sign in sign up
<<
Home , Chlormadinone acetate, Diethylstilbestrol, Fluoxymesterone, Medroxyprogesterone, Methyltestosterone, Trenbolone

No. LCMSMS-065E

Liquid Chromatography Mass Spectrometry

Determination of 17 Residues in Milk by Ultra-High-Performance Liquid

Chromatography and Triple Quadrupole

No. LCMSMS-65E Mass Spectrometry

This application news presents a method for the determination of 17 hormone residues in milk using Shimadzu Ultra-High-Performance Liquid Chromatograph (UHPLC) LC-30A and Triple Quadrupole Mass Spectrometer LCMS- 8040. After sample pretreatment, the compounds in the milk matrix were separated using UPLC LC-30A and analyzed via Triple Quadrupole Mass Spectrometer LCMS-8040. All 17 displayed good linearity within their respective concentration range, with correlation coefficient in the range of 0.9974 and 0.9999. The RSD% of retention time and peak area of 17 hormones at the low-, mid- and high- concentrations were in the range of 0.0102-0.161% and 0.563-6.55% respectively, indicating good instrument precision. Method validation was conducted and the matrix spike recovery of milk ranged between 61.00-110.9%. The limit of quantitation was 0.14-0.975 g/kg, and it meets the requirement for detection of hormones in milk.

Keywords: Hormones; Milk; Solid phase extraction; Ultra performance liquid chromatograph; Triple quadrupole mass spectrometry

■ Introduction Since 2008’s -tainted milk scandal, the With reference to ’s national standard GB/T adulteration of milk powder has become a major 21981-2008 "Hormone Multi-Residue Detection safety concern. In recent years, another case of Method for Animal-derived Food - LC-MS Method", dairy product safety is suspected to cause "infant a method utilizing solid phase extraction, ultra- sexual precocity" (also known as ) performance liquid chromatography and triple and has become another major issue challenging the quadrupole mass spectrometry was developed for dairy industry in China. These frequent the rapid and high-sensitive detection of 17 crises on milk powder quality severely decrease hormones in milk. consumer confidence in domestic milk quality as well as impede the development of milk manufacturing ■ Experimental industry. Considering the interest and motivation of 1.1. Instruments manufacturers, it is apparent that hormones are not Shimadzu Ultra-High-Performance Liquid added during milk powder manufacturing and Chromatograph (UHPLC) Nexera X2 and Triple processing. However, farmers may use Quadrupole Mass Spectrometer LCMS-8040 system and hormones illegally on cows to increase the milk was used. The specific configuration included LC- yield for milk production. With the exception of 30AD×2 infusion pumps, DGU-20A5 Online endogenous , and Degasser, SIL-30AC Autosampler, CTO-30AC , all other chemically synthesized Column Oven, CBM-20A System Controller, Triple hormones in milk belong to prohibited drugs. In Quadrupole Mass Spectrometer LCMS-8040, and 2002, the Ministry of Agriculture of China issued an LabSolutions Ver. 5.53 Chromatography announcement (no. 235) prohibiting the use of Workstation. chemically synthesized hormones such as , , , and and stipulated that these compounds shall not be detected in any animal-derived food. At the same time, both EU directive 96/22/EC and US FDA have also banned the use of hormone drugs in animal- derived food. No. LCMSMS-065E

1.2. Analytical Conditions Liquid chromatography (LC) parameters Mass spectrometry (MS) conditions • Analyzer: Nexera X2 System • Analyzer: LCMS-8040 • Chromatographic column: Shimadzu Shim-pack XR- • source: ESI, positive ion scan ODS II 2.0 mm ID x 75 mmL, 2.2µm • Ion source interface voltage: 4.5 kV • Mobile phase: A-0.1% formic acid solution, B- • Nebulizing gas: Nitrogen 3.0 L/min methanol, A/B=50%/50% (V/V) • Drying gas: Nitrogen 15 L/min • Flow rate: 0.4 mL/min • Collision gas: Argon • Elution mode: gradient elution, refer to Table 1 • DL temperature: 250°C • volume: 5 μL • Heating module temperature: 400°C • Column temperature: 40°C • Scan mode: Multiple Reaction Monitoring (MRM) • Dwell time: 30 msec • Pause time: 3 msec • MRM parameters: Refer to Table 2 Table 1: Gradient Program Time(min) Module Command Value 8 Pumps PumpB Cone. 64 11 Pumps PumpB Cone. 64 12.5 Pumps PumpB Cone. 100 14.5 Pumps PumpB Cone. 100 15 Pumps PumpB Cone. 50 20 Controller Stop

