Nat Prod Sci.  2019 Mar;25(1):49-58. 10.20307/nps.2019.25.1.49.

Study on the Marker Steroids of New Zealand Deer (Cervus elaphus var. scoticus) Velvet Antler by UPLC-MS/MS and HPLC-PDA Methods - (II)

• Lee NK
• Jang KH
• Lee JT
• Park HW
• Han ST
• In G

• KMID: 2443104
• DOI: http://doi.org/10.20307/nps.2019.25.1.49

Abstract

Eleven steroid hormones (SHs: androstene-3,17-dione, estrone, β-estradiol, α-estradiol, testosterone, dehydroepiandrosterone, 17á-hydroxyprogesterone, medroxyprogesterone, megestrol acetate, progesterone, and androsterone) were detected from New Zealand deer (Cervus elaphus var. scoticus) velvet antler (NZA, 鹿茸). A method for the quantification of eleven SHs was established by using ultraperformance liquid chromatography (UPLC)-MS/MS. The linearities (R²> 0.991), limits of quantification (LOQ values, 0.3 ng/mL to 23.1 ng/mL), intraday and interday precisions (relative standard deviation: RSD < 2.43%), and recovery rates (97.3% to 104.6%) for all eleven SHs were determined. In addition, a method for the quantification of three 7-oxycholesterols (7-O-CSs: 7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol) in the NZA was established by using an HPLC-photodiode array (PDA) method. The linearities (R²> 0.999), LOQ values (30 ng/mL to 350 ng/mL), intraday and interday precisions (RSD < 1.93%), and recovery rates (97.2% to 103.5%) for the three 7-O-CSs were determined. These quantitative methods are accurate, precise, and reproducible. As a result, it is suggested that the five steroid compounds of androstene-3,17-dione, androsterone, 7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol could be marker steroids of NZA. These methods can be applied to quantify or standardize the marker steroids present in NZA.

Keyword

New Zealand deer antler; marker steroid; UPLC-MS/MS; HPLC-PDA; steroid hormone; 7-oxycholesterol

MeSH Terms

Androsterone
Animals
Antlers*
Chromatography, Liquid
Deer*
Dehydroepiandrosterone
Estrone
Medroxyprogesterone
Megestrol Acetate
Methods*
New Zealand*
Progesterone
Steroids*
Testosterone
Androsterone
Dehydroepiandrosterone
Estrone
Medroxyprogesterone
Megestrol Acetate
Progesterone
Steroids
Testosterone

Figure

• 

  Fig. 1. The total ion chromatograms for the eleven standard steroid hormones (1. androstene-3, 17-dione; 2. estrone; 3. β-estradiol; 4. α-estradiol; 5. testosterone; 6. dehydroepiandrosterone; 7. 17α-hydroxyprogesterone; 8. medroxyprogesterone; 9. megestrol acetate; 10. progesterone; 11. androsterone) (A) and New Zealand deer antler's 70% ethanol fraction (B) by UPLC-MS/MS in multiple reaction monitoring mode.

• 

  Fig. 2. The chromatograms for (A): the standard of three 7-oxycholesterols (7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol); (B): the 70% ethanol fraction (mixture of tip, upper, middle, and base) of New Zealand deer antler; (C): the chromatograph of 100 times the height of (B) by HPLC-PDA.

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