from country to country and indicate environment as a possible factor for the observed variations. Cu concentration of whole tooth (7 to 10 mg/kg, dry) generally exceeds that of dentine (0.8 to 5 mg/kg, dry?) or enamel (0.2 to 2.75 mg/kg, dry). Exceptional concentrations of 10 to 30, and 15-50 mg/kg (dry) have been reported for a set of enamel and dentine from Australia. Analytically, most investigators are capable of determining Cu without much difficulty and therefore, the observed variations seem to suggest real variations.

Fluorine: Determination of F in tooth should be scrutinized carefully, since methodologically it is still considered as an unresolved problem. The average concentration of F in enamel appears to fall in the wide range of 55 - 750 mg/kg. Tooth paste is a source of F, and careful documentation concerning dental hygiene practices of individuals is crucial. Therefore, it is difficult to conclude how much of this variation is originating from cosmetic sources, and how much is originating from diet and environmental sources, since all these are key players in the distribution of F in tissues. Some aspect of has discussed under F in bone (section 6.2).

Iron: There appears to be no clear separation of Fe in dentine, enamel or whole tooth. As seen from Table 4, dentine (3 to 30 mg/kg), enamel (15 - 138 mg/kg) and whole tooth (32 to 65 mg/kg), the only tendency seen relates to the low values seen for dentine. The use of AQC procedures is quite satisfactory. The link between concentration of Fe in tooth and age, diet, section of the tooth analyzed is unclear and needs further studies.

Mercury: Analysis of tooth samples for Hg are susceptible to multiple doubts. Sample preparation procedures may seriously hamper the status of Hg in the sample, so is the problem of Hg in the laboratory environment. In addition, Hg being part of the filler substance, case history of the sample has to be carefully scrutinized. The reported results do not indicate any specific tendency as such.

14 Magnesium: AAS and to some extent PIXE have been frequently used to determine Mg in tooth. Mg concentration in the enamel (specifically from the surface) is generally lower than that found in dentine. There appears to be no specific analytical problem in determining this element in tooth samples.

Manganese: There appears to be some tendency in the distribution of Mn in dentine, enamel and whole tooth. As seen from Table 4, dentine (1-4 mg/kg, except for Australian samples), enamel (0.1 to 2.0 mg/kg, except for Australian samples) and whole tooth (9 to 48 mg/kg). The only general discrepancy seems to be high in surface enamel as reviewed by Cutress [30]. The link, if any between concentration of Mn in tooth and age, diet, and the section of the tooth analyzed, is unclear and needs further studies. There have been some differences reported for tooth samples from different countries, and it may be due to geographical conditions. As discussed under Al, contamination problems for Mn are very severe, and extreme care should be exercised during analysis.

Molybdenum: Not many results have been reported for Mo. Also the results mostly concern the enamel fraction (0.1 to 2.4 mg/kg, dry), and hence no comments on specific trends can be provided.

Nickel: Nickel resembles Mn and Al in terms of problems faced during analysis, and careful attention to contamination control is a crucial requirement. Ni in enamel has been identified in the range of 0.4 to 1.2 mg/kg, with one value in the high range of 23 mg/kg in surface enamel. In whole tooth the concentration range appears to around 5 to 30 mg/kg. More data are required to reliably interpret the Ni concentrations in tooth and associated compartments.

Lead: Pb is a frequently investigated element in tooth. Many studies have been reported from several global regions. The results reported for urban areas are generally higher than those reported for non-urban environments. The following concentrations have been reported: enamel (2 to 150 mg/kg, dry, exceptions two sets of samples with 1100 and 1236 mg/kg for surface enamel), dentine (1 to 118 mg/kg, dry), and whole tooth (1 to 48 mg/kg dry, with one set showing 254 to 336 mg/kg on ash weight basis). Pb concentration in dentine tends to be high and reflects possible link to age, diet and other environmental factors. There is sufficient evidence to account for AQC by most investigators. One reason for this is the underlying toxicological importance of the data generated and the care taken during the planning stage of the investigation.

Strontium: Results are available from several countries. Very few results were found for whole tooth. On the other hand, almost every investigation studied the enamel and dentine fractions, with enamel receiving more attention in terms of number of studies. Analytically, Sr does not pose serious difficulties, and therefore comparative information is reliable. The following concentrations have been reported: enamel (82 to 204 mg/kg, dry, exception being some individual samples reported to contain concentrations up to 1400 mg/kg, and 7630 mg/kg for surface enamel), dentine (75 to 83 mg/kg, dry, with individual concentrations extending up to 250 mg/kg). A possible link appears to exist with geographical location but needs further confirmation.

Vanadium: Limited number of studies have addressed the problem of V determination in teeth. The enamel fraction has been analyzed by a couple of investigators and the range of concentrations reported lie between 0.003 to 0.02 mg/kg (with one exception reporting 1.2 mg/kg for surface enamel). The results reported for Yugoslavia are at the low end of the spectrum, suggesting that the study was conducted carefully with special attention to AQC. This study reports a concentration of 0.003 to 0.004 mg/kg in enamel (fresh weight?). On the contrary one study from Australia reports 10 to 35 mg/kg (for dentine and enamel). More data are needed to draw definitive conclusions.

Zinc: Zn in whole tooth appears to be in the range of 100 to 350 mg/kg. However, more results have been reported for dentine and enamel. Based on the results from several studies from varying geographical

15 locations, the probable average concentrations appear to range anywhere from 70 to 690, and 135 to 360 mg/kg, for enamel and dentine, respectively. Occasionally higher than the stated concentrations have been reported to both the fractions. It also seems that there is a general overlap of the concentration ranges irrespective of whether fresh or dry basis has been used for expressing the results. As discussed under bone, Zn values are likely to depend on various factors such as nutrition, metabolism and environment. Good AQC procedures have been applied by most investigators. Importantly, there is no specific analytical difficulty that interferes with the determination of this element in dental tissues as long as contamination from laboratory equipment and reagents is kept under surveillance.

8. DISCUSSION

8.1 Limitations of the data compiled

With the exception of a few investigations designed with great care to blend biological and analytical perceptions, many investigations that were reviewed during the course of this compilation suffer from a lack of interdisciplinary approach. A lack of anatomical background has frequently resulted in incompatible description of the samples taken for analysis. On the analytical front, to some extent the reliability of the results have suffered due to insufficient efforts to evaluate the methods for matrix suitability. Further, the limited availability of suitable RMs to validate the methods has also deteriorated the situation. Under these circumstances, it is not surprising that the observations resulting from several studies differ widely. Although an attempt is made in this compilation to identify the AQC component in a given investigation (see Tables 3 and 4), it should be recognized that these AQC observations apply mostly to the laboratory part of the work, and the fact that in many cases the sample validity itself is questionable indicates the basic nature of the problem.

For above mentioned reasons, it is emphasized that the results compiled in this report must be used with analytical caution and biological discretion.

8.2 Major and minor elements in bones and teeth

Bone as a matrix is rather complicated, and relating concentrations of major and minor elements to each other has to be done with great care. For example, when bones are properly sectioned, defatted, dried under standard working procedures and analyzed, certain postulations are valid: e.g. Ca and P are prime indicators of bone mineral, and N is the prime indicator of collagen and other . Under well defined conditions, Ca accounts for 25 % of dry fat-free bones [1]. From an analytical point of view, assessment of Ca after ashing at moderately high temperatures is not associated with any known problems. Wet ashing is perfectly safe for both Ca and P and many studies have reported a constant ratio for this element between different parts of a particular bone. Na, K, and Mg can be lost during ashing at high temperatures and thus part of the variation in the ratios is attributable to this factor. Wet ashing or cold temperature ashing is generally safe for most of the elements of this group. For S and to some extent also for K, very few studies were encountered during this review, and even these were of doubtful quality. Therefore, for these two elements, especially for S there is very little information in this compilation. The summary information is compiled in Table 1 (Bones) and 2 (Teeth).

8.3 Trace elements in bones and teeth

Trace element studies can be classified under two groups: Pb, Sr and Zn which are extensively investigated in bones in several countries, and others such as Al, Cd, Co, Cu, F, Fe, Mn, Rb and U. In tooth also, the tendency is similar. The summary information is compiled in Table 1 (Bones) and 2 (Teeth).

16 There are not many studies particularly looking into the distribution of trace elements within a particular human bone sample to localize the variations and seek a metabolic explanation, if any for such phenomenon. Braetter et al [25] have studied the distribution of Ca, F and Pb in one instance. Katie et al [31] have examined the temporal bone and have identified marked differences in the distribution of both structural (Ca, P and Mg) as well as the essential trace element Zn. They attribute these differences to the developmental specificities to be met by various regions of the bone and functional adaptation. In a Japanese study investigating sex and age related variations in elemental concentrations in rib samples, a total of 20 elements were studied [24]. Based on the analysis of rib samples from 28 male and 14 female subjects following conclusions were drawn: Bone Fe and Pb concentrations were significantly higher, while P concentrations were lower in males; elements Ca, P, Al, Fe, Mg, Na, Pb and Zn showed age related variations. Of these Al, Fe, Pb and Zn showed a linear relationship in accumulation with age.

In most countries Cd and Pb are considered to be priority pollutants, thus attracting more attention than rest of the trace elements. As can be seen from the number of studies on Pb in bones and teeth, extreme variations in concentrations are encountered. Obviously, living conditions greatly influence the accumulation of Pb. Age and Pb concentration in food, water and air, and the length of exposure are clearly linked to accumulation of Pb in bone and other tissues [32,33]. It is estimated that over 95 % of the Pb in the body at the age of 80 will be located in the bone [34], and therefore bone (i.e. skeleton) is considered as a reliable indicator of environmental exposure to Pb that can be measured in vivo by X-ray Fluorescence [35].

Grandjean and Jorgensen [18] have investigated the retention of Pb and Cd in prehistoric and modern human teeth. In 5000 years old samples of premolars from Nubia and 500 years old samples of teeth from subjects who had lived in Greenland, Pb concentrations were very low in comparison with modern specimens. For Cd, the results were reversed. The trend observed for Pb is explained by the excessive burdening of the environment with Pb, especially in the 70s and the 80s. The property of Pb to accumulate in teeth has served as a means to monitor lead exposure of human subjects, particularly the pediatric population.

Based on Cd in cancellous bone obtained from the iliac crest, it appears that the concentration is not related to age [36], but specimens from male subjects showed slightly higher concentrations of Cd than those from females. Interestingly, these investigators have also reported that the Cd content has a high statistically significant positive correlation with Sr and Ni in the same specimens. The toxicity of Cd to bone is documented through the autopsy studies of itai-itai disease cases [37] revealing reduction in bone density. Similarly, subjects living in Cd polluted environment have been shown to contain elevated levels of osteocalcin than non-exposed controls [38].

Toxicity induced by Al has become a concern. Although ingestion from food is low, prolonged exposure to food and water can result in the accumulation of Al in bone. Consequently, excessive accumulation of Al in the skeleton is believed to inhibit bone formation by interfering with normal bone metabolism [39]. The toxic effect of Al on the skeleton was recognized when epidemiologic link was established between incidence of dialysis encephalopathy and large amount of Al in the dialysate, and concomitant high incidence of fracturing osteomalacia [40]. The role of F in bone metabolism is well understood [26]. It has been extensively studied in connection with industrial and endemic fiuorosis as well as its role as a preventive medicine for bone and dental health.

The role of Zn in bone metabolism is being increasingly appreciated. The elevated concentrations of Zn in the urine of women with osteoporosis has been identified as a likely biomarker to screen post-menopausal women [41]. Saltman and Strause [34] advocate supplements of trace minerals for including Zn post- menopausal women to improve bone density. Cu is intricately linked to bone metabolism under both deficient and toxic conditions. Cu deficiency affects collagen metabolism while Cu toxicity is believed to induce depletion of bone density [42]. Mn is believed to be intricately linked with bone metabolism [28], and it is suggested that supplementation of Cu, Mn and Zn may add to the beneficial effect of Ca supplementation

17 by improving the bone mineral density in post-menopausal women [34]. Not much is known about Fe in bone metabolism. Boron is gaining importance as a beneficial element for bone metabolism. It is reported that supplementation of basal diet low in Mg and B with 3mg/d B significantly increased plasma concentration of ionized Ca in women thus establishing a mode of action for B in the pathway of bone metabolism [43].

9. CONCLUDING REMARKS

Chemical elements play a great role in the metabolism of bones and teeth. Some elements are beneficial (F at non toxic concentrations in bones and teeth, supplementation of Cu, Mn and Zn along with Ca to delay or prevent the onset of osteoporosis) and some others (chronic exposure to Pb even at moderate concentrations, and excessive exposures to F as in fluorosis situations) are detrimental for the normal functioning of the skeleton. Knowledge on the roles played by both groups of elements can be enhanced if reliable compositional picture is available for scrutiny.

The present survey was undertaken to assess the literature status on chemical composition of bones and teeth, and revealed that much needs to be done in order to have tangible collection of meaningful data. In this context, there is a desperate need for harmonization (types of samples chosen, procedures adopted to process the specimens, and finally the determination of analytes) to generate comparable data. To begin with, it is necessary to develop a bioanalytical protocol that exemplifies the merits and demerits of analyzing bones and teeth.

Identification of any particular type of bone as a representative sample for the whole skeleton appears to be a far cry. Even if such a representative segment of a particular bone is identified, the logistics related to medico-legal (autopsy) and anatomical (biopsy) parameters will prevail as decisive factors.