Table 2: MRM Parameters Q1 Pre Bias Q3 Pre Bias Compound name Precursor ion Product ion CE (V) (V) (V) 287.05 97.00* -15.0 -25.0 -20.0 287.05 109.15 -15.0 -25.0 -23.0 287.10 121.05* -26.0 -25.0 -26.0 287.10 135.05 -26.0 -15.0 -15.0 338.25 148.15* -26.0 -30.0 -16.0 338.25 120.00 -26.0 -35.0 -24.0 337.15 241.15* -13.0 -25.0 -18.0 337.15 131.00 -13.0 -35.0 -28.0 289.25 97.00* -25.0 -25.0 -20.0 289.25 109.05 -25.0 -25.0 -22.0 303.25 109.15* -25.0 -30.0 -21.0 Methyltestosterone 303.25 97.15 -25.0 -25.0 -19.0 301.25 121.15* -26.0 -25.0 -13.0 Methandienone 301.25 149.15 -26.0 -15.0 -16.0 275.10 109.10* -13.0 -30.0 -22.0 275.10 257.20 -13.0 -15.0 -20.0 273.10 109.15* -10.0 -25.0 -23.0 Norandrostenedione 273.10 197.10 -10.0 -20.0 -15.0 271.20 253.15* -27.0 -20.0 -19.0 Trenbolone 271.20 199.05 -23.0 -25.0 -15.0 385.10 267.15* -23.0 -20.0 -20.0 acetate 385.10 325.20 -23.0 -15.0 -25.0 345.30 123.00* -13.0 -25.0 -13.0 Medroxyprogesterone 345.30 97.20 -13.0 -25.0 -19.0 387.30 123.00* -15.0 -30.0 -13.0 Medroxyprogesterone acetate 387.30 327.25 -15.0 -10.0 -25.0 313.25 109.15* -12.0 -35.0 -24.0 313.25 245.20 -12.0 -25.0 -18.0 405.05 345.20* -24.0 -15.0 -18.0 acetate 405.05 309.15 -24.0 -20.0 -23.0 299.05 109.05* -19.0 -35.0 -22.0 Norethindrone 299.05 91.15 -19.0 -25.0 -11.0 315.15 97.15* -12.0 -25.0 -20.0 315.15 109.00 -12.0 -30.0 -21.0 * Quantifier ion

No. LCMSMS-065E

1.3. Sample Preparation 1.3.1. Preparation of standard solutions subsequently diluted with water to prepare standard A mixed standard solution of 17 hormones, each working solutions at different concentration points containing 1000 µg/L, was prepared by diluting the as shown in Table 3. The standard calibration curves individual hormone standard (100 mg/L) with were established using these concentrations. ultrapure water. The mixed standard solution was

Table 3: Concentrations (μg/L) of 17 Hormones used for Calibration Compound name Conc. 1 Conc.2 Conc.3 Conc.4 Conc.5 Conc.6 Conc. 7 Norandrostenedione 1 2 4 10 20 50 100 Trenbolone 1 2 4 10 20 50 100 Boldenone 1 2 4 10 20 50 100 Nandrolone 1 2 4 10 20 50 100 Androstenedione 1 2 4 10 20 50 100 Methandienone 1 2 4 10 20 50 100 Testosterone 1 2 4 10 20 50 100 Methyltestosterone 1 2 4 10 20 50 100 Medroxyprogesterone 1 2 4 10 20 50 100 1 2 4 10 20 50 100 Medroxyprogesterone acetate 1 2 4 10 20 50 100 Progesterone 1 2 4 10 20 50 100 Norethindrone 5 10 20 50 100 250 500 Norgestrel 5 10 20 50 100 250 500 5 10 20 50 100 250 500 Fluoxymesterone 10 20 40 100 200 500 1000 Danazol 10 20 40 100 200 500 1000

1.3.2. Pretreatment of sample Milk samples were pretreated with reference to GB/T 21981-2008 "Hormone Multi-Residue Detection Method for Animal-derived Food - LC-MS Method".

