Simultaneous Determination Of Nine Forbidden Substances In Hair Growth Liquid By High Performance Liquid Chromatography-mass Spectrometry

Background:Alopecia is a common disease in Department of Dermatology,and the incidence of alopecia areata and seborrheic alopecia was the highest.With the development of the society,overwork,mental stress,irregular diet,air pollution,TV and computer radiation have become the major factors which lead people to alopecia.The alopecia population is increased quickly with the high growth rate of about 20%per year.It is helpful for promoting hair growth and reducing hair loss using the hair cosmetics,which is a kind of special cosmetics.Hair cosmetics have a very large market,and hair growth liquid is the solution type of hair care cosmetic.According to Hygienic Standard for Cosmetics which is edited in 2007 in China,some substances were banned in hair growth liquid,such as estriol,estradiol,estrone,diethylstilbestrol,testosterone,methyltestosterone,progesterone and minoxidil,and cantharidin is defined as a kind of restricted substance in hair growth liquid.The current standard methods can only detect few components with poor specificity and pollution,and some methods need a derivative process with complex operation.Therefore,it is necessary to explore an efficient and accurate method for the simultaneous determination of the substances with both qualitative and quantitative characteristics.Objective:To establish a method for the simultaneous determination of 9 restricted substances(cantharidin,minoxidil,estriol,estradiol,testosterone,methyltestosterone,estrone,diethylstilbestrol and progesterone)in hair growth liquid by using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Methods:The HPLC separation was performed on a CAPCELL PAK MG C18 column(2 mm×150 mm,5 ?m).The mobile phase was 10 mmol/L ammonium acetate solution(A)-acetonitrile(B)for gradient elution(0-12 min,90%A?50%A;12-13 min,50%A?60%A;13-20 min,60%A;20-30 min,60%A?20%A;30-31 min,20%A?90%A;31-40 min,90%A),the flow rate was 0.2 mL/min,and the injection volume was 5 ?L.Ion source was heat assisted electrospray ion source(HESI),and the ion source temperature was 100?.Positive ion and negative ion were used for detection,the ion source voltage was 4 kV in positive mode and 3 kV in negative mode.The capillary temperature was 350?,the sheath gas flow rate was 40 arb,and the auxiliary gas flow rate was 10 arb.With a scanning mode of selective reaction monitoring(SRM),the quasi-molecular ion of 9 restricted substances was selected as parent ion,two product ions with abundance stronger and less interference were selected as qualitative ion,and the fragment ion with the strongest intensity was selected as quantitative ion.Results:The established method had a strong specificity.There were a good linear correlation over the range of 1-100 ?g/mL for cantharidin,0.001-0.05 ?g/mL for minoxidil,0.25-25?g/mL for estriol,15-300 ?g/mL for estradiol,0.05-5 ?g/mL for testosterone,0.1-5 ?g/mL for methyltestosterone,0.06-3 ?g/mL for estrone,0.02-2 ?g/mL for diethylstilbestrol,0.05-5?g/mL for progesterone(r?0.998).The limits of detection of 9 restricted substances were 1.25?g/g,0.001 ?g/g,0.25 ?g/g,15 ?g/g,0.05 ?g/g,0.1 ?g/g,0.08 ?g/g,0.02 ?g/g and 0.05 ?g/g,respectively.The limit of quantification of 9 restricted substances were 5 ?g/g,0.005 ?g/g,1.25 ?g/g,75 ?g/g,0.25 ?g/g,0.5 ?g/g,0.3 ?g/g,0.1 ?g/g and 0.25 ?g/g,respectively.The recoveries of 9 restricted substances were between 80.5%-109.5%,and the RSDs were between 0.2%-5.1%.The established method had a high precision.Minoxidil was detected in two samples,and the contents were 1.7 mg/g and 2.8 mg/g respectively.Conclusion:The method can be used for simultaneous qualitative and quantitative determination of 9 restricted substances(7 sex hormones,cantharidin and minoxidil)in hair growth liquid.Specificity and sensitivity of the method are better than the existing detection methods.The method with strong anti-interference ability can be used for simultaneous qualitative and quantitative determination,and the method has a easy operating way.The method was suitable for simultaneous qualitative and quantitative determination of 9 restricted substances in hair growth liquid.The method is practical and provides scientific basis and technical support for cosmetics market supervision.