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ELISA for the determination of saikosaponin a, an active component of Bupleuri Radix.
Chem Pharm Bull (Tokyo) 1998; 46(7):1140-3CP

Abstract

In order to quantify saikosaponin a (SSA), one of the major active components of Bupleuri Radix, a competitive and indirect ELISA method was developed. High titer rabbit polyclonal antibodies (pAbs) were raised against a conjugate of SSA and bovine serum albumin, coupled with a periodate oxidation method. SSA competitively inhibited the binding of rabbit anti-SSA pAbs to SSA-ovalbumin on the solid phase, a coated antigen on the well. The quantity of pAbs bound to the well was monitored using a peroxidase-conjugated anti-rabbit IgG as a secondary antibody, and tetramethylbenzidine solution as a substrate. The measuring range extended from 50 pg/ml to 20 ng/ml of SSA, with a detection limit of 40 pg/ml (5.13 pM). Antibodies showed some cross-reactivity with saikosaponin c (12.74%). However, the antibodies showed only slight cross-reactivities with saikosaponin d (0.3%), which differs from SSA only in the stereochemistry of the 16-hydroxyl group, and the artificial saikosaponins, saikosaponin b1 (2.1%) and saikosaponin g (0.53%). The specific and sensitive ELISA is especially suited for determination of SSA in samples when only small quantities of materials can be extracted for analysis.

Authors+Show Affiliations

College of Pharmacy, Chonnam National University, Kwangju, Korea.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

9692221

Citation

Jung, D W., et al. "ELISA for the Determination of Saikosaponin A, an Active Component of Bupleuri Radix." Chemical & Pharmaceutical Bulletin, vol. 46, no. 7, 1998, pp. 1140-3.
Jung DW, Shibuya M, Ebizuka Y, et al. ELISA for the determination of saikosaponin a, an active component of Bupleuri Radix. Chem Pharm Bull. 1998;46(7):1140-3.
Jung, D. W., Shibuya, M., Ebizuka, Y., Yoshimatsu, K., Shimomura, K., & Sung, C. K. (1998). ELISA for the determination of saikosaponin a, an active component of Bupleuri Radix. Chemical & Pharmaceutical Bulletin, 46(7), pp. 1140-3.
Jung DW, et al. ELISA for the Determination of Saikosaponin A, an Active Component of Bupleuri Radix. Chem Pharm Bull. 1998;46(7):1140-3. PubMed PMID: 9692221.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - ELISA for the determination of saikosaponin a, an active component of Bupleuri Radix. AU - Jung,D W, AU - Shibuya,M, AU - Ebizuka,Y, AU - Yoshimatsu,K, AU - Shimomura,K, AU - Sung,C K, PY - 1998/8/6/pubmed PY - 1998/8/6/medline PY - 1998/8/6/entrez SP - 1140 EP - 3 JF - Chemical & pharmaceutical bulletin JO - Chem. Pharm. Bull. VL - 46 IS - 7 N2 - In order to quantify saikosaponin a (SSA), one of the major active components of Bupleuri Radix, a competitive and indirect ELISA method was developed. High titer rabbit polyclonal antibodies (pAbs) were raised against a conjugate of SSA and bovine serum albumin, coupled with a periodate oxidation method. SSA competitively inhibited the binding of rabbit anti-SSA pAbs to SSA-ovalbumin on the solid phase, a coated antigen on the well. The quantity of pAbs bound to the well was monitored using a peroxidase-conjugated anti-rabbit IgG as a secondary antibody, and tetramethylbenzidine solution as a substrate. The measuring range extended from 50 pg/ml to 20 ng/ml of SSA, with a detection limit of 40 pg/ml (5.13 pM). Antibodies showed some cross-reactivity with saikosaponin c (12.74%). However, the antibodies showed only slight cross-reactivities with saikosaponin d (0.3%), which differs from SSA only in the stereochemistry of the 16-hydroxyl group, and the artificial saikosaponins, saikosaponin b1 (2.1%) and saikosaponin g (0.53%). The specific and sensitive ELISA is especially suited for determination of SSA in samples when only small quantities of materials can be extracted for analysis. SN - 0009-2363 UR - https://www.unboundmedicine.com/medline/citation/9692221/ELISA_for_the_determination_of_saikosaponin_a_an_active_component_of_Bupleuri_Radix_ DB - PRIME DP - Unbound Medicine ER -