. 27 : 237-245 2009 Ann Rept. BFDA Taiwan R.O.C. 27 : 237-245 2009 237 D 97 (online SPE LC-UV) (online SPE LC\MS\MS)D(flame atomic absorption spectrophotometer, FLAAS)32 D 80 1206 8022 1204 35 80 1209 8018 1208 D D 95 D 2 (ergocalciferol) D 3 (cholecalciferol)(hplc) (flame atomic absorption spectrophotometer) D (rickets)d (1) D cholecalciferol D (ergocalciferol, vitamin D 2 ) (cholecalciferol, vitamin D 3 ) (2) (3) D (2) D(mineralization) (demineralization) (rickets) (osteomalacia) 7 mg/ dl (3) D (international units, IU)(μg) D 1 IU0.025 μg cholecalciferol 1 μg cholecalciferol40 IU (2) 25-hydroxyvitamin D (25(OH)D) 400~1,250 nmol/l (160~500 ng/ml) DD (hypercalcemia of hypervitaminmosis D) D
238 Ann. Rept. BFDA 去 氫 膽 固 醇 (7-dehydrocholesterol) 維 生 素 D3(cholecalciferol) 7-dehydrocholesterol 去 氫 膽 固 醇 (7-dehydrocholesterol) D3 cholecalciferol 維 生 素 D3(cholecalciferol) ergosterol 麥 角 醇 (ergosterol) D D2 ergocalciferol 維 生 素 D2(ergocalciferol) (resorption) μg 1,250~5,000 μg (50,000~200,000 IU) 99% 麥 角 醇 (ergosterol) (2) 25 μg (1,000 IU)1~18 50 μg (2,000 IU) 1850 μg 圖 一 維 生 素 D 之 結 構 維 1% 生 素 D2(ergocalciferol) (5) 9~12 mg/dl (2) D (2) (Tolerable Upper Intake Levels) 12 (parathyroid 圖 一 維 生 素 D 之 結 構 hormone, PTH)PTH 1,25- dihydroxyvitamin D (1,25(OH) 2 D 3 ) (2,000 IU)( 14~50 ) Ca http://ceiba3.cc.ntu.edu.tw/ ++ 50 μg (2,000 IU) course/4f786b/infof.htm (3) (2,5) D (CaCO 3 ) (4) 1. (CaSO 4 2H 2 O)(Ca 3 PO 4 2 ) D (CaHPO http://ceiba3.cc.ntu.edu.tw/ 4 )(calcium citrate) 800 IU (20 μg ) 2. course/4f786b/infof.htm (3) ()300 g( (40%) )D15 24
D 239 D D () (μg) 0 10 3 10 6 10 9 10 1 ~ 5 4 ~ 5 7 ~ 5 10 ~ 5 13 ~ 5 16 ~ 5 19 ~ 5 31 ~ 5 51 ~ 10 71 ~ 10 +5 +5 +5 +5 1,500~2,400 mg (37.5 to 60 mmol) (2) () (mg) 0 200 6 300 9 400 1 400 4 500 7 500 10 600 13 800 16 1000 19 800 31 800 51 800 71 800 +0 +0 +0 +0 () (4) 1. 300 g( )1800 mg 2. () 300 g( )750 mg 95D () (%) () 1000 Calcium carbonate 625 40 250 4 Calcium citrate 950 21 200 5 Calcium gluconate 500 9 45 22 Calcium lactate 325 13 42 24 Calcium phosphate, dibasic 500 22.3 115 9 Calcium phosphate, tribasic 800 38 304 4
240 Ann. Rept. BFDA (6) 526 D 80~12035 6 95 6 95D 95 D D ( )96 on-line SPE LC/MS/MS SPE D35 ㈠D 1. Ergocalciferol(D 2 ) cholecalcife r o l ( D 3 ) T h e D 4 2 1 7 4 7 0 11 5 12 0 17 13 21 1 35 United States Pharmacopeial Convention, Inc., Dimethyl sulfoxide (DMSO) methanol Merck, n-hexane acetyl acetate acetonitrile (AcCN)HPLCLabscan Co., Ltd., 2. (Syringe filters) Nylon, 0.45 μm, 13 mm, Titan Inc. UltiMate 3000 Dionex Corporation Ltd. Nucleodur Sphinx 5 μm4.6 250 mm Macherey- Nagel ODS-3V 5 μm 4.6 250 mm GL Sciences : UltiMate 3000 Applied Biosystems API3200 3. 970971800323 D (7) 96 D (8) D 2 D 3 10 mg100 ml n-hexane n-hexane 0.5 1.0 2.0 5.0 10.0 μg/ml 5~10 g DMSO 10 ml40~50 C 30 50% methanol 10 mln-hexane 20 ml 30 3500 rpm 10 0.45 μm
D 241 (online SPE LC-UV) (online SPE LC\MS\MS)D A. Online SPE-LC LCSPE t M ( t M : sample matrix elutçn time from SPE column) B. C. SPE Nucleodur Sphinx 5 μm 4.6 250 mm Macherey- Nagel ODS-3V 5 μm4.6 250 mm GL Sciences D.265 nm Ionization mode: APCI + (9) (10) Scan type: MRM Curtain gas: 10 psi Collisçn gas: 5 psi Source Temperature: 300 C Nebulizer gas: 60 psi Turbo gas: 50 psi Interface heater: on Declustering potential: 50 V Needle current: 4000 kv Entrance Potential: 10 V Collisçn cell exit potential: 4 V Dwell time: 200 msec : Compound Precursor çn (m/z) Product çn (m/z) Collisçn energy (V) Vit D 3 385 367 20 385 259 25 Vit D 2 397 271 37 397 379 37 20 μl UV D Retentçn Time (min) Retentçn Time (min) Flow ml/min Left pump A (%) Retentçn Time (min) Flow ml/min Right pump A (%) B (%) 0 1-2 0 1.0 100 0 1.0 100 0 6.3 6-1 1.0 100 20 1.0 100 0 13.0 1-2 1.0 100 20.1 1.0 98 2 1.0 100 50.0 1.0 98 2 1.0 100 50.1 1.8 100 0 60.0 1.0 100 60.0 1.8 100 0 A: Acetonitr B: H 2 O 1-2SPE 6-1SPE
