Summary
Synthetic steroid hormones have the ability to promote protein synthesis. The male-oriented action is the main features. It is also used in many aspects illegally. This post will detect the testosterone, methyltestosterone, progesterone, trenbolone and nearly 11 kinds of synthetic steroid hormones with High Performance Liquid Chromatography analysis-Tandem MS method.
Experimental part
Instruments and reagents:
TSQ Quantum triple quadrupole mass spectrometer equipped with ESI and Surveyor MS Pump LC System. Testosterone, Methyl testosterone, Progesterone, boldenone, Nandrolone, Methandienone, Stanozolol, Nandrolone propionate, Testosterone propionate and Nandrolone phenylpropionate. These reagents should be dissolved in methanol to 1.0mg / mL standard stock solution.
Sample preparation
Sampling
Weighing animal muscle and liver about 500g, chopping. Taking 10 eggs peeled and 500 ml fresh milk. Mixing them together.
Extracting
Taking 5g samples into a 50 mL centrifuge tubes, which contain 3 ml 10% sodium carbonate solution and 10 mL ethyl acetate homogenized 30s. Shaking 10 min and 6000 r / min centrifugation for 10 min. Transferring the upper organic phase into a pear shaped flask, and then it will be combined with ethyl acetate. After evaporated, it will be dissolved by 1 ml 50% aqueous acetonitrile. Taking proper amount of solution, which has been froze for 30 min and in 16000 r / min centrifugation for 5 min. And then it will be prepared for the LC/MS/MS detection.
LC-MS conditions
Column: Thermo Hypersil Gold (150 × 2.1 mm, 5 μm)
Electrospray ionization (ESI), positive ion mode, the spray voltage: 4500 V, the ion tube temperature: 350 ℃, selecting reaction monitoring (SRM) ion pair.
Conclusion
Sensitivity
This method is very sensitive in detecting 11 kinds of synthetic steroid hormones. As for pigs, cattle, sheep, chicken muscle tissue, eggs, testosterone, methyl testosterone, progesterone, trenbolone, boldenone, nandrolone, the United States androsterone, stanozolol, nandrolone propionate, testosterone propionate and nandrolone, the limit of the detection can reach at 0.1 μg / kg. And the Quantification limit is 0.5 μg / kg.
Accuracy
The extraction recovery of this method is 50% to 105%. And it is capable of accurate results and reproducible.
Synthetic steroid hormones have the ability to promote protein synthesis. The male-oriented action is the main features. It is also used in many aspects illegally. This post will detect the testosterone, methyltestosterone, progesterone, trenbolone and nearly 11 kinds of synthetic steroid hormones with High Performance Liquid Chromatography analysis-Tandem MS method.
Experimental part
Instruments and reagents:
TSQ Quantum triple quadrupole mass spectrometer equipped with ESI and Surveyor MS Pump LC System. Testosterone, Methyl testosterone, Progesterone, boldenone, Nandrolone, Methandienone, Stanozolol, Nandrolone propionate, Testosterone propionate and Nandrolone phenylpropionate. These reagents should be dissolved in methanol to 1.0mg / mL standard stock solution.
Sample preparation
Sampling
Weighing animal muscle and liver about 500g, chopping. Taking 10 eggs peeled and 500 ml fresh milk. Mixing them together.
Extracting
Taking 5g samples into a 50 mL centrifuge tubes, which contain 3 ml 10% sodium carbonate solution and 10 mL ethyl acetate homogenized 30s. Shaking 10 min and 6000 r / min centrifugation for 10 min. Transferring the upper organic phase into a pear shaped flask, and then it will be combined with ethyl acetate. After evaporated, it will be dissolved by 1 ml 50% aqueous acetonitrile. Taking proper amount of solution, which has been froze for 30 min and in 16000 r / min centrifugation for 5 min. And then it will be prepared for the LC/MS/MS detection.
LC-MS conditions
Column: Thermo Hypersil Gold (150 × 2.1 mm, 5 μm)
Electrospray ionization (ESI), positive ion mode, the spray voltage: 4500 V, the ion tube temperature: 350 ℃, selecting reaction monitoring (SRM) ion pair.
Conclusion
Sensitivity
This method is very sensitive in detecting 11 kinds of synthetic steroid hormones. As for pigs, cattle, sheep, chicken muscle tissue, eggs, testosterone, methyl testosterone, progesterone, trenbolone, boldenone, nandrolone, the United States androsterone, stanozolol, nandrolone propionate, testosterone propionate and nandrolone, the limit of the detection can reach at 0.1 μg / kg. And the Quantification limit is 0.5 μg / kg.
Accuracy
The extraction recovery of this method is 50% to 105%. And it is capable of accurate results and reproducible.