chp10AnalysisofSteroidHormoneDrugs

1、chapter 10 chapter 10 analysis of analysis of steroidsteroid hormone drugs hormone drugssection 1 gereral section 1 gereral considerations considerations 1. cyclopentanoperhydrophenanthrene 1. cyclopentanoperhydrophenanthrene skeleton skeleton abcd1234567891011121314151617 double bond double bond： d

2、ouble bond2. steroid hormones may be classified into: 2. steroid hormones may be classified into: adrenal cortex hormones (cortin) male hormones androgen & anabolic steroids female hormones progestogens (progestins) estrogens (esterins)56510142555(10)1,43 3adrenal cortex hormone:adrenal cortex hormo

3、ne:181920och3ch3oohhhhhhohcorticosteroneoch3ch3hohhohoohcortisonech3ch3hhohhhohoohohydrocortisoneooch3ch3hhhohoooch3prednisone aacetateoch3ch3hhohfhhch3ohooch3odexamethasone acetatech3ch3ohhhhffhooooch3ch3ooh3cfluocinonide211918character:character: 1) 21 c atomes 2) 4-3-one 3) c17- -ketol, or dihydr

4、oxyaceton side-chain 4) c11-carboxide, hydroxy group 5) others, such as: c1/c2- 1 6 , 9 -x 16 -oh 16 -ch34. androgens and anabolic steroids:4. androgens and anabolic steroids: the derivatives of testosterone the derivatives of testosterone(睾丸酮)ch3hch3ohhhhoch3hch3ohhhooch3testosteronetestosterone pr

5、opionatech3hch3ohhhch3omethyltestosteronehhch3hhhooonandrolone phenylpropionate character:character: 1) 19(18) c atomes 2) 4 -3-one 3) c17-oh(or ester), c17-ch3 4) c19-non-methyl (anabolic steroids)5. progestogensthe5. progestogensthe derivatives of progesterone derivatives of progesterone(黄体酮) ch3c

6、h3hhhhooch3progesteronech3ch3hhhooch3ooch3hydroxyprogesterone caproatech3hch3hch3hhoooch3och3medroxyprogesterone acetatech3hch3hhoooch3och3megestrol acetate character:character: 1) 21 c atomes 2) 4 -3-one 3) c17 methylketon, c17 -oh(ester) side-chain 4) others, such as: c6/c7- 6 6 -methyl6. estrogen

7、s6. estrogens derivatives of estradiolderivatives of estradiol(雌二醇)hch3ohhhhhoestradiolhch3ohhhhooestradiol benzoatehch3ohhhcchhoethinylestradiolhch3hhhooestronecharacter:character: 1) 18 c atomes 2) a ring : aromatic ring c3-oh (ph-oh), or esters 3) c17oh(or =o) and -cch(ethynyl) section 2 identifi

8、cationsection 2 identification 1.color-producing reaction: 1.color-producing reaction: 1) reaction of skeleton with strong acid color reagent: *sulfuric acid phosphoric acid perchloric acid hydrochloric acid 2) reaction of functional groups (1) c17- -ketol:ch3oohoxidantoch3oohohorch3oohoxidant: agno

9、3(nh3.h2o): ag fehling ts： cu2o tetrazolium(salt), such as: ttc(rt) or bt formazan (red or blue) (2) c17-methylketon: (progeaterone) sodium nitroferricyanideblue-violetch3och3och3ochno(cn)5fe 2-na fe(cn)5(no)(3) reaction with carbonyl reagents: c3,c20-one such as: phenylhydrazine sulfate isonicotiny

10、l hydrazine 2,4-dinitrophenylhydrazineco+h2nz-h2ocnzcondensionch3ohch3hhhch3och3ohnch3hhhch3nhonnonhnh2-h2o+(4) zimmermann reaction:coch2hohhch3ohhhoh-hohch3hhho-h2ono2o2nhohch3hhhoono2o2noh-hch3hhhoh2nnooo(5) c c3 3-oh (ph-oh) and c-oh (ph-oh) and c1717-cc:-cc:2. tlc2. tlc3. ir3. irch3hhhhohohagno3

11、hoch3hhhohccaghohhhch3ohcchso3-n+nhohch3hhnnho3shohsection 3 limit testsection 3 limit test1. test for foreign steroids1. test for foreign steroids：tlc, hplctlc, hplc2. test for free phosphate2. test for free phosphate： molybdenum blue colorimetrymolybdenum blue colorimetryh3po4ammonium molybdateh+(

12、nh4)3pmo12o40ammonium molybdophosphorate(nh4)2moo4oror5 (nh4)2o 12moo3 7h2oaammonium molybdophosphoratemolybdenum blueh(nh4)3p(mo3o10)4 n h2o3. test for free methanol and acetone3. test for free methanol and acetone： gcgc4. test for ethynyl4. test for ethynyl： agnoagno3 3 ts tssection 4 assaysection

13、 4 assay1. uv method:1. uv method: 1) structure: “c=c-c=c” or “c=c-c=o” (1) 4-3-one: max = 240nm (2) a ringaromatic ring: max = 280nm 2) discussion: (1) test for foreign steroids should be passed (2) be used for determination of material drug, not for preparations 2. tetrazolium colorimetry2. tetraz

14、olium colorimetry 1) principle : c17- -ketol may reduce the tetrazolium to form colored formazan (red or blue ) in strong alkaline solutionch3hhhohch3ooohohch3hhhch3ohoooh+ rt(bt)oh-+ formazan2) tetrazolium salts: red tetrazolium rt (ttc) blue tetrazolium bt 2e/2h+cl-nnnnnnhnn+ hclnnnnch3onnnnoch3cl

15、-cl-mono-formazanbis-formazan3) determination: uspbt bp, chprt4) discussion: (1) effects of substituents (2) effects of temperature and time of reaction: room temperature ttc: 1 hr bt: 90min (3) effects of oxygen in air and light: protected from light (4) effects of solvent and water : 95% alcohol,

16、water content 5% (5) effects of type and conc. of alkali: tetrabutylammonium hydroxide (6) effect of regents reaction sensitivity: bt ttc 3. inh colorimetry3. inh colorimetry 1) principle: c3-one or other carbonyl group may condense with carbonyl reagentsch3ohch3hhhch3och3ohnch3hhhch3nhonnonhnh2-h2o

17、+2) discussion: (1) selection of solvent: anhydrous ethanol (2) type and conc. of acid, and conc of inh hcl:inh2:1 (3) effects of water, temperature, light and oxygen in air (4) selectivity of reaction: 4-3-one(room temperature,1h, quantitative) c20-one c17-one c11-one (rt, 1h, not reaction) 4. kober reaction and4. kober reaction and iron-phenol reagent colorimetry iron-phenol reagent colorimetry 1) kober reaction : nmreddnmyellowethanolestrogens5154so2hor water 465-4so2hmaxmax2) iron-phenol reagent colorimetry: (1) iron-phenol reagent: (nh4)2fe(so4)26h2o + h2so4 + h2o2 nh4fe(so4