Mass spectroscopy techniques for detecting vitamin D compounds in a sample
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1. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
- a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy;
b) determining an amount of 25-hydroxyvitamin D2 in a sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode; and
c) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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Abstract
Methods for determining the amount of vitamin D compounds in a sample are provided. The methods can employ LC-MS/MS techniques and optionally the use of deuterated internal standards. Methods for diagnosing vitamin D deficiencies are also provided.
12 Citations
40 Claims
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1. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) determining an amount of 25-hydroxyvitamin D2 in a sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode; and c) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) determining an amount of 25-hydroxyvitamin D2 in a sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode, and wherein the LC-MS/MS technique comprises a Q1 scan tuned to select a precursor ion that corresponds to the [M+H+] of 25-hydroxyvitamin D2; and c) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
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18. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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d) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; e) determining an amount of 25-hydroxyvitamin D2 in a sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode, and wherein the LC-MS/MS technique comprises monitoring an MRM precursor-product ion pair transition having m/z values of 413.0/395.3 for 25-hydroxyvitamin D2; and f) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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19. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) deriving a sample from the mammalian patient'"'"'s biological sample comprising de-proteinizing the sample; c) determining an amount of 25-hydroxyvitamin D2 in a sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode; and d) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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20. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) deriving a sample from the mammalian patient'"'"'s biological sample by removing interfering components from the sample; c) determining an amount of 25-hydroxyvitamin D2 in the sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode; and d) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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21. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) deriving a sample from the mammalian patient'"'"'s biological sample by removing interfering components from the sample; c) determining an amount of 25-hydroxyvitamin D2 in the sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode, and wherein the LC-MS/MS technique comprises a Q1 scan tuned to select a precursor ion that corresponds to the [M+H+] of 25-hydroxyvitamin D2; and d) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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22. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) deriving a sample from the mammalian patient'"'"'s biological sample by removing interfering components from the sample; c) determining an amount of 25-hydroxyvitamin D2 in the sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive ion mode, and wherein the LC-MS/MS technique comprises monitoring an MRM precursor-product ion pair transition having m/z values of 413.0/395.3 for 25-hydroxyvitamin D2; and d) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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23. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) deriving a sample from the mammalian patient'"'"'s biological sample by removing interfering components from the sample; c) determining an amount of 25-hydroxyvitamin D2 simultaneously with an amount of 25-hydroxyvitamin D3 in a single sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2 and underivatized 25-hydroxyvitamin D3 in the single sample, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive-ion mode; and d) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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24. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) deriving a sample from the mammalian patient'"'"'s biological sample by removing interfering components from the sample; c) determining an amount of 25-hydroxyvitamin D2 simultaneously with an amount of 25-hydroxyvitamin D3 in a single sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2 and underivatized 25-hydroxyvitamin D3 in the single sample, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive-ion mode, and wherein the LC-MS/MS technique comprises a Q1 scan tuned to select, independently, precursor ions that correspond to the [M+H+] of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3; and d) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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25. A mass spectroscopy method for monitoring a vitamin D replacement therapy in a mammalian patient comprising:
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a) obtaining a biological sample from the mammalian patient, wherein the mammalian patient has received at least one dose of a vitamin D replacement therapy; b) deriving a sample from the mammalian patient'"'"'s biological sample by removing interfering components from the sample; c) determining an amount of 25-hydroxyvitamin D2 simultaneously with an amount of 25-hydroxyvitamin D3 in a single sample derived from the mammalian patient'"'"'s biological sample, wherein the determining comprises using a mass spectrometry technique to detect underivatized 25-hydroxyvitamin D2 and underivatized 25-hydroxyvitamin D3 in the single sample, wherein the mass spectrometry technique comprises a LC-MS/MS technique and wherein the LC-MS/MS technique comprises the use of a triple quadrupole instrument in Multiple Reaction Monitoring (MRM), positive-ion mode, and wherein the LC-MS/MS technique comprises monitoring MRM precursor-product ion pair transitions having m/z values of 401.4/383.3 for 25-hydroxyvitamin D3 and 413.0/395.3 for 25-hydroxyvitamin D2; and d) comparing the amount of 25-hydroxyvitamin D2 in the sample with a range for 25-hydroxyvitamin D2 expected for the replacement therapy, wherein a lower than expected amount of 25-hydroxyvitamin D2 relative to the range expected is indicative of patient non-compliance with the therapy, malabsorption of vitamin D supplements, or resistance to 25-hydroxyvitamin D2.
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Specification