■Results and discussion 2.1. Q1 Scan and Product Ion Scan Mass Spectra of the Hormone Standards

1. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -22V) (right) of androstenedione. No. LCMSMS-065E

2. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -21V) (right) of boldenone.

3. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -40V) (right) of danazol.

4. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -32V) (right) of fluoxymesterone.

No. LCMSMS-065E

5. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -25V) (right) of testosterone.

6. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -26V) (right) of methyltestosterone.

7. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -18V) (right) of methandienone.

No. LCMSMS-065E

8. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -24V) (right) of nandrolone.

9. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -25V) (right) of norandrostenedione.

10. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -30V) (right) of trenbolone.

No. LCMSMS-065E

11. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -19V) (right) of megestrol acetate.

12. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -29V) (right) of medroxyprogesterone.

13. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -20V) (right) of medroxyprogesterone acetate.

No. LCMSMS-065E

14. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -26V) (right) of norgestrel.

15. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -18V) (right) of chlormadinone acetate.

16. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -32V) (right) of norethindrone.

No. LCMSMS-065E

17. The Q1 scan mass spectrum (left) and Product Ion Scan mass spectrum (CE value was -25V) (right) of progesterone.

Figure 1: Q1 Scan and Product Ion Scan Mass Spectra of the Hormone Standards

2.2. MRM Chromatogram of the Mixed Hormones Standard Refer to Figure 2 for the MRM chromatogram of the mixed hormones standard.

Figure 2: MRM Chromatograms of the Hormone Standards Peak attribution: 1. Norandrostenedione (10 μg/L); 2. Trenbolone (10 μg/L); 3. Boldenone (10 μg/L); 4. Fluoxymesterone (100 μg/L); 5. Nandrolone (10 μg/L); 6. Androstenedione (10 μg/L); 7. Norethindrone (50 μg/L); 8. Methandienone (10 μg/L); 9. Testosterone (10 μg/L); 10. Norgestrel (50 μg/L); 11. Methyltestosterone (10 μg/L); 12. Medroxyprogesterone (10 μg/L); 13. Megestrol acetate (10 μg/L); 14. Chlormadinone acetate (50 μg/L); 15. Medroxyprogesterone acetate (10 μg/L); 16. Progesterone (10 μg/L); 17. Danazol (100 μg/L).

2.3. Calibration and Linearity Calibration was performed using a seven-point curve concentration range (Table 3) with the correlation and external standard method. Figure 3 shows the coefficient ranging between 0.9974 and 0.9999 calibration curves of all 17 hormones. Excellent (Table 4). linearity was obtained over the respective

