Oasis HLB Purification and ACQUITY UPLC/TQD quickly analyze the levels of glucosticoid stimulants in animal foods.

Summary:
This paper uses Waters® Oasis®HLB solid phase extraction technology and ACQUITY UPLC® ultra-efficient liquid phase
chromatography
/TQD series four poles
spectrometry
system to determine the analysis of glucosome
hormone
stimulants in animal foods.The chemical structure of

Glucocorticoids belongs to the steroid-like
compound
, which regulates sugar metabolism, promotes protein conversion to sugar, increases blood sugar concentration, anti-inflammatory, anti-allergic effects, and regulates water salt metabolism. Large intake of glucosal hormones from outside the body can cause intravoic hormone ratio disorders, sugar metabolism and inorganic salt metabolism disorders;
Long-term heavy use of glucodermal hormones can also induce or aggravate infections, cardiovascular system complications, osteoporosis, muscle atrophy, slow wound healing and digestive system complications, such as increased secretion of stomach acid, stomach
protease
, inhibit gastric mucus secretion, reduce resistance to gastrointestinal mucous membranes, and even cause bleeding or perforation of the digestive tract.
addition to having many side effects on human health, glucosal hormones can also affect the physical condition of athletes when they compete, contrary to the principle of fair competition in sports competitions, so such drugs belong to the International Olympic Committee announced the ban on anabolic steroids.
In recent years, health food, nutritional supplements and some animal foods for sports mobilization have been detected in a number of incidents containing glucosal hormones, due to the misuse of such contaminated foods, so that many athletes have unnecessary positive results.
2008 Beijing Olympic Games, in addition to strict doping testing of athletes, but also for athletes to eat all kinds of food for strict monitoring. In this paper, watershed HLB solid phase extraction purification technology and UPLC?/TQD system are used to establish fast, high-volume screening and corticosteroid screening and cortical hormones in animal foods, which provides the technical support of the whole program for stimulant monitoring of such foods.
Experimental Method
1.1
Reagents
and Materials
20-well vacuum extraction device, rotary evaporator, Oasis HLB (500mg, 6cc): ethyl acetate, methanol, foretic acid, ethyl acetate are pure chromatography, experimental water is ultra-pure water, sodium sulfate without water. Glucocorticoids
Sym standard
: pernisone, peronissone, hydrogenated pine, cortisone, methyl peronisone, python, tetrassemisson, perchromysone, fluorine-hydrogen pine, purity ≥98%.2.1 Analysis Conditions
Liquid Chromatography Conditions
LC System: Waters ACQUITY UPLCChromatography Column
: ACQUITY UPLC BEH C18, 2.1mm×5 0mm, 1.7m
column temperature: 30C
flow rate: 500 sl/min
flow phase A: water
flow phase B: acetylene
gradient: Time 0 min 20%B
time 5 min 25%B
time 5.3 min 50%B
time 5.5 min 60% B
time 5.6 min 20% B
6.5 min 20%B
Total analysis time 6.5 min
Samples: 10 μL
Mass Spectrometric Conditions
Mass Spectrometration System: Waters ACQUITY? TQD detector
ionized mode: ESI(-)
capillary voltage: 2.8 kV
solvent gas temperature: 400 degrees C
desolvable flow rate: 800 L/h
ion source temperature: Data acquisition
: Multi-reaction monitoring (MRM)
collision gas: argon 3.2×10-3 mba
-regulated ACQUITY TQD allows parent and child ions to have a resolution of 0.7 Da at half-peak width. The multi-reaction monitoring (MRM), resident time, tapered voltage, and collision energy list for this experiment can be found in Table 1. Underlined ion pairs are used for quantitative analysis.data acquisition and processing methods
waterse Mas Lyn x®4.1 operating software for data acquisition, TargetLynx® application management software for data processing.3.1 sample pre-treatment
said to take 5g of processed pork samples, in turn, added 10g waterless sodium sulfate, 20mL ethyl acetate, mixed uniform after homogeneity 1min, oscillation extraction 10min, centrifugation at 10000 rpm 5min. The liquid is transferred to the concentrated bottle, and then add 10mL ethyl acetate to the residue to repeat the above operation once, combine the two extracts, and rotate and evaporate to dry at 40 degrees C.
the sample was concentrated and dissolved in a 5mL 30% methanol aqueous solution. Oasis HLB is pre-active with 3mL methanol and 6mL water. All of the above extracts pass through the column, in turn with 5 mL of water, 5mL 50% methanol solution after washing and drying. With 10mL methanol / acetylene (1: 1) washed away and received, the sequestration at 40 degrees C nitrogen blow-dried, with acetylene / water (2: 8) dissolved to 1mL, after 0.2 m membrane on-machine analysis and determination..