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— Ute Tana Bib Takin THE EFFECTS OF BORON SUPPLEMENTATION ON BONE MINERAL DENSITY, BLOOD AND URINARY CALCIUM, MAGNESIUM, PHOSPHOROUS AND URINARY BORON IN FEMALE ATHLETES by Susan Meacham Darnton Dissertation submitted to the Faculty of the Virginia Polytechnic Institute and State University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in Human Nutrition and Foods APPROVED: L. Jahette Taper, ps Ute— tana bib takin ' AMI slo . Shoes igf- — W. Pandan U.)Thomas J. W. Rankin September 20, 1991 Blacksburg, Virginia ” Ses BOT \nRes, i i “+: IGG/ L655 VSG D8 LD THE EFFECTS OF BORON SUPPLEMENTATION ON BONE MINERAL DENSITY, BLOOD AND URINARY CALCIUM, MAGNESIUM, PHOSPHOROUS AND URINARY BORON IN FEMALE ATHLETES by Susan Meacham Darnton Committee Chairman: L. Janette Taper Human Nutrition and Foods (ABSTRACT) The effects of boron supplementation on blood and urinary minerals were studied in 17 female college athletes and 11 sedentary college contro! subjects. The subjects were similar in age (19.8 + 1.4 and 20.3 + 1.1 years for athletes and sedentary groups, respectively) and weight (61.8 + 9.1 and 59.6 + 10.5 kgs for athletes and sedentary groups, respectively). The athletic subjects had lower percent body fat averages (20.6 + 5.6 and 25.8 + 6.5 %, respectively, (9<0.05)) and higher aerobic capacities (2.9 + 0.5 and 2.1 + 0.4L 0,*min,, respectively, (P<0.05) than sedentary controls. No differences in dietary intake were observed. Serum phosphorous levels were lower in boron supplemented subjects than in subjects receiving placebos (p<0.05) and were lower during final analysis than during baseline analysis. A group-supplement interaction was noted with serum phosphorous also (p<0.05). In the sedentary group, boron supplementation lowered serum phosphorous and placebo supplementation elevated serum phosphorous. In the athletic group, no changes in serum phosphorous were observed due to supplementation. Serum magnesium was greatest in the sedentary controls supplemented with boron and increased with time in all subjects (p<0.05). Again, a group-supplement interaction was observed with serum magnesium; exercise in boron supplemented subjects lowered serum magnesium (p<0.05). In all subjects, calcium excretion increased over time (p<0.05) and in boron supplemented subjects boron excretion increased over time (p<0.05). In conclusion, boron supplementation affected serum phosphorous and magnesium and the excretion of boron in the urine. The significance of these findings in relation to overall mineral status, bone mineral density, and exercise jin college female athletes needs further investigation. It is possible that increasing the intake of foods high in boron may be found to be beneficial in the treatment and/or prevention of metabolic bone disorders such as those related to excessive exercise. ACKNOWLEDGEMENTS | would like to express my appreciation to my major professor, Dr. L. Janette Taper, for her advice, encouragement and patient teaching throughout this project. | am also very pleased to have had a very special person, Dr. Stella Volpe-Snyder, as a research partner and friend has we progressed through the study. | would also like to thank those individuals on campus and at Montgomery Regional Hospital who have provided their time and facilities for this research effort. | would particularly like to thank Drs. Barbara Chrisley and Kathy Reynolds for their technical assistance and support. In addition, | would like to thank the subjects, faculty, friends and family members, especially my mother, for their time, support and belief in education. iv TABLE OF CONTENTS Page ACKNOWLEDGEMENTS ......... 0.0.0. eens iV LIST OF TABLES .. 0... ne viii , CHAPTER! INTRODUCTION ..........0.0 cee ens 1 DefinitionS 2... ee ees 3 CHAPTER Il REVIEW OF LITERATURE .............0.0000 eens 5 OStEOPOrOSiS ©... ee ee ee ees 5 In elderly femaleS .. 0.0... 0... eee eee 5 In young females... 1... 20... eee 6 Bone mineral density in young female athietes ................. 8 Major Mineral Metabolism in Bone... 0.2... 0.0.00. ee ees 14 Calcium 6 ee ens 15 Phosphorous .... 0.0... ee ees 17 Magnesium 1... 0... ees 20 Boron--An Ultratrace Mineral .... 0.0.0.0...0. cece eee 24 Boron--a general overview ..........0000 cee eee eee 26 Boron in plant metabolism ... 0.0.0.0...00 cee ees 28 Boron-calcium interaction in plants ...... 0.0.0.0...ee ees 30 Boron--health benefits .. 0... 0.0... ees 31 Boron toxicity and boron deficiency ...............000 eee 32 Boron in the diet... 0... eens 35 Is boron an essential nutrient? ......... 0.0.0.0... ees 37 Boron’s physiological role 0... 1.0... eee 38 Boron studies at Grand Forks, North Dakota .................. 40 Boron concentrations in foods and tissues ..............-.005 46 Methodology Review ........ 