No. LCMSMS-065E

1. Norandrostenedione 2. Trenbolone 3. Boldenone

4. Fluoxymesterone 5. Nandrolone 6. Androstenedione

7. Norethindrone 8. Methandienone 9. Testosterone

10. Norgestrel 11. Methyltestosterone 12. Medroxyprogesterone

No. LCMSMS-065E

13. Megestrol acetate 14. Chlormadinone acetate 15. Medroxyprogesterone acetate

16. Progesterone 17. Danazol

Figure 3: Calibration curves of all 17 hormones

Table 4: Calibration Curve Results of 17 Hormones No. Compound Name Calibration curve equation Linear range (μg/L) Correlation coefficient (R) 1 Norandrostenedione Y = (6291.94)X + (5045.17) 1.0-100 0.9998 2 Trenbolone Y = (8093.28)X + (11702.5) 1.0- 100 0.9999 3 Boldenone Y = (33290.1)X + (39696.4) 1.0- 100 0.9997 4 Fluoxymesterone Y = (1304.96)X + (21697.4) 10-1000 0.9991 5 Nandrolone Y = (10371.7)X + (11750.5) 1.0- 100 0.9997 6 Androstenedione Y= (9140.16)X + (4939.96) 1.0- 100 0.9999 7 Norethindrone Y = (3771.80)X + (44103.2) 5.0- 500 0.9993 8 Methandienone Y = (28151.5)X + (39506.0) 1.0- 100 0.9999 9 Testosterone Y = (17610.0)X + (13733.6) 1.0- 100 0.9999 10 Norgestrel Y = (3882.87)X + (56327.6) 5.0- 500 0.9992 11 Methyltestosterone Y = (14053.9)X + (28168.1) 1.0-100 0.9996 12 Medroxyprogesterone Y = (13545.6)X + (-8027.14) 1.0- 100 0.9985 13 Megestrol acetate Y = (17153.8)X+ (-18765.8) 1.0-100 0.9984 14 Chlormadinone acetate Y = (2524.61)X + (10654.8) 5.0- 500 0.9998 15 Medroxyprogesterone Y = (8715.12)X + (-11254 .1) 1.0- 100 0.9974 acetate 16 Progesterone Y= (22587.5)X + (-25767.7) 1.0- 100 0.9991 17 Danazol Y = (2422.08)X + (85881.9) 10- 1000 0.9977

2.4. Precision Experiment Mixed hormone standard solutions at low-, mid- and high- concentrations were prepared and analyzed (n=6). The RSD% obtained showed good instrument precision (Table 5).

No. LCMSMS-065E

Table 5: Repeatability Results of Retention Time and Peak Area (n=6) Sample name RSD% RSD% RSD% (low-conc, μg/L) (mid-conc, μg/L) (high-conc, μg/L) Norandrostenedione 0.1 1 5.82 0.115 2.32 0.0458 0.887 Trenbolone 0.125 4 83 0.0787 3.17 0.0400 1.12 Boldenone 0.0958 4.48 0.0767 2.60 0.0384 1.57 Fluoxymesterone 0.106 4.14 0.0700 2.40 0.0392 1.85 Nandrolone 0.131 6.55 0.0977 2.45 0.0444 0.715 Androstenedione 0.149 5.24 0.0333 1.71 0.0325 1.51 Norethindrone 0.123 0.909 0.0737 1.24 0.0364 0.745 Methandienone 0.0942 2.83 0.0635 1.88 0.0405 0.713 Testosterone 0.141 1.58 0.0578 3.53 0.0532 0.563 Norgestrel 0 .112 1.86 0.0414 2.14 0.0461 0.659 Methyltestosterone 0.160 2.27 0.0269 2.46 0.0472 1.54 Medroxyprogesterone 0.116 0.946 0.0147 3.65 0.0264 2.68 Megestrol acetate 0.120 5.25 0.0172 5.61 0.0271 2.78 Chlormadinone acetate 0 .0622 4.03 0.0196 5.27 0.0116 1.56 Medroxyprogesterone 0 .0862 6.54 0.0102 4.08 0.0169 2.51 acetate Progesterone 1.0658 4.99 0.0177 4.05 0.0138 2.04 Danazol 0.0566 5.09 0.0124 5.36 0.0168 2.74

1. Low concentration: The concentration of fluoxymesterone and danazol was 20 μg/L and the concentration of norethindrone, norgestrel and chlormadinone acetate was 10 μg/L, while the concentration of all the remaining hormones was 2 μg/L; 2. Mid-concentration: The concentration of fluoxymesterone and danazol was 100 μg/L and the concentration of norethindrone, norgestrel and chlormadinone acetate was 50 μg/L, while the concentration of all remaining hormones was 10 μg/L; 3. High concentration: The concentration of fluoxymesterone and danazol was 500 μg/L and the concentration of norethindrone, norgestrel and chlormadinone acetate was 250 μg/L, while the concentration of all remaining hormones was 50 μg/L.