For the sake of gaining a comprehensive insight into the distribution of various trace elements in different types of bones, it is necessary to carry out controlled investigations on different types of bones (and cortical and trabecular segments from the same sources) from the same cadaver under well defined sampling conditions.

On the analytical side, development of hard tissue RMs for whole bone, as well as for cortical, trabecular and marrow segments separately, would be very helpful for future investigations.

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[16] EASTOE, J.E., The chemicalcomposition of teeth. In, Biochemist's Handbook, Long, C, (ed) D. Van Nostrand Co, New York, (1961), pp. 720-724.

19 [17] FERGUSSON, J.E., PURCHASE, N.G., The analysis and levels of lead in human teeth. Environ. Pollution 46(1987)11-44.

[18] GRANDJEAN, P., JORGENSEN P,I, Retention of lead and cadmium in prehistoric and modern human teeth. Environ. Res. 53 (1990) 6-15.

[19] CUA, F.T., HALL, G.S. Trace element analysis of human teeth and bone by proton Induced X-ray emission. Biol. Trace Ele. Res. 12(1987) 133-142.

[20] ZWANZIGER, H., The multi elemental analysis of bone: A review Biol. Trace Ele. Res. 19 (989) 195- 232.

[21] SUBRAMANIAN, K. S., CONNOR J.W., MERANGER, J.C., Bone lead analysis: Development of analytical methodology for milligram samples. Arch. Environ. Contain, and Toxicol. 24(1993)494- 497.

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[23] EDWARD, J.G, BENFER, RA., MORRIS, J.S. The effects of dry ashing on the Composition of human and animal bone. Biol. Trace Ele. Res. 25(1990) 219-231.

[24] YOSHINAGA, J., SUZUKI, T., MORITA, M. Sex and age related variation in elemental concentrations of contemporary Japanese ribs. Sci. Total Environ. 79(1989)209-221.

[25] BRAETTER, P., GAWLEK, D., ROESICK, U., A view into the past: Trace element analysis of human bone from former times. HOMO, Vol. 39 (1987), Book 2, pp 99-106.

[26] KRISHNAMACHARI, K.V.R.V., Fluoride. In, Trace Elements in Human and Animal Nutrition, (Mertz, W. Ed.) Academic Press, New York, Volume 1, 1986, pp. 365-407.

[27] SUBRAMANIAN, K. S. Lead. In, Quantitative Trace Analysis of Biological Materials, McKenzie, H.A., Smythe, L.E. eds.), Elsevier Publ, Amsterdam, 1988, pp. 589-603.

[28] STRAUSE, L., SALTMAN, P., Role of Manganese in bone metabolism. Chapter 5, In Nutritional Bioavailability of Manganese, (Kies, C, ed.), ACS Symp. Series 354, Washington D.C., 1987, pp. 46-55.

[29] HURLEY, L.L. Manganese, In, Trace Elements in Human and Animal Nutrition, (Mertz, W. Ed.) Academic Press, New York, Volume 1,1986, pp. 185-215.

[3 0] CUTRESS, T. W., A preliminary study of the micro element composition of the outer layer of dental enamel. Caries Res. 13(1979)73-79.

[31] KATIC, V., VUJICIC, G., IVANKOVIC, D., STAVLJENIC, A., VUKICEVIC, S., Distribution of structural and trace elements in human temporal bone. Biol. Trace Ele. Res. 29(1991)35-43.

[32] SAMUELS, E.R., MERANGER, J.C., TRACY, B.L., SUBRAMANIAN, K.S. Lead concentrations in human bones from the Canadian population. Sci. Total Environ. 89(1989)261-269.

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[34] SALTMAN, P., STRAUSE, L. The role of trace minerals in Osteoporosis. J. Am. College of Nutrition 4(1993)384-389.

[35] NORDBERG, G.F., MAHAFFEY, K.R., FOWLER, B.A., Lead in bone: Implications for dosimetry and toxicology. Environ. Health Persp. 91(1991)3-7.

[36] KNUUTTILA M., LAPPALAINEN, R., OLKKONEN H., LAMML S., ALHAVA, KM. Cadmium content of human cancellous bone. Arch. Environ. Health 37(1982)290-294.

[37] NODA, M., KITAGAWA, M., A quantitative study of iliac bone histopathology on 62 cases with itai- itai disease. Calcified Tissue International 47(1990)66-74.

[38] KIDO, T., HONDA, R., TSURITANI, I., ISHIZAKI, M., YAMADA, Y., NAKAGAWA, H., NOGAWA, K., DOHI, Y. Serum levels of bone Gla-protein In inhabitants exposed environmental Cd.. Arch. Environ. Health 46(1991)43-49.

[39] RODRIGUEZ, M., FELSENFELF, A. J., LLACH, F., Aluminum administration in the rat separately affects the and bone mineralization. J. Bone and Mineral Res. 5(1990)59-67.

[40] ALFREY, A.C., Aluminum. In, Trace Elements in Human and Animal Nutrition, (Mertz, W. Ed.) Academic Press, New York, Volume 2,1986, pp. 399-409.

[41] HERZBERG, M., FOLDES, J., STEINBERG, R., MENCZEL, J. Zinc excretion in osteoporotic women. J. Bone and Mineral Res. 5(1990)251-257.

[42] SAYMOUR, C.A., Copper toxicity in man. In Copper in Animals and Man. (MaC Howell, J., Gawthrone, J.M. Eds.)CRC Press, Boca Raton, FL, 198, pp. 79-106.

[43] NIELSEN, F.H., SHULER, T.R., ZIMMERMAN, T.J., UTHUS, E.O. Effect of boron depletion and repletion on blood indicators of Ca status in humans fed a magnesium low diet. J. Trace Ele. In Exp. Med., 3(1990)45-54.

21 ANNEXES (Data compilation)

22 Table 1: Range of Mean Values for Major, Minor and Trace Elements in Human Bones [yengar et al.,1978 (Pre-1978) Present Literature Values Sample Sample

Element Prep Range Prep Range Ag (mg/kg) a 1.1 a 0.041 - 0.061 Al (mg/kg) a 30 - 66.7 d 1.81-45.6 As (mg/kg) a 4.1 a 0.0011 Au (mg/kg) a <0.03 f <0.5 B (mg/kg) f 0.74 - 0.9 f 8 Ba (mg/kg) a 7.4 - 29 d 2.7 -5.93 Be (mg/kg) * < 0.0002 - < 0.001 Bi (mg/kg) a <0.2 Br (mg/kg) f 38 f 1.4 - 12.4 Ca(%) * 17-27.4 d 8.62 - 28.9 Cd (mg/kg) a 4.2 d 0.0247 - 2.2 Cl (mg/kg) f 632 d 228 - 2700 Co (mg/kg) * 0.01 - 0.029 d 0.0153-0.13 Cr (mg/kg) * 0.1-33 d 2.75 - 10.8 Cs (mg/kg) f 0.009 - 0.036 a 0.046 - 0.076 Cu (mg/kg) * 1.0-25.7 d 0.19-22.6 Eu (mg/kg) a 0.024 - 0.029 F (mg/kg) * 654-6180 d 639-2108 Fe (mg/kg) * 3-40 d 31.2-532.5 Hg (mg/kg) a <0.7 d 0.018 - 0.62 I (mg/kg) f 15 K (mg/kg) f 1470 d 47.9 - 6220 La (mg/kg) a <0.2 Li (mg/kg) d 0.23 Mg (%) * 0.070 - 0.098 d 0.01 -0.39 Mn (mg/kg) * 0.19-3 d 0.14-7.6 Mo (mg/kg) a 103 a 0.065 - 0.066 N (%) d 4.49 - 5.05 Na (%) * 0.560-1.41 d 0.316-0.805 Nb (mg/kg) * <0.07 Ni (mg/kg) a 110 f 0.42-9.12 O(%) f 30 -45.6 P(%) * 10.3 - 17.4 d 7.1 -12.5 Pb (mg/kg) f 10-42.5 d 0.57 - 70.66

23 Iyengar et al.,1978 (Pre-1978) Present Literature Values Sample Sample

Element Prep Range Prep Range Po (mg/kg) 12 12 (5.9-260)xl0" W (7.7 - 8.78) x lO' 9 9 Ra(mg/kg) a (10-12.5)xl0" W 1.09 xlO" Rb (mg/kg) * 0.1-5.11 d < 0.04 - 37.6 S (mg/kg) * 500 d 800 Sb (mg/kg) * 0.01-0.3 d 0.0151 Sc (mg/kg) f 4.6 d 0.0014 - 0.092 Se (mg/kg) * 1 - 8.95 d 0.13-0.56 Si (mg/kg) * 17 Sn (mg/kg) a 3.9 d 2.3 Sr (mg/kg) a 90.2 - 172 d 48.1-418 Ta (mg/kg) a 0.04 - 0.047 Tb (mg/kg) a 0.013 - 0.0344 Th (mg/kg) a 0.005 - < 0.04 a 0.24 - 0.38 Ti (mg/kg) d 1.95-< 5.0 Tl (mg/kg) f 0.002 U (mg/kg) a 0.0004 - 0.02 w 0.00055 - < 0.5 V (mg/kg) a 1.2 d <2.0-<6.0 W (mg/kg) * 0.00025 Y (mg/kg) * 0.07 Zn (mg/kg) * 50 - 170 d 91 -265.8 Zr (mg/kg) a <0.1 a 42.69 - 44.30

a = ashed d = dry weight f = freeze dried n = no sample preparation w = wet weight

* not clear or not addressed

24 Table 2: Range of Mean Values for Major, Minor and Trace Elements in Human Teeth Dentine Enamel Whole Tooth1 Sample Sample Sample Reference Prep Range Prep Range Prep Range

Iyengar et al.,1978 (Pre -1978) 0.004 -2.2 • 0.005 - 0.56 Present Literature Values 0.08 * 0.06-32

Iyengar et al.,1978 (Pre - 1978) 62 -136 12.5 - 86 Present Literature Values 2.1-343

Iyengar et al.,1978 (Pre -1978) 0.022 - 0.1 • < 0.02-0.07 Present Literature Values * <14 Att&Bg&g) Iyengar et al.,1978 (Pre - 1978) 0.03 - 0.07 <0.0001-0.11 Present Literature Values

Iyengar et al.,1978 (Pre -1978) * 5 Present Literature Values * 0.87 - 8.40 4.12

Iyengar et al.,1978 (Pre - 1978) * 129 * 4.2 -125 Present Literature Values * 2.02 - 22

Iyengar et al.,1978 (Pre -1978) * <0.01 Present Literature Values * 1.3-1.36

Iyengar et al.,1978 (Pre -1978) * 25 * 0.006 Present Literature Values * 0.001

Iyengar et al.,1978 (Pre - 1978) * 4.2-114 • 1.12-34 Present Literature Values * 3.1

Iyengar et al.,1978 (Pre -1978) * 25.0-28.2 * 36.0-37.4 Present Literature Values * 31.5 * 31.5-37.34 9.97-11.7

Iyengar et al.,1978 (Pre - 1978) * 0.099-0.12 * < 0.0001-0.51 Present Literature Values * 0.086 0.097 * 0.03-9.1 1.7-4.2 lyengar et al.,1978 (Pre -1978) 0.07

25 Dentine Enamel Whole Tooth1

Sample Sample Sample Reference Prep Range Prep Range Prep Range Present Literature Values 0.6 yengar et al.,1978 (Pre - 1978) 350-3900 • 3200 - 6500 Present Literature Values * 7900 - 8900

Iyengar et al.,1978 (Pre - 1978) * 0.006-1.11 * 0.004-0.13 Present Literature Values * 0.1 * 0.2 - < 16 4.8-24.8

Iyengar et al.,1978 (Pre - 1978) • 0.005-2 • 0.005 - 3.2 Present Literature Values * 0.45 - < 34 42.6-47.2

Iyengar et al.,1978 (Pre - 1978) * 0.04 Present Literature Values * 0.1

Iyengar et al.,1978 (Pre - 1978) * 0.21-28 * 0.26-33 Present Literature Values • 3 d 0.21-282 6.8 - 9.76

yengar et al.,1978 (Pre - 1978) <0.09 Present Literature Values

Iyengar et al.,1978 (Pre -1978) <0.04 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) 140- 157 * 293-2640 Present Literature Values * 54.3-752 lyengar et al.,1978 (Pre -1978) 31.7- 110 4.4 - 338 Present Literature Values * 27.95 -138 32.03-65

Iyengar et al.,1978 (Pre - 1978) <0.02 Present Literature Values * 6

Iyengar et al.,1978 (Pre -1978) < 0.08 Present Literature Values

Iyengar et al.,1978 (Pre -1978) * <0.02 Present Literature Values * 7.6

26 Dentine Enamel Whole Tooth1

Sample Sample Sample Reference Prep Range Prep Range Prep Range

Iyengar et al.,1978 (Pre - 1978) <0.08 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) <0.11-3.15 Present Literature Values 22 0.2

Iyengar et al.,1978 (Pre -1978) <0.02 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 670 0.036 Present Literature Values 0.05-2.04

Iyengar et al.,1978 (Pre -1978) <0.04 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) • 401 Present Literature Values • 191.04-1200 53.3 - 53.47

Iyengar et al.,1978 (Pre -1978) * <0.02 Present Literature Values * 1.4

Iyengar et al.,1978 (Pre - 1978) * 1.13 Present Literature Values * 0.53 -14

Iyengar et al.,1978 (Pre -1978) <0.02 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) * 6180-8700 * 1670-2800 Present Literature Values * 7926 * 745-4100