0.0: cee eee ee eens 48 Mineral analysis using atomic absorptiometry ................. 50 Inductively coupled argon plasma spectroscopy ............... 50 Analysis of boron ow. eee 52 Sample preparation for boron determinations ................. 54 Microwave digestionS ....... 0... 0c ee eee 54 Nitric acid digestions ... 0... eee 55 Wet-ash, low-temperature Teflon tube digestions ............... 56 CHAPTER Ill METHODOLOGY ....... 2... cee eee 57 Subjects and Study Design .........0.0.00. ee ees 57 Subjects .. 1. ee eee 57 Experimental design ...... 0... 0. ccc es 58 Analytical Procedures ... 0... 0... ees 59 Aerobic work capacity .... 0... 0.0.0 ee eee 59 Percent body fat ...... ee ee ees 63 Height and weight ...... 0... 2... 0. ee eee 64 Bone mineral density ........ 0.0.0.0 c ec ee ees 64 Food collection 2... 0... ee ee eee 64 Diet analySiS 2... ne 65 Blood collection ....... 0.0.0... ceee ee eae 65 Blood analysis... 0... eee nee 66 Urine collection 2.0... 0. eens 67 Urine analySiS 2.0... ne 67 Statistical analysis 2.0... ees 68 CHAPTER IV RESULTS ..... ee ene 69 CHAPTER V DISCUSSION ....... 0.0.0.0. ceceee eee 84 Physical CharacteristicS ©... 0.0.0.0... cc ee eee 84 Menstrual status ... 0... ees 84 Method of determining menstrual status ..................04.% 86 Bone mineral density .........0.0.0.0. cee eee eae 87 Dietary Intake Analysis .......0.0.0 0.000000 eeeee 88 Methods of dietary intake analysis ..............000cee eee 92 Baseline and final dietary analysis ............0.0.00 eee 94 Blood Mineral Analysis ....... 0.0.0.0. cee eees 95 Methods of blood calcium analysis ......... 0.0.00. eee eee 97 Urine Mineral Analysis .. 0.0.0... 0.0.00. ee eee ees 98 Methods of urine mineral analysis .............0.0.00 eee ees 100 CHAPTER VI RECOMMENDATIONS FOR FUTURE RESEARCH ....... 102 LITERATURE CITED ...... 0... ccc tenes 103 JOURNAL ARTICLE... 1. eee ens 116 vi APPENDIX A: INSTITUTIONAL REVIEW BOARD APPENDIX B: WRITTEN INFORMATION AND INFORMED CONSENT .. 153 APPENDIX C: PRESTUDY QUESTIONNAIRE .................-0- 157 APPENDIX D: MENSTRUAL RECORDS ................-0.+0004 170 APPENDIX E: POSTSTUDY QUESTIONNAIRE ................... 173 APPENDIX F: INDIVIDUAL DATA ....... 2... 0. eee ee 175 APPENDIX G: UNDERGRADUATE RESEARCH CREDITS FORM ...... 179 APPENDIX H: SUMMARY OF COMPLETE STUDY ANALYSES ....... 182 APPENDIX I: PWC,., AND ANTHROPOMETRIC DATA SHEETS ...... 185 APPENDIX J: BORG’S RATING OF PERCEIVED EXERTION ......... 192 APPENDIX K: SAMPLE DATA SHEET FOR DUAL PHOTON ABSORPTIOMETRY ...............0-005 194 APPENDIX L: FOOD COLLECTION PROCEDURES ............... 196 APPENDIX M: DIETARY FAT ANALYSIS ................ 002.0005. 198 APPENDIX N: ATOMIC ABSORPTION PROCEDURES ............. 201 APPENDIX O: PHOSPHOROUS ANALYSIS .................0004 205 APPENDIX P: NUTRITIONIST Ill PROCEDURES .................. 212 APPENDIX Q: URINE COLLECTION PROCEDURES ............... 215 APPENDIX R: WET ASH DIGESTION PROCEDURES .............. 217 APPENDIX S: ICP USER INFORMATION .............-....-000005 222 APPENDIX T: BLOOD COLLECTION PROCEDURES .............. 230 APPENDIX U: HUMAN WASTE DISPOSAL PROCEDURES .......... 236 APPENDIX V: IONIZED CALCIUM ANALYSIS .................0.. 244 APPENDIX W: RAW DATA TABLES ............00020: eee eee 248 Ce Vil LIST OF TABLES Page Reference concentrations of calcium, phosphorous, magnesium and boron in the diet, blood and urine ........... Boron concentration in various foods and human health care products ...... 0.0.0.0... ccc eens Boron concentrations in various tissueS ..............0000. Assignment of subjects to treatment groups ................ Summary of the experimental design ...............00000s Summary of analytical methods ...................20005- Subjects’ age, body weight, body fat, and VO?max ........... Subjects’ bone mineral density measurements .............. Subjects’ daily dietary intakes assessed by laboratory analyses .. 0.0... ees 10. Comparison of laboratory analyses and Nutritionist III assessments of subjects’ baseline daily dietary intakes ....... 11. Baseline and final comparison of subjects’ daily dietary energy, protein, fat, carbohydrate and minerals assessed by Nutritionist I]... eee 12. Baseline and final comparison of subjects’ blood calcium, phosphorous, and magnesium assessed by atomic absorption 13. Baseline and final comparison of subjects’ plasma calcium assessed by the NOVA 7 ........0.0... eee eee viii 14. Comparison of subjects’ blood calcium assessed by the NOVA 7 and AA... ee nee 80 15. Baseline and final comparison of subjects’ daily urinary calcium, phosphorous, magnesium and boron........... 81 16. Comparison of subjects’ daily urinary calcium, phosphorous and magnesium assessed using AA, SD and ICP .. 1... eens 83 1x CHAPTER | INTRODUCTION
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