242 Ann. Rept. BFDA 20 μl MRM (transition ion)(ion ratio) D C x V D(μg/g) M C D (μg/ml) V (ml) M (g) ㈡ 1. 1000 ± 0.002 g/l lanthan chloride heptahydrate (LaCl 3 7H 2 O) ICP-MSMerck 2. (flame atomic absorption spectrophotometer, FLAAS) Analyticjena ZEEnit 700 422.7 nm 3. AOAC 977.29 (11) 0.5 1.0 2.0 4.0 μg/ml 1~5 g (300 C)(600 C 4 )0.1 N HCl 100 ml0.5 ml LaCl 3 7H 2 O 0.13 g 50 ml 422.7 nm D ㈠ (mg/g) C x 200 x V 1 M 1000 C (μg/ml) V M (g) 35 32D 0.005 μg/g 18,000 μg/g 1 1800 9553 15D ㈡ 32 D17 ( 0.005 μg/g)15 D 0.042 5.13 μg/g D 176~8240% ( 29 33 644% 8240%) 80 120% 6 80% 2217 DD120% 4 1D0.005 μg/g0.4 μg/g 8240%3 80 120% 19%95
D 243 No. 29, 644 % No. 33, 8240 % No. 15, 1613 % No. 25, 1111 % D 29 33 644% 8240% 17%6 95 6 95 ㈠ 350.8 mg/g 128 mg/g ( ) 355 ㈡ 351.2 mg/g 154 mg/g 1.8 1613% (15 25 1613% 1111%)80 120% 9 80%18 120% 15 25 1613% 1111% 8 12 95 3 80 120%80% D 80% 80 120%120% ( )95 D 1. 2. 3. 2003 2003 () 2003[http://ceiba3. cc.ntu.edu.tw/course/4f786b/infof.htm]
244 Ann. Rept. BFDA 4. 5. 6. 7. 8. 9. 95 [http://food.doh. gov.tw/chinese/libary/libary2_3_9.htm] 2006 D 95 97 0971800323 D 2007 D 96 Heudi, O., Trisconi, M. J., and Blake, C. J. 2004. 95 97 D / 35 D/ / (%) ( ) D ( ) 80 18 (51) 22 (69) 80-120 9 (26) 6 (19) 120 8 (23) 4 (12) 35 32 Simultaneous quantification of vitamins A, D 3 and E in fortified infant formulae by liquid chromatography mass spectrometry. J Chromatogr. A. 1022 (1-2): 115-23. 10. Zoe, M., Dennis, J. W., and Sandra, J. R. 2005. Routine isotope-dilution liquid chromatography tandem mass spectrometry assay for simultaneous measurement of the 25-hydroxy metabolites of vitamins D 2 and D 3. Clin. Chem. 51(9): 1683-1690. 11. AOAC Official Methods of Analysis. 2000. A t o m i c a b s o r p t i o n s p e c t r o p h o t o m e t r i c flame photometric method. 18.4.09. AOAC International. U.S.A.
D 245 Survey on Compliance of Labeling of Vitamin D and Calcium on Dietary Supplements with Regulations in Taiwan CHING-HSIN TUNG, HUEI-WEN SHIAU, PIN-WEI TSENG, WEI-LUN TAI, JYH-QUAN PAN AND DANIEL YANG-CHIN SHIH Food Chemistry Divisçn ABSTRACT In this study, the dietary supplement samples were purchased from drugstores in Taipei, Taichung and Kaohsiung in 2008. On-line solid phase extract high performance liquid chromatography- ultraviolet (online SPE LC-UV) and On-line solid phase extract high performance liquid chromatography tandem mass spectrometry (online SPE LC\MS\MS) was employed to carry out the vitamin D analysis. Flame atomic absorption spectrophotometry (FLAAS) was applied to carry calcium assay. Thirty-two samples were analyzed for vitamin D, and the vitamin D contents of 6 samples detected fell within 80-120% of labeled values, 22 samples below 80%, and 4 samples over 120%. Thirty-five samples were analyzed for calcium. The calcium contents of 9 samples detected fell within 80-120% of labeled values, 18 samples below 80%, and 8 samples over 120%. This survey revealed that the labeled contents of vitamin D and calcium in marketed dietary supplements were all in compliance with the Regulation set by the Department of Health, but the conformation between assayed values and labeled values of vitamin D and calcium still needs to be improved. Key words: vitamin D 2 (ergocalciferol), vitamin D 3 (cholecalciferol), High performance liquid chromatography (HPLC), Flame atomic absorption spectrophotometry (FLAAS)