Iyengar et al.,1978 (Pre -1978) * 0.19 -10.5 * 0.28 - 30 Present Literature Values * 3.9 * 0.60 - 59 9.06-47.65

Iyengar et al.,1978 (Pre - 1978) * 2.3 * 0.054 -7.2 Present Literature Values * 0.1-2.37

27 Dentine Enamel Whole Tooth1 Sample Sample Sample Reference Prep Range Prep Range Prep Range Iyengar et al.,1978 (Pre - 1978) 5300-7500 * 3900-11600 Present Literature Values * 13600 1126-1193

Iyengar et al.,1978 (Pre -1978) 0.28 Present Literature Values 0.17

Iyengar et al.,1978 (Pre -1978) 0.045 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 1.35 Present Literature Values 0.9 0.29-23 4.6-31.3

Iyengar et al.,1978 (Pre - 1978) <0.09 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) 12.1-13.5 10.0-18.3 Present Literature Values 14 16.1-21

Iyengar et al.,1978 (Pre - 1978) 7.3 - 52 3.6 - 36 Present Literature Values 1.55-149.3 1.7-1236 1.65-53.5

Iyengar et al.,1978 (Pre -1978) <0.05 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 5.7 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 0.027 Present Literature Values 0.2

Iyengar et al.,1978 (Pre - 1978) 0.008 <90 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 9 - 86.5 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 0.39-73

28 Present Literature Values 0.15-4.61

Iyengar et al.,1978 (Pre - 1978) <0.04 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) <0.01 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) <0.04 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 500 - 670 48-281 Present Literature Values 24.19-18780

Iyengar et al.,1978 (Pre- 1978) 0.69 - 0.7 0.078 - 0.96 Present Literature Values 0.02- 8

Iyengar et al.,1978 (Pre -1978) <0.1 Present Literature Values

Iyengar et al.,1978 (Pre -1978) 12.8 0.27-0.872 Present Literature Values 1.47-18

Iyengar et al.,1978 (Pre -1978) 78-138 60 Present Literature Values 70

Iyengar et al.,1978 (Pre -1978) <0.08 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) 93 0.21 -120 Present Literature Values 1.60-< 325

Iyengar et al.,1978 (Pre -1978) 70 -100 81-111 Present Literature Values 75 -136.7 81.51-206.4 23

Iyengar et al.,1978 (Pre - 1978) Present Literature Values

Iyengar et al.,1978 (Pre - 1978) <0.02

29 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) 11-23 0.19 Present Literature Values 1.6-1.93

Iyengar et al.,1978 (Pre - 1978) 0.0047 <0.04 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) <0.02 Present Literature Values

Iyengar et al.,1978 (Pre - 1978) 0.007 - 0.034 Present Literature Values

Iyengar et al.,1978 (Pre -1978) < 0.01-0.017 Present Literature Values 0.0031 -1.4

Iyengar et al.,1978 (Pre -1978) 2.6 0.24 Present Literature Values <325

Iyengar et al.,1978 (Pre -1978) < 0.007 Present Literature Values 1.8

Iyengar et al.,1978 (Pre - 1978) Present Literature Values

Iyengar et al.,1978 (Pre -1978) * 173 -250 * 199-366 Present Literature Values * 135.1 -359.3 * 69.7 - 893 103-357

Iyengar et al.,1978 (Pre -1978) * 0.1 Present Literature Values * 0.08

1 = Whole tooth or regions not separated or specified d = dry weight basis n = no sample preparation

* not clear or not addressed at present

30 Table 3: Notations used in Bone Compilation a = ashed c = cortical d = dry weight cm — concha media f= freeze dried ci = concha inferior m = macerated cl = cancellous w = wet weight e = ethmoid

m = metatarsal

ma = maxilla

of= osfrontale

os = os parietale

pm = processus mastoides

r/u = radius/ulna

s = septum

t = trabecular

tb = tibia

tbf= tibia fragment

tbs = tibia section

te = temporal

AAS = atomic absorption spectrometry

C = colorimetiy

CPAA = charged particle activation analysis

EDX = energy-dipersive x-ray microanalysis

FT = fission track method

IGAA = instrumental gamma-activation analysis

ICPAES = inductively-coupled plasma atomic emission spectrometry

ICAP = inductively-coupled argon plasma

ICPMS = inductively-coupled plasma mass spectrometry

ISE = ion-selective electrode

LF = laser fluorimetry

MS = mass spectrometry

N = nitrogen analyzer

NAA = neutron activation analysis

PKE = proton induced x-ray emission

PIGE = proton induced gamma-ray emission

PGAA = prompt-gamma activation analysis

RAD = radiochemical methods

31 RBS = Rutherford backscattering spectrometry

XRF = x-ray fluorescence

V = voltametry

ICS = in-house calibration standard

L = no QA/QC mentioned but agreement with the literature values

L*= Lianqing and Guiyun, 1990 Z* = Zwanziger, 1989

32 Maior. Minor, and Trace Element in Human Bones (IAEA Compilation Post 19781

Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Bush etal., 1995 18-85 d 0.242 -14.21 (c) 1.81 ± 2.58 (c) 30 ICPMS (18M.12F) Yoshinagaetal., 1989 Japan 0.17-82 d 41.0 ±20.6 42 ICPAES (28M.14F) Kosugi et al., 1988 Japan d 45.6 x/, 1.3 18 ICPAES 11M.7F Mahanti and Barnes, 1983 a 42.9 ±0.8 3 ICPAES Gawlik etal., 1982 d 19.5 ±6.1 NAA Hyvonen-Dabek, 1981 24-70 f <22-54 <22 15 PIGE

Yoshingaetal., 1995 Japan d <0.4 35 ICPMS

Zaichick, 1994 d 0.47 ±0.20 NAA

Pietra et al., 1993 Italy w 0.0023-0.012 NAA Lin and Wen, 1988 China >10 a 0.061 ± 0.063 (M) 18, M NAA 0.041 ± 0.029 (F) 17 F

33 Age Sample Number of Analytical

Reference Location JTS. Preparation Range Mean Median Samples Technique

Yoshinga et al., 1995 Japan d <0.1-0.S <0.1 35 ICPMS

Pietraetal., 1993 Italy w < 0.002-0.010 NAA Mahanti and Bames, 1983 a 0.011 ±0.0005 3 ICPAES

Yoshinga etal., 1995 Japan d <0.1 35 ICPMS

Pietra et al., 1993 Italy w 0.00003-0.00039 NAA Huntinetal., 1982 f <0.5 10 V <0.5

Kosugi et al., 1988 Japan d 23.4 x/, 3.3 18 ICPAES 11M.7F Ward, 1987 New Zealand 69-84 d 19.79 ±4.18 14 PGAA

HyvSnen-Dabek, 1981 24-70 f <2-14 8.0 ±3.3 15 PIGE

Yoshinga et al., 1995 Japan d 0.41-3.20 1.3 35 ICPMS

34 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Kniewald et al., 1994 Croatia f 148.6-190.1 5 ICPAES Pietraetal., 1993 Italy w 1.7-3.4 NAA Samudralawar and Robertson, 1993 USA 60-82 f 36 ± 13 (c) 35.0 5 PKE 19±7(c/) 18 4 Manea-Krichtenetal., 1991 USA 67-96 d 3.53 -14.6 5.93 ± 4.37 6 (2M.4F) MS 2.82 -14.6 5.76 ±4.50 Zwanziger et al., 1987 & 1985 d? 17-314 NAA Jaworowski et al., 1985 France 34-89 d < 0.5 -15 2.7 ±3.9 22? ICPES Mahanti and Bames, 1983 a 21.0 ±0.4 3 ICPAES

Yoshinga et al., 1995 Japan d <0.2 35 ICPMS

Robertson et al., 1992 USA 60-82 f 1.1-12.8 (c) 4.1 ± 4.0 (c) 3.0 12 PKE 0.5-2.7(0 1.4 ±0.6(0 1.2 8M.4F Samudralawar and Robertson, 1993 USA 60-82 f 1.4±0.6(cQ 1.2 12 PKE 8M.4F Grynpas et al., 1987 d 110±55(c) 7 NAA 290 ±60 (del) 8 180±60(cQ 8 Smytheetal.,1982 Australia 3 - 80 (80%) d 10.3 ±4.5 7 XRF Hyvanen-Dabek et al., 1981 f 5.46 - 20.3 (n = 7) 12.4 ±5.5 15 PKE 9M.6F

35 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Bush etal., 1995 18-85 d 12.2-35.8 (c) 22.2 ± 4.4 (c) 30 ICPAES 30 (18 M, 12 F) Yoshingaetal., 1995 Japan 61-96 d 24.6 ±0.6 24.7 45 AAS 17M.28F & ICPAES Akesson etal., 1994 40-76 d? 25.8 ±0.7(0 5 EDX 22.2 ± 0.5 (0 3M.2F NAA 23.0 ±1.0(0 ICPES Kniewald etal., 1994 Croatia f 57.5-61.0 5 ICPAES Zaichick, 1994 d 22.6 ±2.1 NAA.XRF IGAA Samudralawar and Robertson, 1993 USA 60-82 f 19±2.2(cQ 19.0 12 PIXE 8M.4F Robertson et al., 1992 USA 60-82 f 12.6 - 27.5 (c) 20 ±4.1 (c) 21 12 PKE 14.8-22.2(0 19 ±2.4 (t) 19 8M,4F Katie etal., 1991 36-77 d 23.2-30.1 (fe) 26.7 ±2.3 (te) 174 ICPAES Manea-Krichten et al., 1991 USA 67-96 d 23.4-25.9 24.6 ±1.1 6 (2M, 4F) MS 22.3-25.4 24±1 Edward, 1990 65 w 23.5 ±0.2 1,M NAA Hisanaga et al., 1989 Japan 40-60 d 16.8 ±1.3 11 AAS Yoshinagaetal., 1989 Japan 0.17-82 r~~ d 23.9 ±0.9 24 42 AAS (28M.14F) Hisanaga et al., 1988 Japan 40-60 d 15.3 -18.8 16.8 ±1.3 11 AAS Kosugietal., 1988 Japan d 16.7 x/, 0.012 18 ICPAES 11M.7F Grynpas et al., 1987 d 22.5 ± 1.4 (c) 7 NAA

36 Age Sample Number of Analytical

Reference Location JTS, Preparation Range Mean Median Samples Technique 18.6 ± 2.8 (del) 8 15.5 ± 3.0 (et) 8 Igarashietal., 1987 Japan 37-94 w 14-19 17±2 6 ICPAES 3M.3F 4.1 -12 7.9 ±2.5 7 ICPAES 4M.3F 5.0 -13.0 8.7 ±2.7 7 ICPAES 4M.3F Zwanziger et al., 1987 & 1985 d? 6.4-28.7 NAA Jaworowski et al., 1985 France 34-89 d 7.3-22.6 18.5 ±3.4 22? ICPES Mahanti and Barnes, 1983 a 27.1 ±0.3 3 ICPAES

Niebergalletal., 1983 d? 29.4 3 ICAP Gawliketal., 1982 d 21.3 ±0.1 NAA Niese et al, 1982 d? 24.51 (op) NAA 25.53 (pm) 25.95 (ofi Smythe et al., 1982 Australia 3 - 80 (80%) d 8.62 ±2.52 7 XRF Hyvönen-Dabek, 1981 24-70 f 18.4-23.1 20.4 ±1.3 15 PIGE Lindh, 1981 65 d 25.58 IM PEXE Eichhorn et al., 1980 d? 28.9 ±11.5 NAA Giarratano et al, 1979 worf? 7.25 ± 1.06 1 NAA Hyvönen-Dabek, 1979 f 38.6 ± 0.7 (M) 8M RBS 37.5 ± 0.8 (F) 3F 37.1 ± 0.9 (M) 8M 37.8 ± 0.3 (F) 3F

37 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Bush etal., 1995 18-85 d 0-0.12(e) 0.029 ± 0.03 (c) 30 AAS 30 (18 M, 12 F) Baranowska et al, 1995 Poland 26-55 f 0.05 -1.5 0.69 ±0.38 25 (15 M, 10 F) AAS 0.05 - 0.38 0.14 ±0.10 10 (7M, 3 F) Yoshinga et al, 1995 Japan 61-96 d 0.28 ± 0.62 <0.1 45 AAS 17M.28F &ICPAES Kniewald etal, 1994 Croatia f > 0.001-0.053 5 V Pietra etal, 1993 Italy w 0.110-0.640 NAA Samudraiawar and Robertson, 1993 USA 60-82 f 2.7 ± 1.0 (c) 2.5 5 PKE 2.4 ± 1.2 (ct) 2.6 4 Saltzman et al, 1990 USA 20-74 w 0.07 ± 0.07 (M) 21 M AAS 0.11 ±0.13 (F) 5F 0.06 ± 0.06 (M) 21 M 0.08 ± 0.06 (F) 5F 0.05 ± 0.05 (M) 21 M 0.06 ± 0.03 (F) 5F 0.03 ± 0.04 (M) 21 M 0.03 ± 0.02 (F) 5F Hisanaga et al, 1989 Japan 40-60 d 0.57 ±0.26 11 AAS Yoshinagaetal, 1989 Japan 0.17-82 d 0.09 ±0.10 <0.1 18 ICPAES