2.5. Limit of Detection and Limit of Quantitation In order to examine the sensitivity of the instrument, based on the standard deviations (S) determined 7 standard hormone samples were prepared at from 7 injections. Method Detection Limit (MDL) is various concentrations (Table 6). Seven parallel calculated to be 3.14 x S, while the limit of sample injections were conducted, and the resulting quantitation (LOQ) is defined as 4 times of MDL. The chromatograms are shown in Figure 4. The limit of results are determined and shown in Table 6. detection and limit of quantitation were calculated

Table 6: Method Detection Limit and Limit of Quantitation of 17 Hormones No. Compound Name Concentration Standard Detection Limit of (μg/L) deviation (S) limit (μg/L) quantitation (μg/L) 1 Norandrostenedione 1 0.076 0.24 0.96 2 Trenbolone 1 0.062 0.19 0.76 3 Boldenone 1 0.060 0.19 0.76 4 Fluoxymesterone 10 0.069 0.22 0.87 5 Nandrolone 1 0.054 0.17 0.68 6 Androstenedione 1 0.089 0.28 1.12 7 Norethindrone 5 0.082 0.26 1.04 8 Methandienone 1 0.020 0.06 0.24 9 Testosterone 1 0.043 0.14 0.54 10 Norgestrel 5 0.11 0.34 1.36 11 Methyltestosterone 1 0.035 0.11 0.44 12 Medroxyprogesterone 1 0.060 0.19 0.76 13 Megestrol acetate 1 0.10 0.31 1.24 14 Chlormadinone acetate 5 0.071 0.22 0.88 15 Medroxyprogesterone acetate 1 0.079 0.25 1.00 16 Progesterone 1 0.059 0.19 0.76 17 Danazol 10 0.16 0.50 2.00

No. LCMSMS-065E

Figure 4: MRM Chromatograms of Hormone Standard Samples (Refer to Table 6 for the concentrations) Peak attribution: 1. Norandrostenedione; 2. Trenbolone; 3. Boldenone; 4. Fluoxymesterone; 5. Nandrolone; 6. Androstenedione; 7. Norethindrone; 8. Methandienone; 9. Testosterone; 10. Norgestrel; 11. Methyltestosterone; 12. Medroxyprogesterone; 13. Megestrol acetate; 14. Chlormadinone acetate; 15. Medroxyprogesterone acetate; 16. Progesterone; 17. Danazol.

2.6. Matrix Spike and Recovery Experiment Figure 5 shows the MRM chromatogram of the milk The results of detection and matrix spike recovery of sample prepared according to the method stated in these 17 hormones in milk are shown in Table 7. The 1.3. Based on the experimental results, progesterone limit of quantitation of this method was calculated to was detected. 17 hormone standards were spiked to be 0.14~0.97 μg/kg, where LOQ = 10 x S/N (Table the milk sample matrix, and the MRM chromatogram 8). after spiking are shown in Figure 6.

Figure 5: MRM Chromatogram of the Milk Matrix No. LCMSMS-065E

Figure 6: MRM Chromatogram of Spiked Milk Matrix (Refer to Table 7 for the concentrations and Figure 4 for peak attribution)

Table 7: Detection and Spiked Recovery Results of Hormones in Milk No. Compound Name Concentration of Spiking Concentration of Recovery hormones in concentration hormones in (%) non-spiked milk (μg/kg) spiked milk samples (μg/kg) samples (μg/kg) 1 Norandrostenedione N.D. 3.2 2.77 86.6 2 Trenbolone N.D. 3.2 2.39 74.7 3 Boldenone N.D. 3.2 2.59 80.9 4 Fluoxymesterone N.D. 32 27.1 84.7 5 Nandrolone N.D. 3.2 3.10 96.9 6 Androstenedione N.D. 3.2 3.00 93.8 7 Norethindrone N.D. 16 10.5 65.6 8 Methandienone N.D. 3.2 2.06 64.4 9 Testosterone N.D. 3.2 2.87 89.7 10 Norgestrel N.D. 16 10.9 68.1 11 Methyltestosterone N.D. 3.2 2.25 70.3 12 Medroxyprogesterone N.D. 3.2 3.55 110.9 13 Megestrol acetate N.D. 3.2 3.50 109.4 14 Chlormadinone acetate N.D. 16 14.5 90.6 15 Medroxyprogesterone acetate N.D. 3.2 3.30 103.1 16 Progesterone 1.5 3.2 4.74 101.2 17 Danazol N.D. 32 19.5 61.0 Note: N.D. represents not detected.