Kosugi et al, 1988 Japan d 0.06 x/, 5.9 18 ICPAES 11M,7F Jaworowski et al, 1985 France 34-89 d 0.07 - 8.0 2.2 ±2.2 22? AAS Knuuttilaetal, 1982 m 0.24 ±0.17 (M) 88 AAS d 0.21 ± 0.16 (F) 61M.21F Liese, 1982 d? 0.0247 30 V

38 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Zaichick, 1994 d 1210 ±320 NAA

Edward, 1990 65 w 934.0 ±17.0 1,M NAA Grynpas et al., 1987 d 1300 ± 300 (c) 7 NAA 2700 ± 800 (del) 8 1600 ± 600 (cl) 8 Zwanziger et al., 1987 & 1985 d? 458-1197 NAA Nieseetal., 1982 d? 262 (op) NAA 356 iptri) 228 (of) Lindh, 1981 65 d 800 1M PKE Giarratano et al., 1979 worf? 2070 ± 665 1 NAA

Yoshingaetal., 1995 Japan 61-96 d 0.08 ±0.05 <0.1 45 AAS 17M.28F &ICPAES Zaichick, 1994 d 0.0153 ±0.0029 - NAA

Pietra et al., 1993 Italy w 0.044-0.073 NAA Yoshinaga et al, 1989 Japan 0.17-82 d 0.12 ±0.12 <0.1 8 ICPAES

Kosugi et al, 1988 Japan d 0.06 x/, 1.4 11(7M,4F) ICPAES 0.13 x/, 2.0 7(4M,3F)

39 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Lin and Wen, 1988 China >10 a 0.62 ± 0.52 (M) 23, M NAA 0.48 ± 0.46 (F) 24, F Zwanziger et al, 1987 & 1985 d? 0.48-3.84 NAA Gawliketal., 1982 d 0.046 ±0.037 NAA Mahanti and Barnes, 1983 a 0.4 ±0.01 3 ICPAES

Yoshinga et al, 1995 Japan 61-96 d <6.0 <6.0 45 AAS 17M.28F & ICPAES Zaichick, 1994 d 2.75 ±0.75 NAA

Pietra et al, 1993 Italy w 0.710-2.450 NAA Samudralawar and Robertson, 1993 USA 60-82 f 1.8 ± 0.7 {c) 3.0 5 PKE 1.9±0.8(cQ 3 4 Yoshinagaetal, 1989 Japan 0.17-82 d 4.87 ±1.14 4.88 38 ICPAES

Kosugietal, 1988 Japan d 10.8 x/, 1.1 18 ICPAES 11M,7F Lin and Wen, 1988 China >10 a 11.58 ±11.19 (M) 21, M NAA 5.72 ± 3.45 (F) 22, F Hyv6nen-Dabek et al, 1981 24-70 f 1.5-2.3(n = 3) <2.0 15 PDCE 9M.6F

Yoshinsaetal., 1995 Japan d < 0.004 35 ICPMS

40 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Pietra et al., 1993 My w 0.0295-0.056 NAA Lin and Wen, 1988 China >10 a 0.076 ± 0.048 (M) 17, M NAA 0.046 ± 0.013 (F) 13, F

Baranowskaetal., 1995 Poland 26-55 f 0.18-5.17 0.70 ±1.10 25 (15 M, 10 F) AAS 0.19-0.50 0.31 ±0.10 10 (7M, 3 F) Bushetal., 1995 18-85 d 0.5.0 (c) 1.0 ± 1.3 (c) 30 ICPAES 30 (18 M, 12 F) Yoshinga et al., 1995 Japan 61-96 d 0.19 ±0.20 <0.3 45 AAS 17M,28F & ICPAES Pietra etal., 1993 Italy w 0.790-2.148 NAA Samudralawar and Robertson, 1993 USA 60-82 f 6.3 ± 9.2 (c) 1.2 13 PIXE 1.4±4.1(cQ 0.7 13 Robertson et al, 1992 USA 60-82 f <0.7-32.3 (c) 5.1 ± 9.2 (c) 1.2 11 PDCE < 0.7 - 47 (t) 4.8 ±13(f) 0.7 6 Katie etal., 1991 36-77 d 14-30(fe) 20.5 ± 9.2 (fe) 77 ICPAES Saltzman et al., 1990 USA 20-74 w 0.41 ± 0.15 (M) 21 M AAS 0.44 ± 0.15 (F) 5F 0.34 ± 0.18 (M) 21 M 0.29 ± 0.18 (F) 5F 0.39 ± 0.22 (M) 21 M 0.52 ± 0.39 (F) 5F 0.38 ± 0.15 (M) 21 M 0.35 ± 0.07 (F) 5F Hisanaga et al, 1989 Japan 40-60 d 3.41 ±0.71 11 AAS

41 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Yoshinaga et al., 1989 Japan 0.17-82 d 0.23 ±0.22 <0.3 38 ICPAES

Kosugi et al, 1988 Japan d 22.6 x/, 1.1 11(7 M.4F) ICPAES 8.64 x/. 2.0 7(4M,3F) Jaksic et al, 1987 66 m 6 (ma) 3 XRF Hs) 30 4 5 (cm) 4 7 (ma) 3 PEXE 12 (e) 3 19 (ci) 4 16 (cm) 4 Mahanti and Barnes, 1983 a 1.0 ±0.03 3 ICPAES

Lappalainen et al, 1982 d? 1.3 ±0.5 138 AAS 87M.51F Smytheetal, 1982 Australia 3 - 80 (80%) d 1.5 ±0.8 3 XRF HyvSnen-Dabeketal, 1981 24-70 f 1.78-8.00 (n =14) 3.58 ±2.16 15 PKE 9M.6F »?£«)#&$ -

Yoshinga et al, 1995 Japan d < 0.01-0.06 <0.01 35 ICPMS

Yoshinga et al, 1995 Japan d < 0.01-0.04 <0.01 35 ICPMS

42 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Yoshinga et a!., 1995 Japan d < 0.008 35 ICPMS

Lin and Wen, 1988 China >10 a 0.029 ± 0.022 (M) 23, M NAA 0.024 ± 0.012 (F) 22, F

Zaichick, 1994 d 510±10 IGAA

Samudralawar and Robertson, 1993 USA 60-82 f 1710 ± 632 (cl) 1624 12 PKE 8M.4F Robertson et al., 1992 USA 60-82 f 1030-4360 (c) 2108 ± 940 (c) 1769 12 PIGE 731-3419(0 1947 ±741(0 1864 8\^4F Gawlik et al., 1982 d 626 ±573 NAA Hyvdnen-Dabek, 1981 24-70 f 178-1630 639 ±417 15 PIGE Suzuki et al., 1979 d 610 a 1100 d 530 a 960

Yoshinga etal., 1995 Japan 61-96 d 71.0 ±64.2 50 45 AAS 17M.28F &ICPAES

43 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Bush etal., 1995 18-85 d 9.0 -126 (c) 54±31.8(c) 30 ICPAES 30 (18 M, 12 F) Zaichick, 1994 d 59 ±17 NAA &XRF Pietra etal., 1993 Italy w 180-778 NAA Samudralawar and Robertson, 1993 USA 60-82 f 77±46(cQ 73 12 PKE 8M,4F Robertson et al., 1992 USA 60-82 f 5.1- 39 (c) 23±ll(c) 21 12 PKE 15-144(f) 71 ±45(0 73 8M.4F Katie etal., 1991 36-77 d 92 -124 (te) 105.6 ± 16.3 (te) 77 ICPAES Hisanaga et al., 1989 Japan 40-60 d 532.5 ±213.0 11 AAS Yoshinaga et al., 1989 Japan 0.17-82 d 31.2 ±41.3 16.3 34 ICPAES

Kosugi et al., 1988 Japan d 104 W1, 1.8 18 ICPAES 11M.7F Lin and Wen, 1988 China >10 a 1000 ± 1300 (M) 23, M NAA 630 ± 350 (F) 24, F Igarashi et al., 1987 Japan 37-94 w 8.6-35 23±9 6 ICPAES 3M,3F 94-410 240 ±140 7 ICPAES 4M.3F 1.9-26.0 13±9 7 ICPAES 4M,3F Jaksic etal., 1987 66 m 361 (ma) 3 XRF 360 (e) 3 374 (s) 30 624 (a) 4 642 (cm) 4

44 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique 690 (ma) 3 PKE 740 (e) 3 878 (ci) 4 934 (cm) 4 Zwanziger et al., 1987 & 1985 d? 50-345 NAA Mahanti and Barnes, 1983 a 29.4 ±0.6 3 ICPAES Gawliketal., 1982 d 183 ±78 NAA Hyv6nen-Dabek et al., 1981 24-70 f 4.97-10.4 7.58 ±1.55 15 PKE 9M,6F O'Connor et al., 1980 Australia 20-77 a 522 - 2164 1098 ±411 33 XRF 21M.12F

Yoshinga et al., 1995 Japan d <0.03 35 ICPMS

Yoshinga et al., 1995 Japan d < 0.008-0.2 < 0.008 35 ICPMS

45 Age Sample Number of Analytical Reference Location vrs. Preparation Range Mean Median Samples Technique

Yoshinga et al., 1995 Japan d <0.07 35 ICPMS

Yoshinga et al., 1995 Japan d < 0.2-0.6 <0.2 35 ICPMS

Bush etal., 1995 18-85 d 0-0.110(c) 0.018 ± 0.02 (c) 30 AAS 30 (18 M, 12 F) Zaichick, 1994 d 0.62 ± 0.29 NAA

Mahanti and Bames, 1983 a 0.012 ±0.0003 3 ICPAES

Yoshinga etal., 1995 Japan d < 0.004 35 ICPMS

Yoshinga et al., 1995 Japan d < 0.01-0.01 <0.01 35 ICPMS

Yoshinga etal., 1995 Japan 61-96 d 47.9 ± 15.2 43.5 45 AAS

46 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique 17M.28F &ICPAES Yoshinaga et al., 1989 Japan 0.17-82 d 80.7 ±58.2 62.8 42 AAS 28M.14F Kosugi et al., 1988 Japan d 429 x/, 1.9 11(7M,4F) AAS 87.2 x/, 1.6 7(4M,3F) Smytheetal., 1982 Australia 3 - 80 (80%) d 6220 ± 569 7 XRF Panday et al, 1980 d 4190 NAA w 1300

Yoshinga et al., 1995 Japan d < 0.3 -1.0 <0.3 35 ICPMS

Yoshinga et al., 1995 Japan d <0.05 35 ICPMS

Knuuttilaetal., 1982 d? 0.23 AAS t*J$ngftg>

Yoshinga et al., 1995 Japan d < 0.004 35 ICPMS

Pietraetal, 1993 Italy w 0.0049-0.014 NAA

47 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Bush etal., 1995 18-85 d 0.0582 - 0.3648 (c) 0.2219 ± 0.0698 (c) 30 ICPAES 30 (18 M, 12 F) Yoshinga et al., 1995 Japan 61-96 d 0.285 ± 0.020 0.284 45 AAS 17 M, 28 F & ICPAES Akessonetal, 1994 40-76 d? 0.31 ± 0.02 (f) 5 EDX 0.26 ±0.02(f) 3M.2F NAA Zaichick, 1994 d 0.276 ±0.013 NAA &IGAA Samudralawar and Robertson, 1993 USA 60-82 f 0.270 ± 0.040 (ct) 0.270 12 PIGE 8M,4F Robertson et al., 1992 USA 60-82 f 0.17- 0.33 (c) 0.26 ± 0.05 (c) 0.25 12 PIGE 0.18-0.34(r) 0.27 ±0.04(0 0.27 8M,4F Katie etal., 1991 36-77 d 0.254-0.291 (fe) 0.272 ± 0.018 (te) 174 ICPAES Edward, 1990 65 w 0.2236 ±0.0190 1,M NAA Yoshinaga et al., 1989 Japan 0.17-82 d 0.28 ±0.03 42 ICPAES 28M.14F Kosugi etal., 1988 Japan d 0.1830 W, 0.0001 18 ICPAES 11M,7F Grynpas et al., 1987 d 0.22 ± 0.02 (c) 7 NAA 0.20 ± 0.03 (del) 8 0.17±0.03(c/) 8 Zwanziger et al., 1987 & 1985 d? 0.160-0.550 NAA Jaworowski et al., 1985 France 34-89 d 0.1230-0.3015 0.246 ±0.035 22? ICPES Mahanti and Barnes, 1983 a 0.226 ±0.004 3 ICPAES

Niebergall et al., 1983 d? 0.31 3 ICAP Niese et al., 1982 d? 0.369 (op) NAA

48 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique 0.260 (pm) 0.309 (of) Hyvonen-Dabek, 1981 24-70 f 0.1580-0.2550 0.2078 ±0.029 15 PIGE Lindh, 1981 65 d 0.39 1M PKE Panday et al., 1980 d 0.01 NAA w 0.031

Bush etal., 1995 18-85 d 0.06-0.29 (c) 0.14 ± 0.06 (c) 30 AAS (18M,12F) Yoshingaetal., 1995 Japan 61-96 d <3.0 <3.0 45 AAS 17M.28F & ICPAES Samudralawar and Robertson, 1993 USA 60-82 f 1.2 ± 0.2 (c) 1.20 5 PKE 1.01 ± 0.4 (ct) 1.20 8 Hisanaga et al., 1989 Japan 40-60 d 6.8 ±1.6 11 AAS Yoshinaga et al., 1989 Japan 0.17-82 d <2.5 0 ICPAES

Kosugi et al., 1988 Japan d 5.32 x/, 1.9 11(7 M.4F) ICPAES <1.0 6 Zwanziger et al., 1987 & 1985 d? 0.32 -1.36 NAA Mahanti and Barnes, 1983 a 1.56 ±0.08 3 ICPAES Niebergall etal., 1983 d? 6.2 3 ICAP Niese etal., 1982 d? 7.6 (op) NAA 3.0 (pm) 3-5 (of) HyvSnen-Dabeketal., 1981 24-70 f 1.5-3.6(n = 6) <2.3 15 PKE 9M.6F

49 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Stein et al., 1979 d? 5.7 ±2.2 AAS 2.3 ± 0.6 2.8 ±2.1

Yoshinga etal., 1995 Japan d < 0.08-0.1 <0.08 35 ICPMS

Lin and Wen, 1988 China >10 a 0.066 ± 0.039 (M) ll.M NAA 0.065 ± 0.048 (F) 8.F Jaksic et al., 1987 66 m 30 (e) 3 PDCE 21 (a) 4 11 (cm) 4

*<*}

Zaichick, 1994 d 5.05 ±0.26 NAA &IGAA Samudralawar and Robertson, 1993 USA 60-82 f 4.8 ± 0.6 (c) 4.6 12 PIGE 4.4 ± 0.6 (c) 4.2 8M,4F Yoshinaga etal., 1989 Japan 0.17-82 d 4.92 ±0.18 42 N 28M.14F HyvSnen-Dabek, 1981 24-70 f 11.4-13.0 12.2 ±0.8 15 PIGE Lindh, 1981 65 d 4.49 1M PKE

Yoshinga etal., 1995 Japan 61-96 d 0.518 ±0.046 0.514 45 AAS

50 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique 17 M, 28 F &ICPAES Zaichick, 1994 d 0.638 ± 0.055 NAA

Samudralawar and Robertson, 1993 USA. 60-82 f 0.540 ± 0.060 (ct) 0.520 12 PIGE 8M.4F Robertson etal., 1992 USA 60-82 f 0.350 - 0.640 (c) 0.520 ±0.080 (c) 0.510 12 PIGE 0.460-0.610(0 0.520 ± 0.060 (f) 0.510 8M.4F Edward, 1990 65 w 0.4961 ±0.017 l.M NAA Yoshinaga et al., 1989 Japan 0.17-82 d 0.523 ±0.988 0.54 42 ICPAES 28M.14F Kosugietal., 1988 Japan d 0.3880 x/, 0.0001 11(7M,4F) ICPAES 0.3160 x/, 0.00012 7(4M,3F) Grynpas et al., 1987 d 0.60 ± 0.03 (c) 7 NAA 0.62 ± 0.09 (del) 8 0.52 ± 0.14 (ct) 8 Zwanziger et al., 1987 & 1985 d? 0.350-0.740 NAA Mahanti and Barnes, 1983 a 1.066 ±0.02 3 ICPAES Gawlik et al., 1982 d 0.490 ±0.090 NAA Niese etal., 1982 d? 1.401 (op) NAA 1.309 (pm) 1.476 (of) Hyvonen-Dabek, 1981 24-70 f 0.5230-0.6720 0.5763 ±0.0371 15 PIGE Lindh, 1981 65 d 0.58 1M PKE Pandayetal., 1980 d 0.805 NAA w 0.25

51 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Yoshinga et al., 1995 Japan d <0.1 35 ICPMS

Yoshinga et al., 1995 Japan d < 0.02-1.0 <0.02 35 ICPMS

Baranowska et al., 1995 Poland 26-55 f 0.14-2.58 0.38 ±0.55 25 (15 M, 10 F) AAS 0.14-0.60 0.26 ±0.15 10 (7M, 3 F) Yoshinga etal., 1995 Japan 61-96 d <1.5 45 AAS 17M.28F &ICPAES Samudralawar and Robertson, 1993 USA 60-82 f 2.6±2.1(c) 1.50 10 PKE 2.3 ±U(cQ 1.40 10 Robertson et al., 1992 USA 60-82 f < 1.2- 7.8 (c) 1.8 ± 2.1 (c) 1.5 6 PKE < 1.2-4.9(f) 1.6 ±1.3(0 1.2 7 Hisanaga et al, 1989 Japan 40-60 d 9.12 ±1.82 11 AAS Yoshinaga etal, 1989 Japan 0.17-82 d 0.42 ±0.40 <0.4 40 ICPAES

Kosugi et al, 1988 Japan d 0.32 x/. 1.7 18 ICPAES 11M,7F Mahanti and Barnes, 1983 a 2.1 ±0.02 3 ICPAES

Knuuttila etal, 1982 d? 1.29 AAS Hyvdnen-Dabek et al, 1981 24-70 f 1.9-3.0(n = 4) <2.4 15 PKE 9M.6F

52 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Samudralawar and Robertson, 1993 USA 60-82 f 30 ± 7 (cQ 33.0 12 PIGE 8M.4F Robertson et al., 1992 USA 60-82 f 6-35(c) 30 ± 8.2 (c) 32 12 PIGE 26-38(0 33 ±3.8(0 33 8M.4F Hyvdnen-Dabek, 1981 24-70 f 30.4-38.1 34.8 ± 2.3 15 PIGE Lindh, 1981 65 d 40.52 1M PIXE HyvSnen-Dabek, 1979 f 44.8 ± 0.8 (M) 8M RBS 45.0 ± 0.9 (F) 3F 45.6 ± 1.2 (M) 8M 45.1 ± 1.2 (F) 3F

Yoshinga et al., 1995 Japan 61-96 d 11.9 ±4.0 120 45 AAS 17 M, 28 F & ICPAES Akesson et al., 1994 40-76 d? 10.5 ±0.1(0 5 EDX 9.8 ±0.2(0 3M,2F NAA 10.0 ±0.4(0 ICPES Zaichick, 1994 d 10.9 ±0.7 NAA &IGAA Samudralawar and Robertson, 1993 USA 60-82 f 9.6 ± 1.3 (cQ 73 12 PIGE 8M.4F Robertson etal., 1992 USA 60-82 f 4.1- 11.2 (c) 8.8 ± 2.1 (c) 9.5 12 PIGE 6.5-11.1(0 9.4 ±1.3(0 9.8 8M.4F

53 Age Sample Number of Analytical

Reference Location JTS. Preparation Range Mean Median Samples Technique Katie etal., 1991 36-77 d 10.1-12.9 (re) 11.5 ± 1.0 (te) 174 ICPAES Yoshinaga et al., 1989 Japan 0.17-82 d 12.3 ±0.40 42 ICPAES 28M.14F Kosugi et al., 1988 Japan d 8.02 x/, 0.12 18 ICPAES 11M.7F Giynpas et al., 1987 d 9.7 ± 0.5 (c) 7 NAA 8.7 ±1.1 (del) 8 7.1±1.4(cQ 8 Jaksic et al., 1987 66 m 29 (ma) 3 XRF 28 (e) 3 34(5) 30 54 (a) 4 50 (cm) 4 10 (ma) 3 PKE 22 (e) 3 26 («•) 4 31 (cm) 4 Mahanti and Bames, 1983 a 13.8 ±0.2 3 ICPAES Niebergalletal., 1983 d? 12.5 3 ICAP Gawlik etal., 1982 d 9.8 ±0.6 NAA Hyvonen-Dabek, 1981 24-70 f 8.42 -10.5 20.4 ±1.3 15 PIGE Lindh, 1981 65 d 12.19 1M PKE Eichhom etal., 1980 d? 10.4 ±4.1 NAA Hyvonen-Dabek, 1979 f 16.6 ± 0.5 (M) 8M RBS 17.4 ± 0.8 (F) 3F 17.3 ± 0.5 (M) 8M 17.1 ± 0.9 (F) 3F

54 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Baranowska et al., 1995 Poland 26-55 f 2.90 -204.53 70.66 ±52.58 25 (15 M, 10 F) AAS 8.90-40.10 24.76 ±11.80 10(7M,3F) Bush etal., 1995 18-85 d 0.23-6.45 (c) 2.2 ± 1.5 (c) 30 AAS 30 (18 M, 12 F) Deibeletal., 1995 USA f 1.4-11.5(0 5.0 ±3.9(0 6 PKE 1.3-45 (c) 12.6 ± 12.9 (c) 6 PKE 1.54-11.75 6.55 ±2.93 7 AAS Yoshingaetal., 1995 Japan 61-96 d 6.85 ±3.41 5.55 45 AAS 17M,28F &ICPAES Kniewald etal., 1994 Croatia f 1.06-5.80 5 V Samudralawar and Robertson, 1993 USA 60-82 f 5.0 ± 3.3 (c/) 3.80 12 PKE 8M,4F Keinonen, 1992 Finland 13-64 1.8-3.7 15 M 13M,2F Robertson etal., 1992 USA 60-82 f 5.4-45 (c) 12.3 ± 10.9 (c) 9.1 12 PKE < 1.5-11.5(0 4.9 ±3.4(0 3.8 8M,4F Manea-Krichten et al., 1991 USA 67-96 d 6.73-23.4 12.8 ±6.0 6 (2M, 4F) MS 4.4-14.3 7.18 ±3.65 Erkilla etal., 1990 Finland 35.5 ±8.9 w 4.6±11.3(<6) 22 InXRF 33.1 ±31.7 (r/u) Hu etal, 1990 50-91 a 26.7 ± 9.7 (c) 27 AAS 24.4 ± 11.1 (t) 15M.12F Morgan etal., 1990 England w 10-42 7.2 ±10.8 59 InXRF Saltzmanetal., 1990 USA 20-74 w 6.70 ± 3.96 (M) 46M AAS 3.17 ± 0.90 (F) 8F 12.46 ± 8.73 (M) 46M

55 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique 6.96 ± 2.55 (F) 8F 12.55 ± 10.65 (M) 46 M 4.54± 2.04 (F) 8F 4.12 ± 2.49 (M) 46 M 2.01± 0.72 (F) 8F Hisanaga et al., 1989 Japan 40-60 d 4.47 ±1.98 11 AAS Kowaletal., 1989 Canada 55 d 18-50 29.8 ± 13 AAS Samuels et al., 1989 Canada 3 20 a 14.77 ±1.30 25 AAS Yoshinaga et al., 1989 Japan 0.17-82 d 3.26 ± 2.42 2.88 30 ICPAES

Hisanga et al., 1988 Japan 40-60 d 1.94-8.15 4.47 ±1.98 11 AAS Kosugi et al., 1988 Japan d 0.57 x/, 3.1 18 ICPAES 11M.7F Somervaille et al., 1988 England 49.3 w 9.4 (tb) 20 InXRF 12M,8F Aalbers et al. 1987 Netherlands 16-80 d 1.0-19 5.0 ±3.2 4.2 189 (130 M, 70 F) AAS Valkovic et al., 1987 2-75 m 11-570 (x) 9 XRF 13-370 (cm) 6M.3F 11- 840 (ci) Somervaille et al., 1986 a 22.3 -65.6 (ml) 3 XRF 17.7- 55.6 (m2) 3 17.2 - 83.0 (tbs) 21 6.5- 42.5 (tbf) 11 22.1 -73.5 (ml) 3 AAS 19.9- 56.8 (ml) 3 15.1- 79.2 (tbs) 21 10.0- 60.3 (tbf) 11 Jaworowski et al., 1985 France 34-89 d 5.0-35 16.9 ±10.1 22? AAS

56 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Mahanti and Barnes, 1983 a 22.4 ±0.4 3 ICPAES

Wielopolski etal., 1983 w 15-35(r) AAS Liese, 1982 d? 7.7 15 V HyvSnen-Dabeketal., 1981 24-70 f 9.01 • 19.0 12.2 ±2.5 15 PIXE 9M,6F Lindh, 1981 Sweden 10-51 3 PKE 10-34 3 Wittmers et al., 1981 a 14.08 ±1.74 AAS 60.85 ± 5.24 O'Connor et al., 1980 Australia 20-77 a 14-253 47 ±53 33 XRF 21M.12F

Grandjean et al., 1979 Denmark 16-54 5.5 17 AAS 9M.8F Emslander, 1978 d? 1.8 - 7.7 4.9 ±1.5 20 AAS

Takizawa etal., 1990 Japan 45-83 w 7.7 ± 2.2 22 RAD (16M.6F) Henshaw et al., 1987 United Kingdom 22-82 w 8.78 ± 0.78 4 RAD (2 M, 2 F)

Yoshingaetal., 1995 Japan d < 0.01-0.3 <0.01 35 ICPMS

57 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Yoshinga et al., 1995 Japan d <0.06 35 ICPMS

Henshawetal., 1987 United Kingdom 22-82 w 1.09 ± 0.38 4 RAD (2 M, 2 F)

Yoshinga et al., 1995 Japan d <0.08 35 ICPMS

Pietra et al., 1993 Italy w 3.29-6.45 NAA Zaichick, 1994 d 37.6 ±5.2 NAA

Samudralawar and Robertson, 1993 USA 60-82 f 2.1 ± 3.0 (c) 1.0 11 PDCE 2.1 ±3.1 (ct) 1.0 12 Robertson et al., 1992 USA 60-82 f < 0.6- 1.8 (c) 1.0 ± 0.5 (c) 1.0 9 PKE < 0.6 -2.8(f) 1.1 ±0.8(0 1.0 9 Lin and Wen, 1988 China >10 a 6.56 ± 3.21 (M) 23, M NAA 5.32 ± 3.06 (F) 22, F Zwanziger et al., 1987 & 1985 d? 2-62 NAA Gawliketal., 1982 d <0.04 NAA Smytheetai., 1982 Australia 3 - 80 (80%) d 9.5 ±2.8 7 XRF