No. LCMSMS-065E

Table 8: Method Detection Limit and Limit of Quantitation

No. Compound Name Spiking Signal to noise Method Limit of concentration ratio (S/N) detection limit quantitation (μg/kg) (μg/kg) (μg/kg) 1 Norandrostenedione 1.6 48 0.10 0.33 2 Trenbolone 1.6 54 0.09 0.30 3 Boldenone 1.6 41 0.12 0.40 4 Fluoxymesterone 16 168 0.29 0.97 5 Nandrolone 1.6 55 0.09 0.30 6 Androstenedione 1.6 60 0.08 0.27 7 Norethindrone 8 140 0.17 0.57 8 Methandienone 1.6 89 0.05 0.17 9 Testosterone 1.6 116 0.04 0.14 10 Norgestrel 8 139 0.17 0.57 11 Methyltestosterone 1.6 96 0.05 0.17 12 Medroxyprogesterone 1.6 40 0.12 0.40 13 Megestrol acetate 1.6 54 0.09 0.30 14 Chlormadinone acetate 8 202 0.12 0.40 15 Medroxyprogesterone acetate 1.6 56 0.09 0.30 16 Progesterone 1.6 47 0.10 0.34 17 Danazol 16 171 0.28 0.93

■ Conclusion A method for the rapid determination of 17 The recovery of the spiked milk sample matrix hormone residues in milk was developed using ranged between 61.00% and 110.9%. The limit of Shimadzu Nexera X2 UHPLC and Triple Quadrupole quantitation of the method was determined to be Mass Spectrometer LCMS-8040. The linear range of 0.14~0.97 5g/kg; and the limit of detection is lower the calibration curves of these hormones was than the national standard stipulated in GB/T 21981- excellent with the correlation coefficients varying 2008 "Hormone Multi-Residue Detection Method for between 0.9974 and 0.9999. The precision was Animal-derived Food - LC-MS Method". The determined by analyzing consecutive 6 times (n = 6) described instrument and method demonstrates high for each of the low-, medium- and high- sensitivity and meets the requirements for detection concentration mixed standard solutions. The RSD of of hormones in milk. retention time and peak area obtained was 0.0102 - 0.161% and 0.563 - 6.55% respectively, indicating good instrument precision.

LCMS-8040 LCMS-8045 LCMS-8050 LCMS-8060 LCMS-2020 Q-TOF LCMS-9030

Founded in 1875, Shimadzu Corporation, a leader in the development of advanced technologies, has a distinguished history of innovation built on the foundation of contributing to society through science and technology. Established in 1975, Shimadzu Scientific Instruments (SSI), the American subsidiary of Shimadzu Corporation, provides a comprehensive range of analytical solutions to laboratories throughout North, Central, and parts of South America. SSI maintains a network of nine regional offices strategically located across the , with experienced technical specialists, service and sales engineers situated throughout the country, as well as applications laboratories on both coasts.

For information about Shimadzu Scientific Instruments and to contact your local office, please visit our Web site at www.ssi.shimadzu.com

Shimadzu Corporation For Research Use Only. Not for use in diagnostic procedures.The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu.The information contained herein is provided to you“as is” www.shimadzu.com/an/ without warranty of any kind including without limitation warranties as to is accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publications is SHIMADZU SCIENTIFIC INSTRUMENTS, INC. based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice. Applications Laboratory 7102 Riverwood Drive, Columbia, MD 21045 Phone:800-477-1227 Fax: 410-381-1222 © Shimadzu Scientific Instruments, 2018 URL http://www.ssi.shimadzu.com First Edition: July 2018