58 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique O'Connor et al, 1980 Australia 20-77 a 17-42 25 ±7 33 XRF 21M,12 F

Yoshinga et al, 1995 Japan d < 0.004 35 ICPMS

Lindh, 1981 65. d 0.08 1M PKE

Yoshinga etal, 1995 Japan d <0.1 35 ICPMS

Zaichick, 1994 d 0.0151 ±0.0032 NAA

Pietra et al, 1993 Italy w < 0.0015-0.00715 NAA Zwanziger et al, 1987 & 1985 d? 0.004-0.081 NAA

Zaichick, 1994 d 0.092 ±0.79 NAA

Pietra etal, 1993 Italy w 0.00005-0.0008 NAA

59 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique Gawlik et al., 1982 d 0.0014 ±0.0007 NAA Lin and Wen, 1988 China >10 a 0.19 ± 0.20 (M) 23, M NAA 0.12 ± 0.086 (F) 24, F

Yoshinga et al., 1995 Japan d <2 35 ICPMS

Zaichick, 1994 d 0.226 ±0.073 NAA

Piefra et al., 1993 Italy w 0.150-0.480 NAA Mahanti and Barnes, 1983 a 0.101 ±0.002 3 ICPAES Gawlik et al., 1982 d 0.13 ±0.4 NAA Smythe et al., 1982 Australia 3 - 80 (80%) d 0.56 ±0.19 2 XRF

Yoshinga et al., 1995 Japan d < 0.004-0.2 < 0.004 35 ICPMS

Yoshinga et al., 1995 Japan d < 0.21 -7.2 <0.79 35 ICPMS

Smythe etal., 1982 Australia 3 - 80 (80%) d 2.3 ±0.8 7 XRF

60 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Yoshinga et a!., 1995 Japan 61-96 d 80.5 db 19.6 78.5 45 AAS 17 M, 28 F &ICPAES Kniewald et al., 1994 Croatia f 515.3-592.6 5 ICPAES Zaichick, 1994 d 418±61 NAA,XRF &IGAA Samudralawar and Robertson, 1993 USA 60-82 f 58±17(c/) 55 12 PIXE 8M.4F Robertson etal., 1992 USA 60-82 f 39-97(e) 62 ± 18 (c) 56 12 PKE 38 -107 (r) 60 ± 18 (t) 55 8M.4F

Katie etal., 1991 36-77 d 50 - 72 (te) 67.5 ± 6.8 (te) 174 ICPAES Edward, 1990 65 w 90.1 ± 10.3 1,M NAA Yoshinaga et al., 1989 Japan 0.17-82 d 84.4 ±20.9 83.3 42 ICPAES 28M,14F Kosugi et al, 1988 Japan d 48.1 x/, 1.3 18 ICPAES 11M.7F Lin and Wen, 1988 China >10 a 213.8 ± 109.4 (M) 23, M NAA 251.6 ± 131.4 (F) 24, F Jaksic etal, 1987 66 m 105 (ma) 3 XRF 90 (e) 3 69 (s) 30 94 (ci) 4 108 (cm) 4 65 (ma) 3 PIXE 141(e) 3 258 (ci) 4 174 (cm) 4

61 Age Sample Number of Analytical Reference Location .vs. Preparation Range Mean Median Samples Technique Zwanziger et al, 1987 & 1985 d? 40-112 NAA Mahanti and Barnes, 1983 a 149 ±2 3 ICPAES Knuuttila et a!., 1982 d? 65.5 AAS Gawlik et al, 1982 d 79 ±23 NAA Nieseetal, 1982 d? 48.8 (op) NAA 71.7 (pm) 48.2(0/) Smythe et al, 1982 Australia 3 - 80 (80%) d 62 ±27 7 XRF Hyv3nen-Dabek et al, 1981 24-70 f 26.7-73.0 47.7 ±14.3 15 PKE 9M,6F O'Connor etal, 1980 Australia 20-77 a 72-281 121 ±41 33 XRF 21M.12F

Yoshinga et al, 1995 Japan d < 0.008-0.07 < 0.008 35 ICPMS

Lin and Wen, 1988 China >10 a 0.04 ± 0.020 (M) 15, M NAA 0.047 ± 0.037 (F) 15, F

Yoshinga etal, 1995 Japan d < 0.004-0.02 < 0.004 35 ICPMS

Zaichick, 1994 d 0.0344 ±0.0083 NAA

Lin and Wen, 1988 China >10 a 0.013 ± 0.034 (M) 19, M NAA

62 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique 0.015 ± 0.0056 (F) 13, F

Yoshinga et al., 1995 Japan d <0.3 35 ICPMS

¥»{a

Yoshingaetal., 1995 Japan d <0.06 35 ICPMS

Lin and Wen, 1988 China >10 a 0.38 ± 0.23 (M) 13, M NAA 0.24 ± 0.094 (F) H,F

3t

Yoshingaetal., 1995 Japan 61-96 d <3.0 <3.0 45 AAS 17 M, 28 F & ICPAES Yoshinagaetal., 1989 Japan 0.17-82 d <5.0 0 ICPAES

Kosugi et al., 1988 Japan d 1.95 x/. 1.3 11(7M,4F) ICPAES <1.0 7(4M,3F) Mahanti and Bames, 1983 a 0.5 ±0.01 3 ICPAES

Yoshingaetal., 1995 Japan d < 0.008 35 ICPMS

63 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Yoshinga et al., 1995 Japan d < 0.004 35 ICPMS

tJ{sng%>

Yoshinga et al., 1995 Japan d < 0.004 35 ICPMS

Edward, 1990 65 w <0.5 1,M NAA Lianqing and Guiyun, 1990 China 21 ->61 a 0.061-0.062 30 LF 15M.15F Igarashi et al., 1987 Japan 37-94 w 0.00021-0.00092 0.00067 ±0.00026 6 FT 3M.3F 0.00033-0.00133 0.00083 ±0.00036 7 FT 4M.3F 0.00032 - 0.00065 0.00055 ±0.00019 7(4M,3F) FT Narayan et al., 1987 USA a 0.002-0.40 Wrennetal.,1985 USA a 0.0009-0.023 AAS Fisenne et al., 1984 Russia a 0.011 5 AAS Fisenne et al., 1983 Australia a 0.0035 8 AAS Fisenne et al., 1983 Nepal a 0.016 9 UNSCEAR, 1982 0.022 Fisenne et al., 1980 USA a 0.0008 R ICRP, 1979 0.022

64 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Yoshinga etal., 1995 Japan 61-96 d <6.0 <6.0 45 AAS 17M.28F & ICPAES Yoshinaga et al, 1989 Japan 0.17-82 d <5.0 0 ICPAES

Kosugi etal., 1988 Japan d 4.09 x/, 1.3 11(7 M.4F) ICPAES <2.0 7(4M,3F) Mahanti and Barnes, 1983 a 1.1 ±0.03 3 ICPAES

Yoshinga et al., 1995 Japan d < 0.08-1.2 <0.08 35 ICPMS

Pietra et al, 1993 Italy w 0.0007-0.00275 NAA

Yoshinga et al, 1995 Japan d < 0.03-0.3 <0.03 35 ICPMS

Yoshinga et al, 1995 Japan d < 0.004 -0.02 < 0.004 35 ICPMS

65 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Baranowska et al., 1995 Poland 26-55 f 85.7-138.3 118.22 ± 12.96 25 (15 M, 10 F) AAS 86.4 -138.8 107.41 ± 15.22 10(7M,3F) Bush etal., 1995 18-85 d 71-157 (c) 114± 18.1 (c) 30 ICPAES 30 (18 M, 12 F) Yoshinga et al., 1995 Japan 61-96 d 149 ±19 142 45 AAS 17 M, 28 F & ICPAES Kniewald et a!., 1994 Croatia f 33-88 5 V Pietraetal., 1993 Italy w 40.5-63 NAA Zaichick, 1994 d 91.0 ±4.4 NAA &XRF Samudralawar and Robertson, 1993 USA 60-82 f 144±17(e/) 139 12 PKE 8M.4F Robertson etal., 1992 USA 60-82 f 115-293(c) 182±47(e) 179 12 PKE 117-181(f) 144 ±18(0 139 8M,4F Katie etal., 1991 36-77 d 258-282 (fe) 265.8 ± 15.4 (te) 174 ICPAES Saltzman et al., 1990 USA 20-74 w 42.01±9.11(M) 21 M AAS 46.79 ± 4.70 (F) 5F 54.26 ± 12.28 (M) 21 M 57.87 ± 11.03 (F) 5F 38.62 ± 8.59 (M) 21 M 33.71 ± 4.25 (F) 5F 25.96 ± 6.52 (M) 21 M 29.83 ± 2.79 (F) 5F Yoshinagaetal., 1989 Japan 0.17-82 d 139 ±25 134 42 ICPAES 28M.14F Kosugi et al., 1988 Japan d 120 x/, 1.2 18 ICPAES

66 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique 11M,7F Lin and Wen, 1988 China >10 a 178.8 ± 97.2 (M) 23, M NAA 148.9 ± 82.8 (F) 24, F Jaksic et al., 1987 66 m 187 (ma) 3 XRF 255 (e) 3 221 (s) 30 325 (ci) 4 343 (cm) 4 407 (ma) 3 PKE 353 (e) 3 429 (ci) 4 426 (cm) 4 Valkovicetal., 1987 2-75 m 53 - 855 (s) 10 XRF 195 -1010 (cm) 7M,3F 100 -1060 (ci) Zwanzigeretal., 1987 & 1985 d? 59-244 NAA Jaworowski et al., 1985 France 34-89 d 78 -170 126 ±21 22? ICPES Mahanti and Barnes, 1983 a 102.0 ±1 3 ICPAES Lappalainen et al., 1982 d? 113.9 ±40.7 138 AAS 87M,51F Gawliketal., 1582 d 151 ±22 NAA Sraythe et al., 1982 Australia 3 - 80 (80%) d 98 ±16 7 XRF Hyvonen-Dabeketal., 1981 24-70 f 96.9 -181 144 ±27 15 PKE 9M.6F Lindh, 1981 Sweden 38-186 3 PKE 30-129 3 O'Connor etal., 1980 Australia 20-77 a 174-299 221 ±30 33 XRF 21M,12F

67 Age Sample Number of Analytical Reference Location yrs. Preparation Range Mean Median Samples Technique

Yoshingaetal., 1995 Japan d < 0.3-0.4 <0.3 35 ICPMS

Lin and Wen, 1988 China >10 a 44.30 ± 46.63 (M) 15, M NAA 42.69 ± 29.92 (F) 13, F

68 Table Notations used in Teeth Compilation a = ashed e — enamel d = dry weight basis se = surface enamel f= freeze dried d = dentine m = macerated cd = coronal dentine n = no sample preparation id = inner dentine od - outer dentine sd = surface dentine c = cementum p =pulped cpd = circumpulpal (or secondary) dentine pd = pulpal dentine pf=pulp free

AAS = atomic absorption spectrometry C = colorimetry CPAA = charged particle activation analysis FT = fission track method IGAA = instrumental gamma-activation analysis ICPAES = inductively-coupled plasma atomic emission spectrometry ICPMS = inductively-coupled plasma mass spectrometry ISE = ion-selective electrode MS = mass spectrometry NAA = neutron activation analysis PIXE = proton induced x-ray emission PIGE = proton induced gamma-ray emission PGAA = prompt-gamma activation analysis XRF = x-ray fluorescence V = voltametry

ICS = in-house calibration standard L = no QA/QC mentioned but agreement with the literature values

* not clear f* = Fergusson and Purchase, 1986 (Review article), not clear B* = Bercovitz et al, 1993, not clear.

69 Table 4 A: Major, Minor and Trace Elemeiit in Human Teeth (The references include onlv permanent teeth i.e. no deciduous teeth)

Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique

Johansson, 1991 * 0.08 4 ICPMS Cutress, 1979 14 Countries d 0.2- 396 (se) 32 (se) Z(se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-36.6 (e) 3.44 (e) 244 MS Curzon and Losee, 1978 USA Oregon <20 0.26 ± 0.07 (e) 41 MS California <20 0.06 ± 0.01 (e) 42

Vrbic et al., 1987 Yugoslavia Dalmatia 38-61 n 2.1 ± 1.08 (e) 20 AAS Cutress, 1979 14 Countries d 16- 2304 (se) 343 (se) 202 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0- 510.0 (e) 22.89 (e) 335 MS

Lindh and Tveit, 1980 <14(e) 1M PKE (20 spots)

Ward, 1987 New Zealand 69-84 d 4.12 ±1.13 14 PGAA Cutress, 1979 14 Countries d 0.8 - 13.0 (se) 5.3 (se) 3.6 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-190.0 (e) 8.40 (e) 337 MS Curzon and Losee, 1978 USA Oregon <20 5.79 ± 1.87 (e) 40 MS California <20 0.87 ± 0.16 (e) 42

70 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique

Manea-Krichtenetal., 1991 USA 67-96 d 2.83 -15.7 (e) 6.4 (e) 6 (2M, 4F) MS Vrbicetal., 1987 Yugoslavia Dalmatia 38-61 n 7.7 ± 1.8 (e) 20 AAS Cutress, 1979 14 Countries d 0.8-432 (se) 22 (se) 7(se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-510.0 (e) 18.83 (e) 334 MS Curzon and Losee, 1978 USA Oregon <20 5.13 ± 1.18 (e) 40 MS California <20 2.02 ± 0.69 (e) 42

Cutress, 1979 14 Countries d 0 - 0.04 (se) 0.001 (se) 0(se) 3 MS

Cutress ,1979 14 Countries d 0-6.1(se) 1.3 (se) 1.2 (se) 25 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-15.9 (e) 1.36 (e) 241 MS

M611er and Carlsson, 1984 Sweden Children 1.43±1.29-1.50±1.23(cd) 11 PKE USA New York City 0.57±0.96-1.23±0.59(c

Nowak, 1995 Poland Urban (Industrial) 11-60 d 9.97 ±1.55 62 AAS Rural d 11.7 ±1.6 102 Manea-Krichten et al., 1991 USA 67-96 d 35.2-37.3 (e) 36.5 (e) 6 (2M, 4F) MS Knuuttila et al., 1985 10-74 d 35.5 ± 4.3 (e) 40 AAS

71 Age Sample #of Analytical Reference Location Area vrs Prep Range Mean Median Samples Technique 24M,16F Cohen etal., 1981 Australia d 31.5 ±2.1 (e) 15 PKE 31.5 ±1.1 (d) Lindh, 1981 65 d 37.34 (e) 1M PKE LindhandTveit, 1980 36.6-37.1(e) 36.9 (e) 1M PKE (20 spots)

Nowak, 1995 Poland Urban (Industrial) 11-60 d 2.2 ±0.89 62 AAS Rural d 1.7 ±0.77 102 Cleymaet et al., 1991 Belgium Urban (Industrial) 6-12 * 9.1 ± 8.02 (se) 7.1 249 AAS Kenya Rural 6.4 ±3.8 (re) 5.4 60 Grandjean and Jorgensen, 1990 Greenland Ummannaq-Nuuk 10-43 d 0.042- 0.363 (cpd) 0.086 (cpd) 14 AAS 7M.7F Denmark Funen 10-62 0.035- 0.314 (cpd) 0.097 (cpd) 33 28M.5F Vrbic etal., 1987 Yugoslavia Dalmatia 38-61 n 0.03 ± 0.02 (e) 20 AAS Lappalainen and Knuuttila, 1979 Finland 5 Areas 10-72 d 2.7-6.3 4.2 ±0.7 124 . AAS Cutress, 1979 14 Countries d 0.6- 7.6 (se) 2.7 (se) 1.8 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-27.0 (e) 1.87 (e) 334 MS Curzon and Losee, 1978 USA Oregon <20 1.56 ± 0.32 (e) 40 MS California <20 0.31 ± 0.06 (e) 42 Oehme etal., 1978 Norway Urban Teens d < 0.04-16.2 10 V

Cohen etal., 1981 Australia d 1600- 2800 (e) 15 PKE 2500- 2600 (d) Lindh, 1981 65 d 7900 (e) 1M PKE

72 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Lindh and Tveit, 1980 * 8600 - 9300 (e) 8900 (e) 1M PKE (20 spots)

Cutress, 1979 14 Countries d 0-6.1 (a;) 0.6 (se) 0(se) 23 MS

Nowak, 1995 Poland Urban (Industrial) 11-60 d 6.51 ± 1.75 62 AAS Rural d 4.8 ± 2.42 102 Ngwenya and Turkstra, 1985 South Africa 3 Areas d 0.41 - 2.69 {d) AAS *3 Ethnic groups d 0.12-1.61 (e) Lappalainen and Knuuttila, 1982 Finland 10-76 d 0.0- 0.3 (d) 0.1 ±0.1 (d) 123 AAS 72M,51F Lindh and Tveit, 1980 <16(e) 1M PKE (20 spots) Lappalainen and Knuuttila, 1979 Finland 5 Areas 10-72 d 18.3-34.8 24.8 ±3.0 124 AAS Cutress, 1979 14 Countries d 0-2.7(se) 0.2 (se) 0.1 (se) 47 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0 -1.5 (e) 0.27 (e) 246 MS

Nowak, 1995 Poland Urban (Industrial) 11-60 d 47.2 ± 7.87 62 AAS Rural d 42.6 ± 6.79 102 Ngwenya and Turkstra, 1985 South Africa 3 Areas d 0.83 -3.28(rf) AAS *3 Ethnic groups 1.00- 3.90 (d) COL 0.07-0.45 (e) AAS 0.13-0.62 (e) COL

73 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Cohen etal., 1981 Australia d 20-40(e) 15 PKE 15-30(tf) Coles etal., 1980 England d 0.4 37? AAS Lindh and Tveit, 1980 <34(e) 1M PKE (20 spots) Cutress, 1979 14 Countries d 0.2-4.7(5e) 1.1 (M) 0.7 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-18.0 (e) 0.45 (e) 236 MS

Cutress, 1979 14 Countries d 0 -1.9 (se) 0.1 (se) 0(se) 23 MS

Nowak, 1995 Poland Urban (Industrial) 11-60 d • 9.74 ±30.6 62 AAS Rural d 6.8 ±25.25 102 Vrbic etal., 1987 Yugoslavia Dalmatia 38-61 n 0.92 ± 0.76 (e) 20 AAS Ngwenya and Turkstra, 1985 South Africa 3 Areas d 0.09 -1.20 (d) AAS *3 Ethnic groups 0.64- 2.75 (d) COL 0.14-0.84 (e) AAS 0.07-1.00 (e) COL Moller and Carlsson, 1984 Sweden Children 0.79±1.35 -1.30±1.03 (cd) 11 PKE USA New York City 3.77±13.05 - 5.11±13.30 (cd) 12 Lappalainen and Knuuttila, 1982 Finland 10-76 d 0.5-11.8 (d) 3.0 ± 2.0 (d) 123 AAS 72M.51F Cohen etal., 1981 Australia d 10-30(e) 15 PDflE 15 - 50 (d) Lindh and Tveit, 1980 10 - 26 (e) 21 («) 1M PKE (20 spots)

74 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Lappalainen and Knuuttila, 1979 Finland 5 Areas 10-72 d 7.5-22.7 9.76 ± 2.88 124 AAS Cutress, 1979 14 Countries d 13 -1260 (se) 282 (se) 241 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0 - 30 (e) 1.50 (e) 336 MS Curzon and Losee, 1978 USA Oregon <20 0.21 ± 0.04 (e) 40 MS California <20 0.68 ± 0.15 (e) 41 Oehme et al., 1978 Norway Urban Teens d 1.3 -16.8 10 V

Knuuttila et al., 1985 10-74 d 110±40(e) 40 ISE 24M,16F Cutress, 1979 14 Countries d 25 - 1948 (se) 752 (se) 666 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 8.6 - 925.0 (e) 130.27 (e) 337 MS Curzon and Losee, 1978 USA Oregon <20 54.3 (e) MS California <20 55 (e)

Nowak, 1995 Poland Urban (Industrial) 11-60 d 32.03 ±12.11 62 AAS Rural d 34.4 ±16.01 102 MdllerandCarlsson, 1984 Sweden Children 2.52±2.82 - 3.06±2.36 (cd) 11 PKE USA New York City 6.12±8.66-7.02±8.02(

75 Age Sample # of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique

Cutress, 1979 14 Countries d 0-32(«0 6(se) 5(se) 29 MS

Cutress, 1979 14 Countries d 0.5- 39.6 (se) 7.6 (se) 4(se) 54 MS «£#ftga#

Johansson, 1991 * 0.2 4 ICPMS Lindh and Tveit, 1980 * <22(e) 1M PKE (20 spots)

Cutress, 1979 14 Countries d 0-4.7 (.ye) 0.05 (se) 0.05 (se) 26 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-9.9 (e) 2.04 (e) 92 MS

Nowak, 1995 Poland Urban (Industrial) 11-60 d 53.47 ±13.45 62 AAS Rural d 53.3 ± 8.36 102 Lindh and Tveit, 1980 * 900 -1600 (e) 1200 (e) 1M PKE (20 spots) Curzon and Crocker, 1978 USA & New Zealand 10-20 59.6- 4056.0 (e) 961.41 (e) 337 MS Curzon and Losee, 1978 USA Oregon <20 333.95 ± 42.61 (e) 40 MS California <20 191.04 ± 30.33 (e) 42 La£tt>g&g)

Cutress, 1979 14 Countries d 0-7.2(se) 1.4 (se) 0.8(se) 51 MS

76 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique

Vrbic etal., 1987 Yugoslavia Dalmatia 38-61 n 0.53 ± 0.14 (e) 20 AAS Cutress, 1979 14 Countries d 0.3 -58 (se) 14 (se) 10 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-13.2 (e) 0.92 (e) 246 MS

Knuuttilaetal., 1985 10-74 d 2400 ± 500 (e) 40 AAS 24 M, 16 F Lappalainen and Knuuttila, 1982 Finland 10-76 d 5400-11100 (d) 7926 ± 1044 (d) 123(72M, 51 AAS F) Cohen etal., 1981 Australia d 1000-1100 (e) 15 PKE 3000-8000(0 Lindh, 1981 65 d 4100 (e) 1M PKE Cutress, 1979 14 Countries d 115-3600(re) 745 (se) 576 (se) 54 MS MiU-rog&g)

Nowak, 1995 Poland Urban (Industrial) 11-60 d 47.65 ± 16.93 62 AAS Rural d 39.3 ± 10.2 102 Ngwenya and Turkstra, 1985 South Africa 3 Areas d 0.98- 2.96 (d) AAS *3 Ethnic groups 1.58- 3.00 (d) COL 0.20-1.62 (e) AAS 0.09-2.02 (e) COL Lappalainen and Knuuttila, 1982 Finland 10-76 d 0.4-17.1 (d) 3.9 ±2.5 (d) 123 AAS 72M.51F Cohen etal., 1981 Australia d 5-25<«) 15 PKE 5 - 20 (d) Lindh and Tveit, 1980 <25(e) 1M PKE

77 Age Sample Hot Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique (20 spots) Lappaiainen and Knuuttila, 1979 Finland 5 Areas 10-72 d 5.2-28.9 9.06 ±3.43 124 AAS Cutress, 1979 14 Countries d 2.6-468 (.se) 59(«) 33 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-6.7 (e) 0.60 (e) 336 MS

Jasimetal., 1988 Iraq Baghdad 15-60 d 3.2-8.2 (p/) 5 AAS 6.8-9.2 (p) 5 Vrbicetal., 1987 Yugoslavia Dalmatia 38-61 n 0.84 ± 0.38 (e) 20 AAS Cutress, 1979 14 Countries d 0.04- 0.5 (se) 0.1 (se) 0.04 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0- 32.0 (e) 2.37 (e) 334 MS

Lindh, 1981 65 d 0.14 (e) 1M PKE

Nowak, 1995 Poland Urban (Industrial) 11-60 d 1125.96 ±74.98 62 AAS Rural d 1192.8 ±169.73 102 Cohen et al, 1981 Australia 1700-2600 (e) 15 - PKE 2000- 2300 (d) Lindh, 1981 65 d 13600 (e) 1M PKE

Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-1.0 (e) 0.17(e) 245 MS

Nowak, 1995 Poland Urban (Industrial) 11-60 d 8.42 ±1.35 62 AAS Rural d 4.6 ±1.32 102

78 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Lappalainen and Knuuttila, 1982 Finland 10-76 d 0.4-1.7 (d) 0.9 ± 0.7 (d) 123 AAS 72M,51F Cohen etal., 1981 Australia d 5-15(<2) 15 PKE 5 - 50 (d) Lindh and Tveit, 1980 <13(e) 1M PKE (20 spots) Lappalainen and Knuuttila, 1979 Finland 5 Areas 10-72 d 23.1-44.8 31.3 ±4.4 124 AAS Cutress, 1979 14 Countries d 0.4- 270 (se) 23 (se) 9(se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-9.0 (e) 0.90 (e) 249 MS Curzon and Losee, 1978 USA Oregon <20 0.29 ± 0.09 (e) 38 MS California <20 1.16 ± 0.19 (e) 40

Knuuttila et al., 1985 10-74 d 16.5 ± 1.4 (e) 40 C 24 M, 16 F Cohen etal., 1981 Australia d 16.1 ±1.5 (e) 15 PDCE 14.0 ±1.8 (d) Lindh, 1981 65 d 17.38 (e) 1M PKE Lindh and Tveit, 1980 19.9-21.5 (e) 21 («) 1M PKE (20 spots)

Nowak, 1995 Poland Urban (Industrial) 11-60 d 47.65 ± 16.93 62 AAS Rural d 39.3 ± 10.2 102 Gil etal., 1994 Spain Urban & Rural 20->60 d 0.13-80.37 6.15 x/- 1.21 40 AAS Bercovitz et al., 1993 Israel 5 Regions 14-75 d 0.58-36.89 1.65-25.72 180 AAS Bercovitz and Laufer, 1993 Israel Northern Region d 1.24-32.72 (d) 10.95 ±9.11 id) 22 AAS

79 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Bercovitz and Laufer, 1991 Israel Northern Region 31-75 d 2.11 - 117.37(rf) 25.62± 10.15(41 12 AAS Cleymaetetal., 1991 Belgium Urban (Industrial) 6-12 * 1236 ± 849 (se) 1066 252 AAS Kenya Rural 145±67(se) 134 60 Manea-Krichtenetal., 1991 USA 67-96 d 4.1-29.8 (e) 14 (e) 6 (2M, 4F) MS Frank etal., 1990 Mexico Urban 12-29 d 24.1 ± 9.3 (od) 118.8 ± 95.6 (id) France Strasbourg 32-65 d 45.3 ± 15.8 (id) Grandjean and Jorgensen, 1990 Greenland Ummannaq-Nuuk 10-43 d 5.0 -172 (cpd) 16.8 (cpd) 14 AAS 7M,7F Denmark Funen 10-62 3.5 -163 {cpd) 23.7 (cpd) 33 28M.5F Khadekar et al., 1986 India Bombay ? 2.0-28.0 8.31x/, 1.20 71 V 30, M, 41 F Purchase and Fergusson 1986 New Zealand Urban d 99 (sd) 5 AAS 20.4 (d) 8 7.9 (e) 8 1100(«!) 13 14.8 (d) 171 14.3- 29.6 (e) 17 Grobleretal., 1985 South Africa Urban & Rural 6-12 d 26.5-74.5 Kollmeier etal., 1984 Germany Urban 10-70 d 1-30 163 AAS Ogawa, 1983 Japan d 1.55(0 AAS 1.7 (e) Lappalainen and Knuuttila, 1982 Finland 10-76 d 1.0-39.0(rf) 13.7 ±8.5(0 123 AAS 72M,51F Cohen etal., 1981 Australia d < 10 (e) 15 PKE 10-30(rf) Steenhout and Pourtois, 1981 Belgium Industrial a 5.4-110.7 33.35 51 AAS

80 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Belgium Rural 1.2-69.3 21.45 38 Belgium Urban 2.3-68.0 17.72 42 Al-Naimietal., 1980 England Birmingham 12-16 d 17.3- 43.9 (pd) 34.2 ± 17.0 (pd) 10 CPAA 40-71 21.0-213.1 (pd) 92.2 ±31.9 (pd) 19 12-16 5.8 -12.6 (d) 7.2 ± 3.4 (d) 10 40-71 5.0- 39.6 (d) 17.3 ± 7.3 (d) 19 12-16 1.6-3.6 (e) 2.3 ± 1.0 (e) 10 40-71 1.4-5.3 (e) 2.7 ± 0.7 (e) 19 Sheffield 12-16 12.5-24.5 (pd) 18.9 ± 7.4 (pd) 5 40-72 28.2 - 299.6 (pd) 112.5 ± 52.9 (pd) 11 12-16 3.8-10.4 (d) 7.2 ± 2.9 (d) 5 40-72 5.5 - 66.7 (d) 25.6 ± 12.4 (d) 11 12-16 1.4-3.5 (e) 2.1 ± 1.0 (e) 5 40-72 1.2-5.4 (e) 4.0 ± 0.7 (e) 11 Aberystwyth 5-15 20.8-54.3 (pd) 27.5 ± 9.6 (pd) 10 40-70 112.6-310.3 (pd) 149.3 ± 92.5 (pd) 5 5-15 4.2-11.1 (d) 5.6 ± 2.0 (d) 10 40-70 26.9 - 81.0 (d) 40.6 ± 22.8 (d) 5 5-15 1.6-2.9 (e) 2.0 ± 0.6 (e) 10 Coles et al., 1980 England d 2.0 - 5.0 3.8 37 AAS Lindh and Tveit, 1980 <21(e) 1M PKE (20 spots) Cutress, 1979 14 Countries d 1.2-79(«0 24 (ye) 18 (se) 54 MS Grandjeanetal., 1979 Denmark Copenhagen 16-54 n 25.7 (cpd) 17 V 9M.8F Lappalainen and Knuuttila, 1979 Finland 5 Areas 10-72 d 30-79.2 53.5 ± 7.8 124 AAS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0 -156.0 (e) 19.64 (e) 335 MS Oehmeetal., 1978 Norway Urban Teens d 1.8-4.9 10 V

81 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Pinchinetal., 1978 England Urban d 1.22-3.60 8 V Khandekaretal., 1978 India Urban d 4.27-82.5 72 AAS Attramadal and Jonsen, 1976 Norway Urban Teens d 0.9-7.8 44 V Wilkinson and Palmer, 1975 USA Mixed 0-79 d H-55 336 AAS Rural 0-79 d 14-67 230 Urban 6-59 d 0-38 62 Shapiro et al., 1975 USA Non-industrial d 1-30 163 AAS Kaneko et al., 1974 Japan 4 Areas d 1.12-64.67 93 AAS Malik and Fremlin, 1974 England Urban 19-57 d 25 - 52.7 10 CPAA Stack etal, 1974 Scotland Rural Teens d 17-20 36 AAS Langmyhr et al., 1974 Sweden d 1.1-6.4 14 AAS Strehlow and Kneip, 1969 USA 62 a 254-336 4 AAS < 10-60 a 17-16

Cutress, 1979 14 Countries d 0 - 4.7 (se) 0.2 (se) 0(se) 14 MS

Cutress, 1979 14 Countries d 0.1 -4.0 (se) 0.6 (se) 0.4 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0- 30.0 (e) 4.61 (e) 256 MS Curzon and Losee, 1978 USA Oregon <20 0.33 ± 0.07 (e) 32 MS California <20 0.15 ± 0.02 (e) 33

Cohen etal., 1981 Australia d 1000 -1350 (

82 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Lindh and Tveit, 1980 * 812-819 (e) 815 (e) 1M PKE (20 spots) Cutress, 1979 14 Countries d 1836- 40320 (se) 18780 (se) 17280 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0 - 200.0 (e) 24.19 (e) 336 MS

Cutress, 1979 14 Countries d 0-90(«!) 8(«) 3(se) 26 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-3.0 (e) 0.20 (e) 225 MS Curzon and Losee, 1978 USA Oregon <20 0.07 ± 0.0 l(e) 36 MS California <20 0.02 ± 0.00 (e) 41

Ngwenya and Turkstra, 1985 South Africa 3 Areas d 0.27 -1.03 (d) F- *3 Ethnic groups d 0.06-1.03 (e) Cutress, 1979 14 Countries d 2.9-72(se) 18 (se) 16 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0 -18.1 (e) 1.47 (e) 326 MS

Cutress, 1979 14 Countries d 1.3- 504 (se) 70 (se) 40 (se) 54 MS Sn(tag/feg)

Lindh and Tveit, 1980 <325(e) 1M PKE (20 spots) Cutress, 1979 14 Countries d 0.9-72(«0 9.3 (se) 5.8 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0- 44.0 (e) 1.60 (e) 245 MS

83 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Frank etal., 1989 France Strasbourg 10-27 n 150.5 ± 55.6 (se) 8 XRF 155.0 ± 51.4 (ie) 94.6 ±43.6 (peed) 92.6 ± 35.6 (med) 87.2 ± 34.7 (ped) 82.9 ± 32.0 (perd) 77.7 ± 30.1 (prd) 32-65 196.9 ± 62.3 (se) 7 206.4 ± 76.2 (ie) 136.7 ± 60.2 (peed) 124.3 ± 41.6 (med) 125.0 ±37.6 (pa/) 123.2 ±46.9 (perd) 132.8 ± 57.3 (prd) Vrbicetal., 1987 Yugoslavia Dalmatia 38-61 n 106.7 ± 23.1 (e) 20 AAS Mdller and Carlsson, 1984 Sweden Children 32.2- 205.0 (cd) 83.3 ± 52.1 (cd) 11 PKE USA New York City 23.5-251.0 (erf) 75.0 ± 59.9 (cd) 12 Curzon and Cutress,1983 70-286 (e) 115 (e) Curzon.1983 150 (d) Lappalainen and Knuuttila, 1982 Finland 10-76 d 32.5 -138.0 (d) 80.7 ± 24.3 (d) 123(72M, 51 AAS F) Chaudhari and Crawford, 1981 23 Cohen etal., 1981 Australia d 100-150 (e) 15 PKE 110-150 0) Lindh and Tveit, 1980 * 96 -108 (e) 103 (e) 1 M (20 spots) PKE Cutress, 1979 14 Countries d 9-7632(se) 204 (se) 36 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 13.0 -1400.0 (e) 157.15 (e) 337 MS Curzon and Losee, 1978 USA Oregon <20 81.51 ± 9.04 (e) 33 MS

84 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique California <20 130.51 ± 12.27 (e) 41

Cutress, 1979 14 Countries d 0.1- 24.5 (se) 1.6 (se) 0.6 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-31.4(e) 1.93 (e) 243 MS

Vrbicetal., 1987 Yugoslavia Dalmatia 38-61 n 0.0039 ± 0.0022 (e) 20 NAA Cohen etal., 1981 Australia d 10-30(e) 15 PKE 15-35(<0 Byrne and Vrbic, 1979 Yugoslavia Zemunik 0.0038 ± 0.0013 (e) 9 NAA Novigrad 0.0041 ± 0.0022 (e) 8 Ljubljana 0.0031 ± 0.0012 (e) 10 Belgrade 0.0033 ± 0.00 ll(e) 5 Cutress, 1979 14 Countries d 0.1- 14.4 (se) 1.4 (se) 0.5 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0-0.2 (e) 0.02 (e) 239 MS

Cutress, 1979 14 Countries d 0-9.3 (re) 1.8 (re) 0.9 (se) 29 MS

Lindh and Tveit, 1980 * <325(e) 1M PDfE (20 spots)

Nowak, 1995 Poland Urban (Industrial) 11-60 d 357.08 ± 267.2 62 AAS Rural d 228.3 ± 160.84 102 Knuuttila etal., 1985 10-74 d 240 ± 100 (e) 40 AAS 24 M, 16 F

85 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique Frank etal., 1989 France Strasbourg 10-27 n 228.1 ± 152.7 (se) 8 XRF 69.7 ± 19.5 (ie) 147.3 ± 27.3 (peed) 169.3 ± 26.6 (med) 234.5 ± 54.2 (ped) 142.0 ±31.1 (perd) 263.8 ± 67.7 (prd) 32-65 261.6 ±188.6 (ie) 7 115.9 ± 94.4 (ie) 158.2 ± 28.8 (peed) 190.9 ± 38.7 (med) 325.9 ± 107.6 (ped) 153.4 ±24.9 (perd) 359.3 ±161.4 (prd) Ngwenya and Turkstra, 1985 South Africa 3 Areas d 152-203 (d) AAS *3 Ethnic groups d 133 - 180 (e) Moller and Carlsson, 1984 Sweden Children 102.0- 165.0 (cd) 135.1 ± 21.2 (cd) 11 PKE USA New York City 83.0 - 533.0 (cd) 163.0 ± 120.0 (cd) 12 Curzon and Cutress, 1983 126-276 (e) 210 (e) Lappalainen and Knuuttila, 1982 Finland 10-76 d 50.0- 999.0 (d) 157.0 ± 87.9 (d) 123 AAS 72M.51F Chaudhari and Crawford, 1981 Australia d 100-300 (e) 15 PKE Cohen etal., 1981 103 200- 900 (d) Coles etal., 1980 England d 141 37? AAS Lindh and Tveit, 1980 * 676-702 (e) 690 (e) 1M PKE (20 spots) Lappalainen and Knuuttila, 1979 Finland 5 Areas 10-72 d 133 - 5600 182 ±37.1 124 AAS

86 Age Sample #of Analytical Reference Location Area yrs Prep Range Mean Median Samples Technique

Cutress, 1979 14 Countries d 61 - 5400 (se) 893 (se) 576 (se) 54 MS Curzon and Crocker, 1978 USA & New Zealand 10-20 9.9 - 806.0 (e) 153.12 (e) 336 MS Oehmeetal., 1978 Norway Urban Teens d 76-542 10 V

Curzon and Crocker, 1978 USA & New Zealand 10-20 0.0- 0.8 (e) 0.08 (e) 244 MS

87 Frontal Parietal Temporal -Zygoma tic Maxilla Mandible 7th cervical vertebra Shoulder f Clavicle ..-- 1st thoracic vertebra Girdle \scapula 1st nb

Sternum Humerus

. .. 12th rib

Forearm Ilium | Pubis 1 Os coxae Ischiumj

Metatarsus •v{\ Phalanges

Figure 1: Bones of the Human Skeleton (Ref. Dorland's Medical Dictionary, W.B. Saunders Co., Philadelphia, PA, 1965). Dentine

Figure 2: Component tissues of a tooth (Ref. Biochemist's Handbook, Long C. (Ed), Van Norstrand Co, New York, 1961). Vertical and transverse cuts with stainless steel Sagittal Saw (3M Co. Ltd.). Rinsed with ultra-pure water; frozen until dissection.

CUT In Class-100Cleanroom: Non-osseous material removed, biopsy cut into 5 or more slices.

Water-jet, dental probes, compressed gas to remove red/white marrow.

Cortical bone trimmed by osteotome/scalpel blade

icm Ultra-pure / water wash, then shake with glass-distilled acetone (1 min) and ultra-pure water (1 min), in Teflon vials. Dry with compressed N2-

Random division of 2 bone types, drived in vacuum desiccator in Teflon vials.

ISOTOPIC RATIO TOTAL LEAD Sealed and transported to analytical ANALYSIS (TIHS) ANA1YSI8(GfAAfl) laboratory. TOTAL LEAO(IDKS)

Figure 3: Dissection and preparation of cortical and trabecular bone sample from a iliac crest bone biopsy {Ref. Inskip et al. Neurotoxicology 13(1992)825}.

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